LOCUS       V025894                10358 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025894
VERSION     V025894
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     promoter        2337..3519
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     CDS             3547..5076
                     /label="NECTIN4(NM_030916)"
                     /note="NECTIN4(NM_030916)"
                     /gene="NECTIN4"
     misc_feature    5108..5694
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             5714..6313
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    6327..6915
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             7122..7755
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7883..7899)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7907..7923
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(7931..7961)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7976..7997
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8285..8873)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9044..9904)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(9905..10009)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    10057..10191
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
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 tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg
     2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc
     2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct
     2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
     2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
     2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt
     2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
     2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
     2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
     2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
     2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
     2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
     2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
     3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
     3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gatctatttc cggtgaattc
     3541 gccaccatgc ccctgtccct gggagccgag atgtgggggc ctgaggcctg gctgctgctg
     3601 ctgctactgc tggcatcatt tacaggccgg tgccccgcgg gtgagctgga gacctcagac
     3661 gtggtaactg tggtgctggg ccaggacgca aaactgccct gcttctaccg aggggactcc
     3721 ggcgagcaag tggggcaagt ggcatgggct cgggtggacg cgggcgaagg cgcccaggaa
     3781 ctagcgctac tgcactccaa atacgggctt catgtgagcc cggcttacga gggccgcgtg
     3841 gagcagccgc cgcccccacg caaccccctg gacggctcag tgctcctgcg caacgcagtg
     3901 caggcggatg agggcgagta cgagtgccgg gtcagcacct tccccgccgg cagcttccag
     3961 gcgcggctgc ggctccgagt gctggtgcct cccctgccct cactgaatcc tggtccagca
     4021 ctagaagagg gccagggcct gaccctggca gcctcctgca cagctgaggg cagcccagcc
     4081 cccagcgtga cctgggacac ggaggtcaaa ggcacaacgt ccagccgttc cttcaagcac
     4141 tcccgctctg ctgccgtcac ctcagagttc cacttggtgc ctagccgcag catgaatggg
     4201 cagccactga cttgtgtggt gtcccatcct ggcctgctcc aggaccaaag gatcacccac
     4261 atcctccacg tgtccttcct tgctgaggcc tctgtgaggg gccttgaaga ccaaaatctg
     4321 tggcacattg gcagagaagg agctatgctc aagtgcctga gtgaagggca gccccctccc
     4381 tcatacaact ggacacggct ggatgggcct ctgcccagtg gggtacgagt ggatggggac
     4441 actttgggct ttcccccact gaccactgag cacagcggca tctacgtctg ccatgtcagc
     4501 aatgagttct cctcaaggga ttctcaggtc actgtggatg ttcttgaccc ccaggaagac
     4561 tctgggaagc aggtggacct agtgtcagcc tcggtggtgg tggtgggtgt gatcgccgca
     4621 ctcttgttct gccttctggt ggtggtggtg gtgctcatgt cccgatacca tcggcgcaag
     4681 gcccagcaga tgacccagaa atatgaggag gagctgaccc tgaccaggga gaactccatc
     4741 cggaggctgc attcccatca cacggacccc aggagccagc cggaggagag tgtagggctg
     4801 agagccgagg gccaccctga tagtctcaag gacaacagta gctgctctgt gatgagtgaa
     4861 gagcccgagg gccgcagtta ctccacgctg accacggtga gggagataga aacacagact
     4921 gaactgctgt ctccaggctc tgggcgggcc gaggaggagg aagatcagga tgaaggcatc
     4981 aaacaggcca tgaaccattt tgttcaggag aatgggaccc tacgggccaa gcccacgggc
     5041 aatggcatct acatcaatgg gcggggacac ctggtctgaa ctagttctag agcggccgcg
     5101 gatcccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     5161 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     5221 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     5281 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     5341 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     5401 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     5461 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     5521 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     5581 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     5641 accacgggga cgtggttttc ctttgaaaaa cacgatgata agcttgccac aacccacaag
     5701 gagacgacct tccatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     5761 cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     5821 cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     5881 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     5941 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     6001 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     6061 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     6121 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     6181 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     6241 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     6301 gcccggtgcc tagacgcgtc tggaacaatc aacctctgga ttacaaaatt tgtgaaagat
     6361 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     6421 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     6481 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     6541 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     6601 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     6661 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     6721 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     6781 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     6841 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     6901 gggccgcctc cccgcctgga attaattctg cagtcgagac ctagaaaaac atggagcaat
     6961 cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga
     7021 ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc
     7081 agctgtagat cttagccact ttttaaaaga aaagagggga ctggaagggc taattcactc
     7141 ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga
     7201 ttagcagaac tacacaccag ggccaggggt cagatatcca ctgacctttg gatggtgcta
     7261 caagctagta ccagttgagc cagataaggt agaagaggcc aataaaggag agaacaccag
     7321 cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag tgttagagtg
     7381 gaggtttgac agccgcctag catttcatca cgtggcccga gagctgcatc cggagtactt
     7441 caagaactgc tgatatcgag cttgctacaa gggactttcc gctggggact ttccagggag
     7501 gcgtggcctg ggcgggactg gggagtggcg agccctcaga tcctgcatat aagcagctgc
     7561 tttttgcctg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     7621 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     7681 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     7741 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa
     7801 agaaatgaat atcagagagt gagaggcctt gacattgcta gcgtttaccg tcgacctcta
     7861 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     7921 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     7981 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     8041 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     8101 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     8161 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     8221 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     8281 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     8341 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     8401 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     8461 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     8521 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     8581 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     8641 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     8701 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     8761 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     8821 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     8881 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     8941 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     9001 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     9061 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     9121 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     9181 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     9241 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     9301 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     9361 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     9421 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     9481 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     9541 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     9601 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     9661 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     9721 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     9781 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     9841 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     9901 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     9961 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
    10021 gaaaagtgcc acctgacgtc gacggatcgg gagatcaact tgtttattgc agcttataat
    10081 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
    10141 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat caactggata
    10201 actcaagcta accaaaatca tcccaaactt cccaccccat accctattac cactgccaat
    10261 tacctgtggt ttcatttact ctaaacctgt gattcctctg aattattttc attttaaaga
    10321 aattgtattt gttaaatatg tactacaaac ttagtagt
//
