LOCUS       V021280                 8847 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021280
VERSION     V021280
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             2815..3498
                     /label="RNF114(NM_018683)"
                     /note="RNF114(NM_018683)"
                     /gene="RNF114"
     misc_feature    3518..4090
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES"
     CDS             4092..4802
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK*"
                     /label="mCherry"
     misc_feature    4816..5404
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5611..6244
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6372..6388)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6396..6412
                     /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(6420..6450)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6465..6486
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6774..7362)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7533..8393)
                     /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(8394..8498)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8546..8680
                     /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 gtgaattcgc caccatggcg
     2821 gcgcaacagc gggactgcgg gggtgctgcg cagctggcgg ggccggcggc ggaggctgac
     2881 cccctaggac gcttcacgtg tcccgtgtgc ttagaggtgt acgagaagcc ggtacaggtg
     2941 ccctgcggac acgtcttttg ctctgcatgc ctgcaggaat gtctgaagcc gaagaagcct
     3001 gtctgtgggg tgtgtcgcag cgctctggca cctggcgtcc gagccgtgga gctcgagcgg
     3061 cagatcgaga gcacagagac ttcttgccat ggctgccgta agaatttctt cctgtccaag
     3121 atccggtccc acgtggctac ttgttccaaa taccagaatt acatcatgga aggtgtgaag
     3181 gccaccatta aggatgcatc tcttcagcca aggaatgttc caaaccgtta cacctttcct
     3241 tgtccttact gtcctgagaa gaactttgat caggaaggac ttgtggaaca ctgcaaatta
     3301 ttccatagca cggataccaa atctgtggtt tgtccgatat gtgcctcgat gccctgggga
     3361 gaccccaact accgcagcgc caacttcaga gagcacatcc agcgccggca ccggttttct
     3421 tatgacactt ttgtggatta tgatgttgat gaagaggaca tgatgaatca ggtgttgcag
     3481 cgctccatca tcgaccagtg agcggccgcg gatcccgccc ctctccctcc ccccccccta
     3541 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     3601 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     3661 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     3721 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     3781 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     3841 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     3901 aaagagtcaa atggctcacc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     3961 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     4021 cgaggttaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     4081 acgatgataa tatggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
     4141 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
     4201 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
     4261 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
     4321 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
     4381 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
     4441 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     4501 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     4561 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     4621 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     4681 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     4741 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt
     4801 gaacgcgtct ggaacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     4861 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     4921 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     4981 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     5041 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     5101 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     5161 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     5221 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     5281 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     5341 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     5401 ccgcctggaa ttaattctgc agtcgagacc tagaaaaaca tggagcaatc acaagtagca
     5461 atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg
     5521 gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     5581 ttagccactt tttaaaagaa aagaggggac tggaagggct aattcactcc caacgaagac
     5641 aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact
     5701 acacaccagg gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac
     5761 cagttgagcc agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc
     5821 ctgtgagcct gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca
     5881 gccgcctagc atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct
     5941 gatatcgagc ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg
     6001 gcgggactgg ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt
     6061 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     6121 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     6181 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     6241 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     6301 tcagagagtg agaggccttg acattgctag cgtttaccgt cgacctctag ctagagcttg
     6361 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     6421 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     6481 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     6541 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     6601 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     6661 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6721 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6781 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6841 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6901 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6961 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7021 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7081 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7141 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7201 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7261 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7321 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7381 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7441 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7501 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7561 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7621 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7681 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7741 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7801 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7861 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7921 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7981 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     8041 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8101 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     8161 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8221 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8281 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     8341 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8401 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8461 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8521 cctgacgtcg acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata
     8581 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     8641 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa
     8701 ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt
     8761 tcatttactc taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg
     8821 ttaaatatgt actacaaact tagtagt
//
