LOCUS       pHyo245       10178 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses pheS_A294G by dual T7 promoter in e.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10178)
  AUTHORS   Park H, Kim S
  TITLE     Gene-specific mutagenesis enables rapid continuous evolution of 
            enzymes in vivo.
  JOURNAL   Nucleic Acids Res. 2021 Apr 6;49(6):e32. doi: 10.1093/nar/gkaa1231.
  PUBMED    33406230
REFERENCE   2  (bases 1 to 10178)
  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: "Nucleic 
            Acids Res."; date: "2021-04-6"; volume: "49(6):e32. doi"; pages: " 
            10.1093/nar/gkaa1231"
COMMENT     Sequence Label: pHyo245
FEATURES             Location/Qualifiers
     source          1..10178
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(820..867)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     863..881
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     promoter        915..933
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(966..983)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1977..1994)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(2004..2021)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             2040..2062
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2077..2101
                     /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)."
     promoter        complement(2102..2120)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2198..2245
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2252..2271)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2477..2554
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2555..3637
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(2574..2593)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             complement(join(3954..4745,4746..4814))
                     /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     4577..4596
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4815..4919)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5543..6178
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      6265..6484
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             6575..7330
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    7333..7583
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             7909..9084
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             9084..10055
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    join(10128..10178,1..423)
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
ORIGIN
        1 taatcagact gggaccacgg tcccactcgt atcgtcggtc tgattattag tctgggacca
       61 cggtcccact cgtatcgtcg gtctgattat tagtctggga ccacggtccc actcgtatcg
      121 tcggtctgat aatcagactg ggaccacggt cccactcgta tcgtcggtct gattattagt
      181 ctgggaccat ggtcccactc gtatcgtcgg tctgattatt agtctgggac cacggtccca
      241 ctcgtatcgt cggtctgatt attagtctgg aaccacggtc ccactcgtat cgtcggtctg
      301 attattagtc tgggaccacg gtcccactcg tatcgtcggt ctgattatta gtctgggacc
      361 acgatcccac tcgtgttgtc ggtctgatta tcggtctggg accacggtcc cacttgtatt
      421 gtcgatcaga ctatcagcgt gagactacga ttccatcaat gcctgtcaag ggcaagtatt
      481 gacatgtcgt cgtaacctgt agaacggagt aacctcggtg tgcggttgta tgcctgctgt
      541 ggattgctgc tgtgtcctgc ttatccacaa cattttgcgc acggttatgt ggacaaaata
      601 cctggttacc caggccgtgc cggcacgtta accgggctgc atccgatgca agtgtgtcgc
      661 tgtcgagtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcccgtacg
      721 gccgactagt aggcctatta tttttgacac cagacaagtt ggtaatggta gcgaccggcg
      781 ctcagttgga attcatccgg atatagttcc tcctttcagc aaaaaacccc tcaagacccg
      841 tttagaggcc ccaaggggtt atgctagtta ttgctcagcg gtggcagcag ccaactcagc
