LOCUS       V047268                 9577 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047268
VERSION     V047268
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"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..4431
                     /label="NCL(NM_005381)"
                     /note="NCL(NM_005381)"
                     /gene="NCL"
     CDS             4438..4503
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4547..4758
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4771..5039
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5066..5665
                     /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    5672..6260
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6334..6567
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6639..6760
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6779..6914
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6952..6968)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6976..6992
                     /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(7000..7030)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7045..7066
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7354..7942)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8113..8973)
                     /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(8974..9078)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9552..9568
                     /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 gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catggtgaag ctcgcgaagg caggtaaaaa tcaaggtgac
     2341 cccaagaaaa tggctcctcc tccaaaggag gtagaagaag atagtgaaga tgaggaaatg
     2401 tcagaagatg aagaagatga tagcagtgga gaagaggtcg tcatacctca gaagaaaggc
     2461 aagaaggctg ctgcaacctc agcaaagaag gtggtcgttt ccccaacaaa aaaggttgca
     2521 gttgccacac cagccaagaa agcagctgtc actccaggca aaaaggcagc agcaacacct
     2581 gccaagaaga cagttacacc agccaaagca gttaccacac ctggcaagaa gggagccaca
     2641 ccaggcaaag cattggtagc aactcctggt aagaagggtg ctgccatccc agccaagggg
     2701 gcaaagaatg gcaagaatgc caagaaggaa gacagtgatg aagaggagga tgatgacagt
     2761 gaggaggatg aggaggatga cgaggacgag gatgaggatg aagatgaaat tgaaccagca
     2821 gcgatgaaag cagcagctgc tgcccctgcc tcagaggatg aggacgatga ggatgacgaa
     2881 gatgatgagg atgacgatga cgatgaggaa gatgactctg aagaagaagc tatggagact
     2941 acaccagcca aaggaaagaa agctgcaaaa gttgttcctg tgaaagccaa gaacgtggct
     3001 gaggatgaag atgaagaaga ggatgatgag gacgaggatg acgacgacga cgaagatgat
     3061 gaagatgatg atgatgaaga tgatgaggag gaggaagaag aggaggagga agagcctgtc
     3121 aaagaagcac ctggaaaacg aaagaaggaa atggccaaac agaaagcagc tcctgaagcc
     3181 aagaaacaga aagtggaagg cacagaaccg actacggctt tcaatctctt tgttggaaac
     3241 ctaaacttta acaaatctgc tcctgaatta aaaactggta tcagcgatgt ttttgctaaa
     3301 aatgatcttg ctgttgtgga tgtcagaatt ggtatgacta ggaaatttgg ttatgtggat
     3361 tttgaatctg ctgaagacct ggagaaagcg ttggaactca ctggtttgaa agtctttggc
     3421 aatgaaatta aactagagaa accaaaagga aaagacagta agaaagagcg agatgcgaga
     3481 acacttttgg ctaaaaatct cccttacaaa gtcactcagg atgaattgaa agaagtgttt
     3541 gaagatgctg cggagatcag attagtcagc aaggatggga aaagtaaagg gattgcttat
     3601 attgaattta agacagaagc tgatgcagag aaaacctttg aagaaaagca gggaacagag
     3661 atcgatgggc gatctatttc cctgtactat actggagaga aaggtcaaaa tcaagactat
     3721 agaggtggaa agaatagcac ttggagtggt gaatcaaaaa ctctggtttt aagcaacctc
     3781 tcctacagtg caacagaaga aactcttcag gaagtatttg agaaagcaac ttttatcaaa
     3841 gtaccccaga accaaaatgg caaatctaaa gggtatgcat ttatagagtt tgcttcattc
     3901 gaagacgcta aagaagcttt aaattcctgt aataaaaggg aaattgaggg cagagcaatc
     3961 aggctggagt tgcaaggacc caggggatca cctaatgcca gaagccagcc atccaaaact
     4021 ctgtttgtca aaggcctgtc tgaggatacc actgaagaga cattaaagga gtcatttgac
     4081 ggctccgttc gggcaaggat agttactgac cgggaaactg ggtcctccaa agggtttggt
     4141 tttgtagact tcaacagtga ggaggatgcc aaagctgcca aggaggccat ggaagacggt
     4201 gaaattgatg gaaataaagt taccttggac tgggccaaac ctaagggtga aggtggcttc
     4261 gggggtcgtg gtggaggcag aggcggcttt ggaggacgag gtggtggtag aggaggccga
     4321 ggaggatttg gtggcagagg ccggggaggc tttggagggc gaggaggctt ccgaggaggc
     4381 agaggaggag gaggtgacca caagccacaa ggaaagaaga cgaagtttga aggatccgac
     4441 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac
     4501 aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca
     4561 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga
     4621 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     4681 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     4741 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc
     4801 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt
     4861 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc
     4921 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg
     4981 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc
     5041 aagctgtgac cggcgcctac gctagatgac cgagtacaag cccacggtgc gcctcgccac
     5101 ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     5161 cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     5221 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     5281 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     5341 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     5401 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     5461 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     5521 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     5581 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     5641 catgacccgc aagcccggtg cctgagtcga caatcaacct ctggattaca aaatttgtga
     5701 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     5761 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     5821 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     5881 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     5941 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     6001 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     6061 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     6121 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     6181 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     6241 cctttgggcc gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag
     6301 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     6361 aataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     6421 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     6481 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6541 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     6601 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     6661 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6721 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     6781 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     6841 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     6901 cttttttgga ggcctagact tttgcagaga cggcccaaat tcgtaatcat ggtcatagct
     6961 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     7021 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     7081 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     7141 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     7201 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     7261 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     7321 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7381 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7441 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7501 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7561 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7621 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7681 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7741 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     7801 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7861 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7921 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7981 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     8041 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     8101 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     8161 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     8221 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     8281 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8341 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8401 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8461 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8521 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8581 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8641 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8701 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8761 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8821 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8881 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8941 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     9001 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     9061 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     9121 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     9181 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     9241 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     9301 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     9361 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     9421 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     9481 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     9541 gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
