LOCUS        pHAGE-CMV-hIL7-IRES-ZsGreen-W                8250 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pHAGE-CMV-hIL7-IRES-ZsGreen-W
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8250)
  TITLE     David Baltimore's unpublished plasmids
REFERENCE   2  (bases 1 to 8250)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            //gt12365.cn
FEATURES             Location/Qualifiers
     source          1..8250
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2227..2530
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2531..2734
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2685..2705
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2731..2755
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    3350..3917
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3367..3919
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3511..3528)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3738..3757
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3931..4626
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     misc_feature    4640..5228
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4693..4713)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5111..5122)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5303..5536
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5655..5673)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5741..5845
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5846..6706
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6064..6083)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6877..7465
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7366..7385
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7619..7636
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7728..7747
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     protein_bind    7876..7897
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7912..7942
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7950..7966
                     /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     7955..7977
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7974..7990
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7974..7990
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8040..8059
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtgga
     2221 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     2281 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     2341 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     2401 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     2461 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     2521 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     2581 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     2641 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     2701 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg
     2761 gagacgccat ccacgctgtt ttgacctcca tagaagacac cggcggccgc catgttccat
     2821 gtttctttta ggtatatctt tggacttcct cccctgatcc ttgttctgtt gccagtagca
     2881 tcatctgatt gtgatattga aggtaaagat ggcaaacaat atgagagtgt tctaatggtc
     2941 agcatcgatc aattattgga cagcatgaaa gaaattggta gcaattgcct gaataatgaa
     3001 tttaactttt ttaaaagaca tatctgtgat gctaataagg aaggtatgtt tttattccgt
     3061 gctgctcgca agttgaggca atttcttaaa atgaatagca ctggtgattt tgatctccac
     3121 ttattaaaag tttcagaagg cacaacaata ctgttgaact gcactggcca ggttaaagga
     3181 agaaaaccag ctgccctggg tgaagcccaa ccaacaaaga gtttggaaga aaataaatct
     3241 ttaaaggaac agaaaaaact gaatgacttg tgtttcctaa agagactatt acaagagata
     3301 aaaacttgtt ggaataaaat tttgatgggc actaaagaac actaaaggat ccctcccccc
     3361 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     3421 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
     3481 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
     3541 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
     3601 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
     3661 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
     3721 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
     3781 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
     3841 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     3901 gaaaaacacg atgataatat ggccacacat atggcccagt ccaagcacgg cctgaccaag
     3961 gagatgacca tgaagtaccg catggagggc tgcgtggacg gccacaagtt cgtgatcacc
     4021 ggcgagggca tcggctaccc cttcaagggc aagcaggcca tcaacctgtg cgtggtggag
     4081 ggcggcccct tgcccttcgc cgaggacatc ttgtccgccg ccttcatgta cggcaaccgc
     4141 gtgttcaccg agtaccccca ggacatcgtc gactacttca agaactcctg ccccgccggc
     4201 tacacctggg accgctcctt cctgttcgag gacggcgccg tgtgcatctg caacgccgac
     4261 atcaccgtga gcgtggagga gaactgcatg taccacgagt ccaagttcta cggcgtgaac
     4321 ttccccgccg acggccccgt gatgaagaag atgaccgaca actgggagcc ctcctgcgag
     4381 aagatcatcc ccgtgcccaa gcagggcatc ttgaagggcg acgtgagcat gtacctgctg
     4441 ctgaaggacg gtggccgctt gcgctgccag ttcgacaccg tgtacaaggc caagtccgtg
     4501 ccccgcaaga tgcccgactg gcacttcatc cagcacaagc tgacccgcga ggaccgcagc
     4561 gacgccaaga accagaagtg gcacctgacc gagcacgcca tcgcctccgg ctccgccttg
     4621 ccctgaatcg atagatccta atcaacctct ggattacaaa atttgtgaaa gattgactgg
     4681 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     4741 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     4801 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     4861 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     4921 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     4981 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     5041 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     5101 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     5161 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     5221 ctccccgcct gagatccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     5281 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
     5341 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     5401 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     5461 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     5521 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     5581 aagaaatgaa tatcagagag tgagaggccc gggttaatta aggaaagggc tagatcattc
     5641 ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     5701 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5761 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5821 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5881 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5941 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     6001 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     6061 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     6121 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6181 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6241 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6301 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6361 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     6421 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6481 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6541 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6601 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6661 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6721 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6781 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6841 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6901 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6961 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     7021 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     7081 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7141 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7201 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     7261 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7321 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     7381 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7441 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7501 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     7561 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     7621 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     7681 ttggccgatt cattaatgca gcaagctcat ggctgactaa ttttttttat ttatgcagag
     7741 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     7801 ctaggctttt gcaaaaagct ccccgtggca cgacaggttt cccgactgga aagcgggcag
     7861 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     7921 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     7981 cagctatgac atgattacga atttcacaaa taaagcattt ttttcactgc attctagttg
     8041 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc
     8101 taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg
     8161 gtttcattta ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat
     8221 ttgttaaata tgtactacaa acttagtagt
//