      901 ttcctttccg aaattaatac gactcactat aggggaattg tggctttgtt agcagccgga
      961 tctcagtggt ggtggtggtg gtgctcgagt tatttaaact gtttgaggaa acgcagatcg
     1021 ttttcgaaga atgaacgcag gtcggtgacg ccgtaacgca acatagtcag acgctccatc
     1081 cccatcccga agccgaaacc agagtaaact tccgggtcga tgccaacgtt acgcaacacg
     1141 ttcggatgca ccatcccgca gcccagcact tccagccatt taccgttttt acccatgacg
     1201 tccacttccg cagaaggttc ggtaaacggg aagtaggaag gacggaagcg aatctgcaaa
     1261 tcttcctcaa agaagttacg caggaagtcg tgcagcgtgc ctttcaggtt ggtaaagctg
     1321 atgttggtat caacaatcag accttccatc tgatggaaca tcggcgtgtg agtctggtcg
     1381 tagtcgttac gataaacacg gccaggcgcg atgatacgaa tcggtggctg ctgggctttc
     1441 atggtgcgga tctgtacgcc agaggtctgg gtacgcagca ggcgggtagt gtcaaaccag
     1501 aaagtgtcgt ggtcagcgcg cgccgggtgg tgaccaggaa tgttcagagc atcgaagtta
     1561 tgataatcgt cttcgatttc cggcccggtt gccacggtaa agccaagctc accgaagaaa
     1621 ctttcgatac ggtcgatggt acgggtaacc ggatgcagac cgccgttttc aatgcgacga
     1681 cctggcagag agacatcaat cgtttccgcc gccagacgcg cattcagtgc agcgctttcc
     1741 agttccgctt tacgcgcatt cagcgcctgc tgaacctgct ctttcgcttc gttgataacc
     1801 gcaccagctg ccggacgctc ttctggcggc agctcacgca gggtcgtcat ctgaagggtt
     1861 aagtgccctt ttttacccaa atattcgacg cgcacattat ctaacgcggc aacatctgac
     1921 gcctggctaa tggccgcctt cgcactggca accagttctg cgagatgtga catatggctg
     1981 ccgcgcggca ccaggccgct gctgtgatga tgatgatgat ggctgctgcc catggtatat
     2041 ctccttctta aagttaaaca aaattatttc tagaggggaa ttgttatccg ctcacaattc
     2101 ccctatagtg agtcgtatta atttcgcggg atcgagatct cgatcctcta cgccggacgc
     2161 atcgtggccg gcatcaccgg cgccacaggt gcggttgcta gcataacccc ttggggcctc
     2221 taaacgggtc ttgaggggtt ttttgctggc gcctatatcg ccgacatcac cgatggggaa
     2281 gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc
     2341 cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct tgcggcggcg
     2401 gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga
     2461 gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata
     2521 gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt
     2581 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
     2641 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
     2701 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
     2761 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
     2821 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc
     2881 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
     2941 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
     3001 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact
     3061 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
     3121 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
     3181 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
     3241 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
     3301 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc
     3361 ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat
     3421 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
     3481 gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac
     3541 cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     3601 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt
     3661 agctcactca ttaggcaccg ggatctcgac cgatgccctt gagagccttc aacccagtca
     3721 gctccttccg gtggaagcag cataaccttt ttccgtgatg gtaacttcac ggtaaccaag
     3781 atgctcgacc aattctcatg tttgacagct tatcatcgaa tttctgccat tcatccgctt
     3841 attatcactt attcaggcgt agcaaccagg cgtttaaggg caccaataac tgccttaaaa
     3901 aaattacgcc ccgccctgcc actcatcgca gttgagtaaa cttggtctga cagttaccaa
