LOCUS       V023051                10243 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023051
VERSION     V023051
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"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2888..4849
                     /label="CNTN6(NM_001349359)"
                     /note="CNTN6(NM_001349359)"
                     /gene="CNTN6"
     CDS             4850..4915
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4946..5532
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             5552..6151
                     /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    6165..6753
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6960..7593
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7721..7737)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7745..7761
                     /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(7769..7799)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7814..7835
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8123..8711)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8882..9742)
                     /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(9743..9847)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9895..10029
                     /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 cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tctatttccg gtgaattcgt atacatggtt
     2821 aaaaaatggt gaacgactca acccagagga gagaattcaa atagaaaatg ggacactcat
     2881 cataacgatg ctgaatgtgt cagattctgg tgtgtaccaa tgtgctgcag aaaacaaata
     2941 tcagataatt tatgcaaatg ctgaattgag agttttagcc tcagctccag atttctccaa
     3001 aagtccagtt aaaaaaaagt cttttgttca agttggtggg gatattgtta tcggatgcaa
     3061 accaaatgct tttcccaggg cagctatctc ttggaaaaga ggaacggaga cccttagaca
     3121 aagcaaaaga atatttctct tggaggatgg cagcctcaag atatataata ttaccaggtc
     3181 agatgctgga tcatatacat gcatagccac aaatcagttt ggcactgcaa agaacactgg
     3241 cagcctcatt gtaaaagaga gaactgtcat taccgtccca ccttccaaaa tggatgttac
     3301 agttggcgag agtatagtgc taccatgcca ggtgtcccat gacccctcca ttgaagtggt
     3361 atttgtatgg tttttcaatg gagatgtcat agacttaaaa aaaggagtgg ctcattttga
     3421 aaggattgga ggagaatctg ttggggattt gatgataagg aatattcagt tacatcattc
     3481 aggaaaatat ctctgcacag tacaaacaac cctagaaagt ttatctgcag tagccgatat
     3541 cattgttaga ggtccaccag gtcctcctga ggatgtgcaa gtggaagaca tttccagtac
     3601 tacttctcaa ctaagttgga gagcaggccc agataataac agtcccattc aaatatttac
     3661 tattcagact cggacaccat tttctgtggg ttggcaggct gttgctacag ttccagaaat
     3721 tctcaatggt aagacataca atgcaacagt ggttggtttg agtccttggg tggaatatga
     3781 atttcgtgtt gttgccggca acagcattgg gattggagaa ccaagtgaac catcagaatt
     3841 gttaagaact aaagcatcag tccctgttgt ggcaccagta aacatccatg gaggtggagg
     3901 aagtcggtct gaactcgtca ttacgtggga gtcaattcca gaagaactgc agaatgggga
     3961 gggatttgga tatatcatca tgttccggcc agtgggctcg acaacctggt ccaaggagaa
     4021 agtgtcatct gtggaatcat caaggtttgt ctacagaaat gaaagcatca tcccactgtc
     4081 tccctttgaa gtcaaagtgg gtgtgtataa taatgaagga gaaggatccc tgagtactgt
     4141 gaccattgtc tactctgggg aagatgaacc tcaactggcc ccaaggggaa cttctctcca
     4201 gagtttttct gcttctgaaa tggaggtttc atggaatgct attgcctgga atagaaacac
     4261 tggaagagtg ctgggctatg aggtcttata ctggacagat gactccaaag aatccatgat
     4321 aggtaaaatt agagtcagtg gaaatgtcac aaccaaaaac atcacggggc tgaaagctaa
     4381 taccatctac tttgcttccg taagagctta caacactgct gggacagggc cctcaagccc
     4441 cccagtcaat gttaccacca aaaagtctcc tccaagccaa ccaccagcaa acattgcctg
     4501 gaagctgaca aactctaaat tatgcttgaa ctgggagcat gtaaaaacca tggaaaatga
     4561 gtctgaagtt ttggggtaca agattctgta ccggcaaaac agacagagta aaactcatat
     4621 tttggaaaca aacaatacat cagctgagct tctggttcca tttgaagaag actacttaat
     4681 tgaaataaga acagtcagtg atggtggaga tggaagcagc agtgaggaaa ttaggattcc
     4741 aaaaatgtca agtttgagtt ccagaggaat tcaattctta gaacctagca cccattttct
     4801 ttccattgtc attgtgattt ttcactgttt tgctattcag ccacttatcg actacaaaga
     4861 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtgact
     4921 agttctagag cggccgcgga tcccgcccct ctccctcccc cccccctaac gttactggcc
     4981 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     5041 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     5101 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
     5161 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     5221 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     5281 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     5341 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     5401 tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa
     5461 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataag
     5521 cttgccacaa cccacaagga gacgaccttc catgaccgag tacaagccca cggtgcgcct
     5581 cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta
     5641 ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca
     5701 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     5761 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     5821 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     5881 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     5941 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     6001 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     6061 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     6121 ctggtgcatg acccgcaagc ccggtgccta gacgcgtctg gaacaatcaa cctctggatt
     6181 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     6241 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     6301 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     6361 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     6421 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     6481 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     6541 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
     6601 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     6661 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     6721 cgagtcggat ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct
     6781 agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg
     6841 gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag
     6901 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact
     6961 ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac
     7021 acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact
     7081 gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa
     7141 taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga
     7201 gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga
     7261 gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc
     7321 tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc
     7381 ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag
     7441 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     7501 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     7561 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     7621 agtatttata acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc
     7681 gtttaccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     7741 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     7801 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     7861 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     7921 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     7981 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     8041 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     8101 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     8161 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     8221 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     8281 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     8341 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     8401 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     8461 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     8521 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     8581 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     8641 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     8701 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     8761 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     8821 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     8881 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     8941 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     9001 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     9061 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     9121 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     9181 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     9241 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     9301 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     9361 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     9421 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     9481 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     9541 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     9601 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     9661 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     9721 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     9781 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     9841 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatcaacttg
     9901 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     9961 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
    10021 gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc caccccatac
    10081 cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa
    10141 ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagttttt
    10201 aaagaaattg tatttgttaa atatgtacta caaacttagt agt
//
