LOCUS       V029013                 9601 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029013
VERSION     V029013
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..4254
                     /label="Pias1(NM_019663)"
                     /note="Pias1(NM_019663)"
                     /gene="Pias1"
     CDS             4261..4326
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4370..4581
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4594..4862
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4895..5689
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     misc_feature    5696..6284
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6358..6591
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6663..6784
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6803..6938
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6976..6992)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7000..7016
                     /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(7024..7054)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7069..7090
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7378..7966)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8137..8997)
                     /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(8998..9102)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9576..9592
                     /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 catggcggac agtgcggaac taaagcaaat ggttatgagc
     2341 cttagagttt ctgaactcca agtactgttg ggctacgctg ggaggaacaa gcacggacgc
     2401 aaacacgaac ttcttacaaa agccctgcat ttgttaaagg ctggctgtag tcctgctgta
     2461 caaatgaaaa ttaaagaact ctacaggagg cggttccctc agaaaattat gacgcctgcg
     2521 gacttgtcta tccccaacgt acattcaagt cctatgcctc cgactctttc tccatccacc
     2581 attccacagc tcacttatga tggccaccct gcatcatccc cactactccc tgtttctctt
     2641 ctgggaccca aacatgaact ggaactccca catctcacgt cagcgctgca cccagtccac
     2701 ccggacataa agctgcagaa gctaccattc tatgacctgt tggatgaact gatcaagccc
     2761 accagtctag cttcagacaa cagccagcgc tttcgggaaa cctgttttgc atttgccttg
     2821 acaccacaac aggtgcagca gatcagcagc tccatggata tttctgggac caaatgtgac
     2881 ttcacagtgc aggtccaatt aaggttttgt ttatcagaaa ccagttgtcc acaagaagat
     2941 cacttcccac ccaacctttg tgtaaaagtg aatacaaaac cttgcagcct tccaggttac
     3001 cttccaccta ctaaaaacgg tgtggaacca aagcgaccta gccgaccaat taatatcacc
     3061 tcacttgtcc gattgtccac gacagtacca aataccattg ttgtttcttg gactgcagaa
     3121 attggaagaa cctattccat ggcagtatat cttgtaaaac agttgtcctc aacagttctt
     3181 cttcagaggt tacgagcaaa gggaataagg aatccggatc attctagagc tttaattaaa
     3241 gagaagttaa ctgcagatcc agatagtgag atagctacta ccagcctacg ggtttcgctg
     3301 ctgtgtccac ttgggaaaat gcgactgaca atcccctgtc gggcacttac ctgctcccac
     3361 cttcagtgtt ttgatgcaac tctttacatt caaatgaatg agaaaaaacc aacatgggtt
     3421 tgtcctgtct gtgataagaa ggccccatat gaacacctta ttattgacgg gttgtttatg
     3481 gaaattctaa agtactgcac agactgtgac gagatacagt ttaaggagga tggctcgtgg
     3541 gctccaatga ggtcaaagaa ggaggttcaa gaagtcactg cctcctacaa tggagttgat
     3601 ggttgcttga gctccacatt ggagcatcag gtagcgtccc acaaccagtc ctcaaataaa
     3661 aacaagaaag tcgaggtcat tgacctaacc attgacagct cgtcagatga agaggaggaa
     3721 gaaccccctg ccaagaggac ctgtccttcc ctgtctccta cgtcaccact aagtaataaa
     3781 ggcattttaa gtcttcctca tcaagcctcg cctgtgtccc gcaccccaag ccttcctgct
     3841 gtagatacaa gctacatcaa cacctccctc atccaggact acaggcaccc cttccacatg
     3901 acgcctatgc cttatgactt acaaggatta gatttctttc ctttcttatc aggagacaat
     3961 cagcattaca acacctccct gctagccgca gctgcagcgg cggtctcaga tgaccaggac
     4021 ctcctgcact cctcccggtt tttcccgtat acctcctcgc agatgtttct cgaccagcta
     4081 agtgcaggag ggagcacatc tctgccagcc accaacggaa gcagtagcgg cagcaacagc
     4141 agccttgtgt cttccaacag tctgagagag agccatggcc atggtgtggc cagcaggagc
     4201 agcgcagaca cagcgtccat ctttggcatc ataccagaca ttatctcatt ggacggatcc
     4261 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat
     4321 gacaagtagc ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc
     4381 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag
     4441 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     4501 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
     4561 gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc
     4621 cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc
     4681 tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg
     4741 ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc
     4801 ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga
     4861 tccaagctgt gaccggcgcc tacgctaggc caccatggct gaacaagatg gattgcacgc
     4921 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4981 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     5041 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     5101 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5161 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5221 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5281 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5341 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5401 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5461 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5521 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5581 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5641 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag tcgacaatca
     5701 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     5761 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     5821 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     5881 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     5941 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     6001 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     6061 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     6121 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     6181 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     6241 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac
     6301 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     6361 aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact gggtctctct
     6421 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     6481 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     6541 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     6601 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     6661 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6721 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6781 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg cccattctcc
     6841 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     6901 gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca gagacggccc
     6961 aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7021 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     7081 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7141 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     7201 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7261 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7321 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7381 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7441 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7501 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7561 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7621 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7681 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7741 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7801 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7861 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7921 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7981 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     8041 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     8101 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     8161 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     8221 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     8281 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     8341 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8401 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     8461 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     8521 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     8581 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8641 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8701 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     8761 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8821 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8881 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8941 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     9001 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     9061 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     9121 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     9181 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     9241 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     9301 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     9361 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     9421 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     9481 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     9541 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct
     9601 g
//