     3961 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4021 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4081 gcaatgatac cgcgagaacc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4141 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4201 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4261 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4321 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4381 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4441 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4501 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4561 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4621 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4681 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4741 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4801 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4861 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggtttaccgc
     4921 tgaaagttct gcaaagcctg atgggacata agtccatcag ttcaacggaa gtctacacga
     4981 aggtttttgc gctggatgtg gctgcccggc accgggtgca gtttgcgatg ccggagtctg
     5041 atgcggttgc gatgctgaaa caattatcct gagaataaat gccttggcct ttatatggaa
     5101 atgtggaact gagtggatat gctgtttttg tctgttaaac agagaagctg gctgttatcc
     5161 actgagaagc gaacgaaaca gtcgggaaaa tctcccatta tcgtagagat ccgcattatt
     5221 aatctcagga gcctgtgtag cgtttatagg aagtagtgtt ctgtcatgat gcctgcaagc
     5281 ggtaacgaaa acgatttgaa tatgccttca ggaacaatag aaatcttcgt gcggtgttac
     5341 gttgaagtgg agcggattat gtcagcaatg gacagaacaa cctaatgaac acagaaccat
     5401 gatgtggtct gtccttttac agccagtagt gctcgccgca gtcgagcgac agggcgaagc
     5461 cctcgagatc ccctgccatt tttggggtga ggccgttcgc ggccgagggg cgcagcccct
     5521 ggggggatgg gaggcccgcg ttagcgggcc gggagggttc gagaaggggg ggcacccccc
     5581 ttcggcgtgc gcggtcacgc gcacagggcg cagccctggt taaaaacaag gtttataaat
     5641 attggtttaa aagcaggtta aaagacaggt tagcggtggc cgaaaaacgg gcggaaaccc
     5701 ttgcaaatgc tggattttct gcctgtggac agcccctcaa atgtcaatag gtgcgcccct
     5761 catctgtcag cactctgccc ctcaagtgtc aaggatcgcg cccctcatct gtcagtagtc
     5821 gcgcccctca agtgtcaata ccgcagggca cttatcccca ggcttgtcca catcatctgt
     5881 gggaaactcg cgtaaaatca ggcgttttcg ccgatttgcg aggctggcca gctccacgtc
     5941 gccggccgaa atcgagcctg cccctcatct gtcaacgccg cgccgggtga gtcggcccct
     6001 caagtgtcaa cgtccgcccc tcatctgtca gtgagggcca agttttccgc gaggtatcca
     6061 caacgccggc ggccggccgc ggtgtctcgc acacggcttc gacggcgttt ctggcgcgtt
     6121 tgcagggcca tagacggccg ccagcccagc ggcgagggca accagcccgg tgagcgtcgg
     6181 aaaggggaat ttcgagtgag cgaggaagca ccagggaaca gcacttatat attctgctta
     6241 cacacgatgc ctgaaaaaac ttcccttggg gttatccact tatccacggg gatattttta
     6301 taattatttt ttttatagtt tttagatctt cttttttaga gcgccttgta ggcctttatc
     6361 catgctggtt ctagagaagg tgttgtgaca aattgccctt tcagtgtgac aaatcaccct
     6421 caaatgacag tcctgtctgt gacaaattgc ccttaaccct gtgacaaatt gccctcagaa
     6481 gaagctgttt tttcacaaag ttatccctgc ttattgactc ttttttattt agtgtgacaa
     6541 tctaaaaact tgtcacactt cacatggatc tgtcatggcg gaaacagcgg ttatcaatca
     6601 caagaaacgt aaaaatagcc cgcgaatcgt ccagtcaaac gacctcactg aggcggcata
     6661 tagtctctcc cgggatcaaa aacgtatgct gtatctgttc gttgaccaga tcagaaaatc
     6721 tgatggcacc ctacaggaac atgacggtat ctgcgagatc catgttgcta aatatgctga
     6781 aatattcgga ttgacctctg cggaagccag taaggatata cggcaggcat tgaagagttt
     6841 cgcggggaag gaagtggttt tttatcgccc tgaagaggat gccggcgatg aaaaaggcta
     6901 tgaatctttt ccttggttta tcaaacgtgc gcacagtcca tccagagggc tttacagtgt
     6961 acatatcaac ccatatctca ttcccttctt tatcgggtta cagaaccggt ttacgcagtt
     7021 tcggcttagt gaaacaaaag aaatcaccaa tccgtatgcc atgcgtttat acgaatccct
     7081 gtgtcagtat cgtaagccgg atggctcagg catcgtctct ctgaaaatcg actggatcat
     7141 agagcgttac cagctgcctc aaagttacca gcgtatgcct gacttccgcc gccgcttcct
