LOCUS       V041010                 9817 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041010
VERSION     V041010
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..4671
                     /label="NCBP1(NM_002486)"
                     /note="NCBP1(NM_002486)"
                     /gene="NCBP1"
     CDS             4678..4743
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4787..4998
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5011..5279
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5306..5905
                     /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    5912..6500
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6574..6807
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6879..7000
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7019..7154
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7192..7208)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7216..7232
                     /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(7240..7270)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7285..7306
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7594..8182)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8353..9213)
                     /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(9214..9318)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9792..9808
                     /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 catgtcgcgg cggcggcaca gcgacgagaa cgacggtggg
     2341 cagcctcaca aaaggagaaa gacctctgat gcaaatgaaa ctgaagatca tttggaatct
     2401 ttaatatgta aagtaggaga aaagagtgcc tgctctttgg agagcaacct agaaggcttg
     2461 gctggtgttt tggaagctga tcttcctaac tacaagagca agatcttaag gcttctttgt
     2521 acagttgcac gcctattacc tgagaagctg acaatttata caacattagt tggactactg
     2581 aatgccagga attacaattt tggtggagaa tttgtagaag ccatgattcg tcaacttaaa
     2641 gaatcattga aagcaaacaa ttataatgaa gccgtgtatt tggtccgttt tttatctgat
     2701 cttgtgaatt gtcatgtgat tgccgcccca tcaatggttg ctatgtttga aaattttgta
     2761 agcgtaactc aggaagaaga tgtacctcag gtgcgacgag attggtatgt gtatgcattt
     2821 ctgtcatctt tgccctgggt tggaaaggag ttgtacgaaa agaaagatgc agagatggac
     2881 cgcatctttg ccaacactga aagctatctt aaaagacgcc aaaagactca tgtacccatg
     2941 ttacaggtat ggactgctga taaaccacat ccacaagaag agtatttaga ttgcctgtgg
     3001 gcccagattc agaaattgaa aaaggatcgc tggcaggaac ggcacatcct aagaccttat
     3061 cttgcctttg acagcatcct gtgtgaagca ctgcagcaca atctgcctcc ttttacacca
     3121 cctcctcaca ctgaagattc agtgtaccca atgccaaggg tcatcttcag aatgtttgat
     3181 tacacagatg atcccgaggg tcctgtcatg ccagggagtc attcagtgga aagatttgta
     3241 atagaagaga atcttcactg catcattaag tcccactgga aggaaaggaa gacttgtgct
     3301 gcacagttag tgagctatcc agggaagaac aagatcccct tgaactacca catagttgag
     3361 gtgatctttg cagagctgtt tcaacttcca gcaccccctc acattgatgt gatgtacaca
     3421 acactcctca ttgaactgtg caaacttcaa cctggctctc taccccaagt tcttgcacag
     3481 gcaactgaaa tgctatacat gcgtttggac acaatgaaca ctacctgtgt agacaggttt
     3541 attaattggt tttctcatca tctaagtaac ttccagttcc gttggagctg ggaagattgg
     3601 tcagattgtc ttagtcaaga tcctgaaagt cccaaaccga agtttgtaag agaagttcta
     3661 gaaaaatgta tgaggttgtc ttaccatcag cgtatattag atattgttcc tcctaccttc
     3721 tcagctctgt gtcctgcaaa cccaacctgc atttacaagt atggagatga aagtagcaat
     3781 tctcttcctg gacattctgt tgccctctgt ttagctgttg cctttaaaag taaggcaacc
     3841 aatgatgaaa tcttcagcat tctgaaagat gtaccaaatc ctaaccagga tgatgacgac
     3901 gatgaaggat tcagttttaa cccattgaaa atagaagtct ttgtacagac tctgctacac
     3961 ttggcagcca aatcattcag ccactccttc agtgctcttg caaagtttca tgaagtcttc
     4021 aaaaccctag ctgaaagtga tgaaggaaag ttacatgtgc taagagttat gtttgaggtc
     4081 tggaggaacc atccacagat gattgctgta ctagtggata agatgattcg tacacaaata
     4141 gttgattgtg ctgccgtagc aaattggatc ttctcttcag aactatctcg tgactttacc
     4201 agattgtttg tttgggaaat tttgcactct acaattcgta agatgaacaa acatgtcctg
     4261 aagatccaga aagagctgga agaagctaaa gagaaacttg ctaggcaaca caaacggcga
     4321 agtgatgatg acgacagaag cagtgacagg aaagacgggg ttcttgagga acaaatagaa
     4381 cgacttcagg aaaaagtgga atctgctcag agtgaacaaa agaatctttt cctcgttata
     4441 tttcagcggt ttatcatgat cttgaccgag cacctagtac gatgcgaaac tgatgggacc
     4501 agtgtattaa caccatggta taagaactgt atagagaggc tgcagcagat cttcctacag
     4561 catcaccaaa taatccagca gtacatggtg accctggaga accttctctt cactgctgaa
     4621 ttagaccctc atatcttggc cgtgttccag cagttctgtg ccctgcaggc cggatccgac
     4681 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac
     4741 aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca
     4801 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga
     4861 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     4921 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     4981 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc
     5041 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt
     5101 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc
     5161 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg
     5221 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc
     5281 aagctgtgac cggcgcctac gctagatgac cgagtacaag cccacggtgc gcctcgccac
     5341 ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     5401 cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     5461 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     5521 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     5581 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     5641 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     5701 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     5761 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     5821 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     5881 catgacccgc aagcccggtg cctgagtcga caatcaacct ctggattaca aaatttgtga
     5941 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     6001 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     6061 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     6121 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     6181 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     6241 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     6301 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     6361 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     6421 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     6481 cctttgggcc gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag
     6541 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     6601 aataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     6661 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     6721 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6781 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     6841 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     6901 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6961 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     7021 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     7081 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     7141 cttttttgga ggcctagact tttgcagaga cggcccaaat tcgtaatcat ggtcatagct
     7201 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     7261 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     7321 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     7381 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     7441 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     7501 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     7561 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7621 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7681 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7741 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7801 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7861 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7921 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7981 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     8041 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     8101 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     8161 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     8221 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     8281 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     8341 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     8401 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     8461 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     8521 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8581 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8641 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8701 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8761 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8821 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8881 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8941 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     9001 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     9061 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     9121 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     9181 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     9241 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     9301 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     9361 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     9421 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     9481 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     9541 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     9601 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     9661 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     9721 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     9781 gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
