LOCUS        pLV-EF1A-CD19.28H.28TM.28.3z       8753 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Modular CD28hinge-CD28transmembrane-CD28-CD3z CAR backbone, For 
            Transient Expression or Lentiviral Production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8753)
  AUTHORS   Timpanaro A, Piccand C, Dzhumashev D, Anton-Joseph S, Robbi A, Moser
            J, Rossler J, Bernasconi M
  TITLE     CD276-CAR T cells and Dual-CAR T cells targeting CD276/FGFR4 promote
            rhabdomyosarcoma clearance in orthotopic mouse models.
  JOURNAL   J Exp Clin Cancer Res. 2023 Nov 4;42(1):293. doi: 
            10.1186/s13046-023-02838-3.
  PUBMED    37924157
REFERENCE   2  (bases 1 to 8753)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            //gt12365.cn
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Exp Clin 
            Cancer Res."; date: "2023-11-4"; volume: "42(1):293. doi"; pages: " 
            10.1186/s13046-023-02838-3"
COMMENT     Sequence Label: pLV-EF1A-CD19.28H.28TM.28.3z
FEATURES             Location/Qualifiers
     source          1..8753
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1075..1308
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1803..1920
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    1938..1958
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        1965..2176
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     protein_bind    2182..2206
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2273..2302
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2303..2332
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3773..3829
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(3830..3832,3833..4546)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(3830..3832,3833..4543)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3872..3893)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4133..4152)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4480..4501
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(4556..4580)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4598..5186
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4651..4671)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5069..5080)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5274..5507
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5579..5700
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5616..5635)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5670..5689
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5740..5875
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5895..5914)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5896..5914)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5924..5941)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5924..5940)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5933..5955)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6082..6537
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6169..6188)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6379..6400
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6563..6667
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6668..6736,6737..7528)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6886..6905)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7699..8287
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8188..8207
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8441..8458
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8575..8596
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8611..8616,8617..8634,8635..8641)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8649..8665
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     8654..8676
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8673..8689
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8673..8689
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8708..8728
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8710..8728
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgcta
     1801 gcttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
     1861 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt
     1921 actagtgatt atcggatcaa ctttgtatag aaaagttgtc tagagggcag agcgcacatc
     1981 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
     2041 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     2101 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     2161 tgccgccaga acacagggat cccaagtttg tacaaaaaag caggctgcca ccatgctcag
     2221 gctgctcttg gctctcaact tattcccttc aattcaagta acaggactcg aggaacaaaa
     2281 actcatctca gaagaggatc tggaacaaaa gctcatttca gaagaagatc tgctcgaggt
     2341 cgacggagac gccgacatcc agatgacaca gactacatcc tccctgtctg cctctctggg
     2401 agacagagtc accatcagtt gcagggcaag tcaggacatc tctaagtatt tgaattggta
     2461 tcagcagaaa ccagatggaa ctgttaaact cctgatctac catacatcaa gattacactc
     2521 aggagtccca tcaaggttca gtggcagtgg gtctggaaca gattattctc tcaccattag
     2581 caacctggag caagaagata ttgccactta cttttgccaa cagggtaata cgcttccgta
     2641 cacgttcgga ggggggacta agttggaaat aacaggctcc acctctggat ccggcaagcc
     2701 cggatctggc gagggatcca ccaagggcga ggtgaaactg caggagtcag gacctggcct
     2761 ggtggcgccc tcacagagcc tgtccgtcac atgcactgtc tcaggggtct cattacccga
     2821 ctatggtgta agctggattc gccagcctcc acgaaagggt ctggagtggc tgggagtaat
     2881 atggggtagt gaaaccacat actataattc agctctcaaa tccagactga ccatcatcaa
     2941 ggacaactcc aagagccaag ttttcttaaa aatgaacagt ctgcaaactg atgacacagc
     3001 catttactac tgtgccaaac attattacta cggtggtagc tatgctatgg actactgggg
     3061 tcaaggaacc tcagtcaccg tctccggtac cattgaagtt atgtatcctc ctccttacct
     3121 agacaatgag aagagcaatg gaaccattat ccatgtgaaa gggaaacacc tttgtccaag
     3181 tcccctattt cccggacctt ctaagccctt ttgggtgctg gtggtggttg gtggagtcct
     3241 ggcttgctat agcttgctag taacagtggc ctttattatt ttctgggtga ggagtaagag
     3301 gagcaggctc ctgcacagtg actacatgaa catgactccc aggcggcccg gacccacccg
     3361 caagcattac cagccctatg ccccaccacg cgacttcgca gcctatcgct cctccggact
     3421 gagagtgaag ttcagcagga gcgcagacgc ccccgcgtac cagcagggcc agaaccagct
     3481 ctataacgag ctcaatctag gacgaagaga ggagtacgat gttttggaca agaggcgtgg
     3541 ccgggaccct gagatggggg gaaagccgca gagaaggaag aaccctcagg aaggcctgta
     3601 caatgaactg cagaaagata agatggcgga ggcctacagt gagattggga tgaaaggcga
     3661 gcgccggagg ggcaaggggc acgatggcct ttaccagggt ctcagtacag ccaccaagga
     3721 cacctacgac gcccttcaca tgcaggccct gccccctcgc tccggacgta cggccacgaa
     3781 cttctctctg ttaaagcaag caggcgacgt ggaagaaaac cccggtcccg tgagcaaggg
     3841 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     3901 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     3961 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     4021 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     4081 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     4141 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     4201 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     4261 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     4321 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     4381 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     4441 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     4501 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagggc cccgtaccca
     4561 gctttcttgt acaaagtggt gataatcgaa ttccgataat caacctctgg attacaaaat
     4621 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     4681 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     4741 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     4801 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     4861 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     4921 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     4981 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     5041 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     5101 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     5161 gatctccctt tgggccgcct ccccgcatcg ggaattcccg cggttcgctt taagaccaat
     5221 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     5281 gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt
     5341 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     5401 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     5461 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     5521 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
     5581 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5641 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5701 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc
     5761 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5821 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg
     5881 acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
     5941 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     6001 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     6061 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     6121 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     6181 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     6241 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     6301 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     6361 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     6421 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     6481 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     6541 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6601 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6661 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6721 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6781 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6841 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6901 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6961 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     7021 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     7081 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     7141 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     7201 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     7261 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     7321 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     7381 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     7441 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     7501 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     7561 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     7621 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7681 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7741 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7801 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     7861 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7921 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7981 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     8041 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     8101 cgccacgctt cccgaagaga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     8161 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     8221 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     8281 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     8341 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     8401 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     8461 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     8521 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     8581 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     8641 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     8701 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag ctt
//