     7201 gcaggtctgt gttaatgaga tcaacagcag aactccaatg cgcctctcat acattgagaa
     7261 aaagaaaggc cgccagacga ctcatatcgt attttccttc cgcgatatca cttccatgac
     7321 gacaggatag tctgagggtt atctgtcaca gatttgaggg tggttcgtca catttgttct
     7381 gacctactga gggtaatttg tcacagtttt gctgtttcct tcagcctgca tggattttct
     7441 catacttttt gaactgtaat ttttaaggaa gccaaatttg agggcagttt gtcacagttg
     7501 atttccttct ctttcccttc gtcatgtgac ctgatatcgg gggttagttc gtcatcattg
     7561 atgagggttg attatcacag tttattactc tgaattggct atccgcgtgt gtacctctac
     7621 ctggagtttt tcccacggtg gatatttctt cttgcgctga gcgtaagagc tatctgacag
     7681 aacagttctt ctttgcttcc tcgccagttc gctcgctatg ctcggttaca cggctgcggc
     7741 gagcgctagt gataataagt gactgaggta tgtgctcttc ttatctcctt ttgtagtgtt
     7801 gctcttattt taaacaactt tgcggttttt tgatgacttt gcgattttgt tgttgctttg
     7861 cagtaaattg caagatttaa taaaaaaacg caaagcaatg attaaaggat gttcagaatg
     7921 aaactcatgg aaacacttaa ccagtgcata aacgctggtc atgaaatgac gaaggctatc
     7981 gccattgcac agtttaatga tgacagcccg gaagcgagga aaataacccg gcgctggaga
     8041 ataggtgaag cagcggattt agttggggtt tcttctcagg ctatcagaga tgccgagaaa
     8101 gcagggcgac taccgcaccc ggatatggaa attcgaggac gggttgagca acgtgttggt
     8161 tatacaattg aacaaattaa tcatatgcgt gatgtgtttg gtacgcgatt gcgacgtgct
     8221 gaagacgtat ttccaccggt gatcggggtt gctgcccata aaggtggcgt ttacaaaacc
     8281 tcagtttctg ttcatcttgc tcaggatctg gctctgaagg ggctacgtgt tttgctcgtg
     8341 gaaggtaacg acccccaggg aacagcctca atgtatcacg gatgggtacc agatcttcat
     8401 attcatgcag aagacactct cctgcctttc tatcttgggg aaaaggacga tgtcacttat
     8461 gcaataaagc ccacttgctg gccggggctt gacattattc cttcctgtct ggctctgcac
     8521 cgtattgaaa ctgagttaat gggcaaattt gatgaaggta aactgcccac cgatccacac
     8581 ctgatgctcc gactggccat tgaaactgtt gctcatgact atgatgtcat agttattgac
     8641 agcgcgccta acctgggtat cggcacgatt aatgtcgtat gtgctgctga tgtgctgatt
     8701 gttcccacgc ctgctgagtt gtttgactac acctccgcac tgcagttttt cgatatgctt
     8761 cgtgatctgc tcaagaacgt tgatcttaaa gggttcgagc ctgatgtacg tattttgctt
     8821 accaaataca gcaatagtaa tggctctcag tccccgtgga tggaggagca aattcgggat
     8881 gcctggggaa gcatggttct aaaaaatgtt gtacgtgaaa cggatgaagt tggtaaaggt
     8941 cagatccgga tgagaactgt ttttgaacag gccattgatc aacgctcttc aactggtgcc
     9001 tggagaaatg ctctttctat ttgggaacct gtctgcaatg aaattttcga tcgtctgatt
     9061 aaaccacgct gggagattag ataatgaagc gtgcgcctgt tattccaaaa catacgctca
     9121 atactcaacc ggttgaagat acttcgttat cgacaccagc tgccccgatg gtggattcgt
     9181 taattgcgcg cgtaggagta atggctcgcg gtaatgccat tactttgcct gtatgtggtc
     9241 gggatgtgaa gtttactctt gaagtgctcc ggggtgatag tgttgagaag acctctcggg
     9301 tatggtcagg taatgaacgt gaccaggagc tgcttactga ggacgcactg gatgatctca
     9361 tcccttcttt tctactgact ggtcaacaga caccggcgtt cggtcgaaga gtatctggtg
     9421 tcatagaaat tgccgatggg agtcgccgtc gtaaagctgc tgcacttacc gaaagtgatt
     9481 atcgtgttct ggttggcgag ctggatgatg agcagatggc tgcattatcc agattgggta
     9541 acgattatcg cccaacaagt gcttatgaac gtggtcagcg ttatgcaagc cgattgcaga
     9601 atgaatttgc tggaaatatt tctgcgctgg ctgatgcgga aaatatttca cgtaagatta
     9661 ttacccgctg tatcaacacc gccaaattgc ctaaatcagt tgttgctctt ttttctcacc
     9721 ccggtgaact atctgcccgg tcaggtgatg cacttcaaaa agcctttaca gataaagagg
     9781 aattacttaa gcagcaggca tctaaccttc atgagcagaa aaaagctggg gtgatatttg
     9841 aagctgaaga agttatcact cttttaactt ctgtgcttaa aacgtcatcc gcatcaagaa
     9901 ctagtttaag ctcacgacat cagtttgctc ctggagcgac agtattgtat aagggcgata
     9961 aaatggtgct taacctggac aggtctcgtg ttccaactga gtgtatagag aaaattgagg
    10021 ccattcttaa ggaacttgaa aagccagcac cctgatgcga ccacgtttta gtctacgttt
    10081 atctgtcttt acttaatgtc ctttgttaca ggccagaaag cataactggc ctgaatattc
    10141 tctctgggcc cactgttcca cttgtatcgt cggtctga
//