LOCUS       V046352                 8971 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046352
VERSION     V046352
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..3825
                     /label="P4HB(NM_000918)"
                     /note="P4HB(NM_000918)"
                     /gene="P4HB"
     CDS             3832..3897
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3941..4152
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4165..4433
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4460..5059
                     /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    5066..5654
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5728..5961
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6033..6154
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6173..6308
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6346..6362)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6370..6386
                     /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(6394..6424)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6439..6460
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6748..7336)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7507..8367)
                     /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(8368..8472)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8946..8962
                     /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 catgctgcgc cgcgctctgc tgtgcctggc cgtggccgcc
     2341 ctggtgcgcg ccgacgcccc cgaggaggag gaccacgtcc tggtgctgcg gaaaagcaac
     2401 ttcgcggagg cgctggcggc ccacaagtac ctgctggtgg agttctatgc cccttggtgt
     2461 ggccactgca aggctctggc ccctgagtat gccaaagccg ctgggaagct gaaggcagaa
     2521 ggttccgaga tcaggttggc caaggtggac gccacggagg agtctgacct ggcccagcag
     2581 tacggcgtgc gcggctatcc caccatcaag ttcttcagga atggagacac ggcttccccc
     2641 aaggaatata cagctggcag agaggctgat gacatcgtga actggctgaa gaagcgcacg
     2701 ggcccggctg ccaccaccct gcctgacggc gcagctgcag agtccttggt ggagtccagc
     2761 gaggtggctg tcatcggctt cttcaaggac gtggagtcgg actctgccaa gcagtttttg
     2821 caggcagcag aggccatcga tgacatacca tttgggatca cttccaacag tgacgtgttc
     2881 tccaaatacc agctcgacaa agatggggtt gtcctcttta agaagtttga tgaaggccgg
     2941 aacaactttg aaggggaggt caccaaggag aacctgctgg actttatcaa acacaaccag
     3001 ctgccccttg tcatcgagtt caccgagcag acagccccga agatttttgg aggtgaaatc
     3061 aagactcaca tcctgctgtt cttgcccaag agtgtgtctg actatgacgg caaactgagc
     3121 aacttcaaaa cagcagccga gagcttcaag ggcaagatcc tgttcatctt catcgacagc
     3181 gaccacaccg acaaccagcg catcctcgag ttctttggcc tgaagaagga agagtgcccg
     3241 gccgtgcgcc tcatcaccct ggaggaggag atgaccaagt acaagcccga atcggaggag
     3301 ctgacggcag agaggatcac agagttctgc caccgcttcc tggagggcaa aatcaagccc
     3361 cacctgatga gccaggagct gccggaggac tgggacaagc agcctgtcaa ggtgcttgtt
     3421 gggaagaact ttgaagacgt ggcttttgat gagaaaaaaa acgtctttgt ggagttctat
     3481 gccccatggt gtggtcactg caaacagttg gctcccattt gggataaact gggagagacg
     3541 tacaaggacc atgagaacat cgtcatcgcc aagatggact cgactgccaa cgaggtggag
     3601 gccgtcaaag tgcacagctt ccccacactc aagttctttc ctgccagtgc cgacaggacg
     3661 gtcattgatt acaacgggga acgcacgctg gatggtttta agaaattcct ggagagcggt
     3721 ggccaggatg gggcagggga tgatgacgat ctcgaggacc tggaagaagc agaggagcca
     3781 gacatggagg aagacgatga tcagaaagct gtgaaagatg aactgggatc cgactacaaa
     3841 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag
     3901 cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc
     3961 acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg
     4021 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
     4081 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
     4141 gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct
     4201 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc
     4261 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt
     4321 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg
     4381 cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg
     4441 tgaccggcgc ctacgctaga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     4501 cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     4561 ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     4621 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     4681 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     4741 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     4801 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     4861 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     4921 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     4981 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     5041 ccgcaagccc ggtgcctgag tcgacaatca acctctggat tacaaaattt gtgaaagatt
     5101 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     5161 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     5221 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     5281 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     5341 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     5401 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     5461 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     5521 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     5581 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     5641 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta
     5701 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag
     5761 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     5821 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     5881 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     5941 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     6001 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     6061 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6121 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     6181 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     6241 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     6301 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc
     6361 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6421 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6481 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6541 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6601 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6661 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6721 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6781 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6841 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6901 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6961 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7021 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7081 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7141 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7201 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7261 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7321 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7381 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7441 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7501 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7561 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7621 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7681 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7741 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7801 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7861 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7921 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7981 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8041 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8101 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8161 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8221 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8281 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8341 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8401 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8461 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     8521 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     8581 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     8641 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     8701 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     8761 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     8821 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     8881 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     8941 acgttgtaaa acgacggcca gtgccaagct g
//
