LOCUS       V045195                 8857 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045195
VERSION     V045195
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..4076
                     /label="KBTBD4(NM_018095)"
                     /note="KBTBD4(NM_018095)"
                     /gene="KBTBD4"
     CDS             4083..4148
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             4158..4214
                     /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             4227..4946
                     /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    4953..5540
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5614..5847
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5919..6040
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6059..6194
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6232..6248)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6256..6272
                     /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(6280..6310)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6325..6346
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6634..7222)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7393..8253)
                     /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(8254..8358)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8832..8848
                     /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 caccatgaaa ggagggaacg cagacagctg gcagagagag aagttggcta
     2521 gcatggaatc accagaggag cctggagcat ccatggatga gaactacttt gtgaactaca
     2581 ctttcaaaga tcggtcacat tcaggccgtg tggctcaagg catcatgaaa ctgtgtctag
     2641 aggaggagct ctttgctgat gtcaccattt cggtggaagg ccgggagttt cagctccatc
     2701 ggctggtcct ctcagctcag agctgcttct tccgatccat gttcacttcc aacctgaagg
     2761 aggcccacaa ccgggtgatt gtgctgcagg atgtcagcga gtctgttttc cagctcctgg
     2821 ttgattatat ctaccatggg actgtgaaac ttcgagctga ggagttgcag gaaatttatg
     2881 aggtgtcaga catgtatcag ctgacatctc tctttgagga atgctctcgg tttttggccc
     2941 gcacagtgca agtgggaaac tgccttcagg tgatgtggct ggcagatcgg cacagtgatc
     3001 ctgagctcta tacggctgcc aagcactgtg ccaagaccca cctggcccag ctgcagaata
     3061 cagaggaatt tctccacttg ccccaccgct tactcacaga tatcatctcg gatggagttc
     3121 cgtgttctca gaacccaaca gaggcaatag aagcctggat caactttaat aaagaggaaa
     3181 gagaggcttt tgcagagtca ctcaggacaa gcttgaagga aattggggag aatgtgcaca
     3241 tttacctgat tgggaaagag tcatctcgta cccactcgtt ggctgtgtcc ttgcactgtg
     3301 cagaagatga ctccatcagt gtaagtggcc aaaacagttt gtgccaccag atcactgcgg
     3361 cctgcaagca tggtggagac ttgtatgtgg tgggagggtc catcccacgg cgcatgtgga
     3421 agtgcaacaa tgccaccgtt gactgggagt ggtgtgctcc tttgcctcgg gaccggctcc
     3481 agcacaccct ggtgtctgtg cccgggaaag atgccatata ttcactgggt ggcaagacac
     3541 tgcaagatac cctctccaac gcagtcattt attatcgcgt aggtgataat gtgtggacag
     3601 agacaactca gctagaggtg gctgtgtcag gggctgctgg tgccaacctc aacgggatca
     3661 tctacttact agggggggag gagaatgatc tggacttctt taccaaacct tcccgactca
     3721 tccagtgctt tgacacagag acagacaaat gccatgtgaa gccctatgtg ctgccctttg
     3781 caggccgcat gcacgcagct gtgcataaag atctggtgtt catcgtggct gaaggggact
     3841 ccctggtgtg ctacaatccc ttgctagaca gcttcacccg gctttgcctt cctgaggcct
     3901 ggagctctgc cccatccctc tggaagattg ccagctgtaa cgggagcatc tatgtcttcc
     3961 gggaccgata taaaaagggg gatgccaaca cctacaagct tgaccctgcc acttcagccg
     4021 taactgtcac aagaggtatt aaggtgctgc ttaccaattt gcagtttgtg ttggccgtcg
     4081 acgactacaa agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg
     4141 atgacaaggg aagcggagcc actaacttct ccctgttgaa acaagcaggg gatgtcgaag
     4201 agaatcccgg gccagaattc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     4261 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     4321 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     4381 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     4441 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     4501 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     4561 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     4621 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     4681 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     4741 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     4801 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     4861 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     4921 tcggcatgga cgagctgtac aagtaaggat ccaatcaacc tctggattac aaaatttgtg
     4981 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     5041 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     5101 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     5161 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     5221 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     5281 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     5341 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     5401 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     5461 tgctgccggc tctgcggcct cttccgcgtc tccgccttcg ccctcagacg agtcggatct
     5521 ccctttggcc gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag
     5581 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     5641 aataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     5701 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     5761 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     5821 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     5881 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     5941 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6001 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     6061 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     6121 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     6181 cttttttgga ggcctagact tttgcagaga cggcccaaat tcgtaatcat ggtcatagct
     6241 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6301 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6361 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6421 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6481 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6541 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6601 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6661 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6721 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6781 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6841 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6901 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6961 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7021 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     7081 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7141 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7201 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7261 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7321 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7381 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7441 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7501 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7561 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7621 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7681 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7741 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7801 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7861 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7921 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7981 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8041 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8101 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8161 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8221 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8281 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8341 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     8401 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     8461 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     8521 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     8581 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     8641 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     8701 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     8761 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     8821 gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
