LOCUS       V021420                 9471 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021420
VERSION     V021420
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             2301..4325
                     /label="NEXN(NM_144573)"
                     /note="NEXN(NM_144573)"
                     /gene="NEXN"
     CDS             4332..4397
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4441..4652
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4665..4933
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4960..5559
                     /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    5566..6154
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6228..6461
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6533..6654
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6673..6808
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6846..6862)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6870..6886
                     /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(6894..6924)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6939..6960
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7248..7836)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8007..8867)
                     /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(8868..8972)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9446..9462
                     /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 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta
     2281 gagctagcga attcgccacc atgaatgata tttcccaaaa ggctgagatt ctgctttctt
     2341 catctaaacc tgtcccaaaa acctatgtac caaaacttgg caagggtgat gtaaaggata
     2401 agtttgaagc catgcagaga gccagggaag aaagaaatca aaggagatct agagacgaaa
     2461 aacaaagaag aaaagaacaa tatattagag agagagaatg gaacaggaga aagcaggaga
     2521 ttaaagaaat gcttgcttct gatgatgagg aagatgtatc ttctaaagta gaaaaggctt
     2581 atgttccaaa attaacagga actgtgaagg gtagatttgc tgaaatggag aaacaaagac
     2641 aagaggaaca aaggaagaga acggaggagg aacgaaaacg cagaattgag caggatatgt
     2701 tagaaaagag gaaaatacag cgtgaattag caaaaagggc tgaacagatt gaggacataa
     2761 acaatacggg aactgaatca gcatcagagg aaggagatga ttcactactt ataactgtgg
     2821 tacctgtcaa atcatataaa acatctggaa aaatgaaaaa gaattttgag gatctagaaa
     2881 aagaacgtga agagaaagaa aggatcaagt acgaggaaga taaaagaata agatatgaag
     2941 aacaacgacc atctctcaag gaagcaaagt gtctttcatt agttatggat gatgaaatag
     3001 aaagtgaagc aaaaaaagaa tcactttctc ccggaaaatt gaaactaact tttgaagaac
     3061 tggagcgaca aagacaagaa aaccgaaaga agcaagctga agaggaagca agaaaacgtt
     3121 tagaagaaga gaagcgtgct tttgaagaag caaggcggca aatggtaaat gaagatgagg
     3181 aaaaccaaga cacagcaaaa atttttaaag ggtaccgccc tggtaaactc aaactcagtt
     3241 ttgaagaaat ggaaaggcaa agaagagaag atgaaaaaag gaaagcagaa gaagaagcca
     3301 gaaggagaat agaggaagaa aagaaggcgt ttgctgaagc aaggagaaat atggtagtag
     3361 atgatgactc cccagagatg tataagacaa tctctcaaga atttcttaca ccgggaaaac
     3421 tggaaattaa ttttgaagaa ttattaaaac aaaaaatgga agaagaaaaa cgacgaacag
     3481 aggaggaacg gaagcataag ctagaaatgg agaaacaaga atttgaacaa ctgagacagg
     3541 aaatgggaga ggaagaggaa gaaaatgaaa cctttggatt gagcagagaa tatgaagaac
     3601 tgatcaaatt aaaaaggagt ggctctattc aagctaaaaa cctaaaaagc aagtttgaaa
     3661 aaattggaca gttgtctgaa aaagaaatac agaaaaaaat agaagaagag cgagcaagaa
     3721 ggagagcaat tgaccttgaa attaaagagc gagaagctga aaattttcat gaggaagatg
     3781 atgttgatgt taggcctgca agaaaaagcg aggctccatt tactcacaaa gtgaatatga
     3841 aagctagatt tgaacaaatg gctaaggcaa gagaagaaga agaacaaaga agaattgaag
     3901 aacaaaagtt actacgcatg cagtttgaac aaagggaaat tgatgcagca ctacaaaaga
     3961 aaagagaaga ggaggaggag gaagaaggta gcatcatgaa tggctccact gctgaagatg
     4021 aagagcaaac cagatcagga gctccatggt tcaagaagcc tcttaaaaac acatcagttg
     4081 tagacagtga gccagtcaga tttacggtta aagtaacagg agaacccaaa ccagaaatta
     4141 catggtggtt tgaaggagaa atactgcagg atggagaaga ctatcaatat attgaaaggg
     4201 gagaaactta ctgcctttac ttaccagaaa ctttcccaga agatggagga gagtatatgt
     4261 gtaaagcagt caacaataaa ggatctgcag ctagtacctg tattcttacc attgaaagta
     4321 agaatggatc cgactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca
     4381 aggatgacga tgacaagtag cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt
     4441 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     4501 cgggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     4561 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     4621 tttttcgcaa cgggtttgcc gccagaacac agctgaagct tcgaggggct cgcatctctc
     4681 cttcacgcgc ccgccgccct acctgaggcc gccatccacg ccggttgagt cgcgttctgc
     4741 cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca
     4801 ggtcgagacc gggcctttgt ccggcgctcc cttggagcct acctagactc agccggctct
     4861 ccacgctttg cctgaccctg cttgctcaac tctacgtctt tgtttcgttt tctgttctgc
     4921 gccgttacag atccaagctg tgaccggcgc ctacgctaga tgaccgagta caagcccacg
     4981 gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc
     5041 gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga gcgggtcacc
     5101 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     5161 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     5221 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     5281 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     5341 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     5401 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     5461 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     5521 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag tcgacaatca acctctggat
     5581 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     5641 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     5701 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     5761 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     5821 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     5881 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     5941 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     6001 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     6061 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     6121 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta
     6181 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     6241 tcactcccaa cgaaaataag atctgctttt tgcttgtact gggtctctct ggttagacca
     6301 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     6361 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     6421 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
     6481 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt
     6541 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6601 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6661 ctggctctag ctatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     6721 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     6781 aagtagtgag gaggcttttt tggaggccta gacttttgca gagacggccc aaattcgtaa
     6841 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6901 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6961 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     7021 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     7081 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     7141 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     7201 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     7261 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     7321 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     7381 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     7441 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     7501 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7561 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7621 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7681 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7741 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7801 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7861 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7921 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7981 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     8041 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     8101 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     8161 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     8221 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     8281 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     8341 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     8401 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     8461 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     8521 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     8581 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     8641 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8701 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8761 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8821 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8881 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8941 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     9001 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     9061 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     9121 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     9181 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     9241 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     9301 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     9361 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     9421 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct g
//
