LOCUS       V045189                 9307 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045189
VERSION     V045189
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /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    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     enhancer        1933..2236
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2237..2440
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2475..4526
                     /label="KBTBD7(NM_032138)"
                     /note="KBTBD7(NM_032138)"
                     /gene="KBTBD7"
     CDS             4533..4598
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             4608..4664
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /transl_table=1
                     /translation="ATNFSLLKQAGDVEENPGP"
                     /label="P2A"
     CDS             4677..5396
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     misc_feature    5403..5990
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6064..6297
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6369..6490
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6509..6644
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6682..6698)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6706..6722
                     /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(6730..6760)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6775..6796
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7084..7672)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7843..8703)
                     /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(8704..8808)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9282..9298
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     1981 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     2041 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     2101 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     2161 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     2221 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     2281 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     2341 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     2401 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     2461 ccgctagcgc caccatgcag tcccgggaag acgtcccgcg ctctcgccgc ctcgccagtc
     2521 cccgtggtgg gaggcggccc aagaggattt ccaagccctc ggtttcggcc tttttcacgg
     2581 gtccagagga gttaaaggac acggcccatt ctgcagccct gctggcacag ctcaagtcct
     2641 tctacgacgc gcggctgctg tgtgatgtga ccatcgaggt ggtgacgcct ggcagcgggc
     2701 ctggcacggg tcgcctcttt tcctgcaatc gcaacgtgct agcagctgcg tgtccctact
     2761 tcaagagcat gttcacaggt ggcatgtacg agagccagca ggccagcgtg accatgcacg
     2821 atgtggacgc cgagtccttc gaggtgttgg tcgactactg ctacacgggt cgtgtgtctc
     2881 tcagtgaggc caatgtgcag cgcctgtacg cggcctccga catgctacag ctggaatatg
     2941 tgcgggaagc ctgtgcctcc ttcttagccc gacgtcttga cctgaccaac tgcaccgcca
     3001 tcctcaagtt tgcagacgcc ttcgaccatc acaagcttcg atctcaggcc cagtcctaca
     3061 tagctcacaa cttcaagcag ctcagccgaa tgggttcaat tcgggaggag actctagcag
     3121 atctaaccct ggcccagctg ctggctgtcc tacgcctgga tagtctggac atagagagtg
     3181 agcggactgt atgccatgta gctgtgcagt ggctggaggc tgctgccaaa gagcggggtc
     3241 ccagtgctgc agaagtcttc aagtgcgtgc gctggatgca cttcactgaa gaagatcagg
     3301 actacttaga agggctgctg accaagccca tcgtgaagaa gtactgcctg gacgttattg
     3361 aaggggccct gcagatgcgc tatggtgacc tgttgtacaa gtctctggtg ccagtgccaa
     3421 acagcagcag cagcagtagc agcagcaact ctcttgtatc tgcagcagaa aatccacccc
     3481 agagactggg tatgtgtgcc aaggagatgg tgatcttctt tggacatcct agagatccct
     3541 ttctctgcta tgacccttac tcgggggaca tttacacaat gccatcccct ttgaccagct
     3601 ttgctcacac taagactgtc acctcctcag ctgtctgtgt gtccccagac catgacatct
     3661 atctagctgc tcagcccagg aaagacctct gggtgtataa accagctcag aatagttggc
     3721 agcaacttgc agatcgcttg ctgtgtcgtg agggcatgga tgtggcatat ctcaatggct
     3781 acatctacat tttgggggga cgagacccta ttactggagt taagttgaag gaagtggaat
     3841 gctacagtgt tcagagaaac cagtgggcat tggtggctcc tgtccctcat tccttctatt
     3901 cctttgaact catagtggtt cagaactatc tttatgctgt caacagtaag cgcatgcttt
     3961 gctatgatcc tagccacaat atgtggctga actgtgcttc tcttaaacgt agtgactttc
     4021 aggaagcatg tgtcttcaat gatgaaatct attgtatctg tgacatccca gtcatgaagg
     4081 tctacaaccc agctagggga gaatggaggc ggattagtaa tattcctttg gattcagaga
     4141 cccacaacta ccagattgtc aatcatgacc aaaagttgct tctcatcact tctacaaccc
     4201 cacaatggaa aaagaaccga gtgacagtgt atgagtatga tactagggaa gatcagtgga
     4261 ttaatatagg taccatgtta ggccttttgc agtttgactc tggctttatt tgcctttgtg
     4321 ctcgtgttta tccttcctgc cttgaacctg gtcagagttt tattactgag gaagatgatg
     4381 cacggagtga gtctagtact gaatgggact tagatggatt cagtgagctg gactctgagt
     4441 caggaagttc aagttctttt tcagatgatg aagtctgggt gcaagtagca cctcagcgaa
     4501 atgcacagga tcagcagggt tctttggtcg acgactacaa agaccatgac ggtgattata
     4561 aagatcatga catcgactac aaggatgacg atgacaaggg aagcggagcc actaacttct
     4621 ccctgttgaa acaagcaggg gatgtcgaag agaatcccgg gccagaattc gccaccatgg
     4681 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     4741 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     4801 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     4861 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     4921 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     4981 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     5041 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     5101 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     5161 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     5221 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     5281 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     5341 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaggat
     5401 ccaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     5461 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     5521 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     5581 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     5641 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     5701 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     5761 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     5821 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     5881 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     5941 tccgccttcg ccctcagacg agtcggatct ccctttggcc gcctccccgc ctggtacctt
     6001 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     6061 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt
     6121 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     6181 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     6241 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     6301 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     6361 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6421 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6481 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca
     6541 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     6601 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga
     6661 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6721 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6781 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6841 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6901 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6961 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     7021 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     7081 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     7141 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     7201 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     7261 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7321 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7381 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7441 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7501 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     7561 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7621 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7681 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7741 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7801 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7861 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7921 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7981 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     8041 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     8101 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     8161 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     8221 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     8281 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8341 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8401 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8461 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8521 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8581 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8641 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8701 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8761 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8821 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     8881 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     8941 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     9001 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     9061 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     9121 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     9181 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     9241 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
     9301 caagctg
//
