LOCUS       V027786                 9261 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027786
VERSION     V027786
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..4115
                     /label="USP2(NM_004205)"
                     /note="USP2(NM_004205)"
                     /gene="USP2"
     CDS             4122..4187
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4231..4442
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4455..4723
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4750..5349
                     /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    5356..5944
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6018..6251
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6323..6444
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6463..6598
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6636..6652)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6660..6676
                     /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(6684..6714)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6729..6750
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7038..7626)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7797..8657)
                     /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(8658..8762)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9236..9252
                     /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 atgtcccagc tctcctccac cctgaagcgc tacacagaat
     2341 cggcccgcta cacagatgcc cactatgcca agtcgggcta tggtgcctac accccgtcct
     2401 cctatggggc caatctggct gcctccttac tggagaagga gaaacttggt ttcaagccgg
     2461 tccccaccag cagcttcctc acccgtcccc gtacctatgg cccctcctcc ctcctggact
     2521 atgaccgggg ccgccccctg ctgagacccg acatcactgg gggtggtaag cgggcagaga
     2581 gccagacccg gggtactgag cggcctttag gcagtggcct cagcgggggc agcggattcc
     2641 cttatggagt gaccaacaac tgcctcagct acctgcccat caatgcctat gaccaggggg
     2701 tgaccctaac ccagaagctg gacagccaat cagacctggc ccgggatttc tccagcctcc
     2761 ggacctcaga tagctaccgg atagacccca ggaacctggg ccgcagcccc atgctggccc
     2821 ggacgcgcaa ggagctctgc accctgcagg ggctctacca gacagccagc tgccctgaat
     2881 acctggtcga ctacctggag aactatggtc gcaagggcag tgcatctcag gtgccctccc
     2941 aggcccctcc ctcacgagtc cctgaaatca tcagcccaac ctaccgaccc attggccgct
     3001 acacgctgtg ggagacggga aagggtcagg cccctgggcc cagccgctcc agctccccgg
     3061 gaagagacgg catgaattct aagagtgccc agggtctggc tggtcttcga aaccttggga
     3121 acacgtgctt catgaactca attctgcagt gcctgagcaa cactcgggag ttgagagatt
     3181 actgcctcca gaggctctac atgcgggacc tgcaccacgg cagcaatgca cacacagccc
     3241 tcgtggaaga gtttgcaaaa ctaattcaga ccatatggac ttcatccccc aatgatgtgg
     3301 tgagcccatc tgagttcaag acccagatcc agagatacgc accgcgcttt gttggctata
     3361 atcagcagga tgctcaggag ttccttcgct ttcttctgga tgggctccat aacgaggtga
     3421 accgagtgac actgagacct aagtccaacc ctgagaacct cgatcatctt cctgatgacg
     3481 agaaaggccg acagatgtgg agaaaatatc tagaacggga agacagtagg atcggggatc
     3541 tctttgttgg gcagctaaag agctcgctga cgtgtacaga ttgtggttac tgttctacgg
     3601 tcttcgaccc cttctgggac ctctcactgc ccattgctaa gcgaggttat cctgaggtga
     3661 cattaatgga ctgcatgagg ctcttcacca aagaggatgt gcttgatgga gatgaaaagc
     3721 caacatgctg tcgctgccga ggcagaaaac ggtgtataaa gaagttctcc atccagaggt
     3781 tcccaaagat cttggtgctc catctgaagc ggttctcaga atccaggatc cgaaccagca
     3841 agctcacaac atttgtgaac ttccccctaa gagacctgga cttaagagaa tttgcctcag
     3901 aaaacaccaa ccatgctgtt tacaacctgt acgctgtgtc caatcactcc ggaaccacca
     3961 tgggtggcca ctatacagcc tactgtcgca gtccagggac aggagaatgg cacactttca
     4021 acgactccag cgtcactccc atgtcctcca gccaagtgcg caccagcgac gcctacctgc
     4081 tcttctacga actggccagc ccgccctccc gaatgggatc cgactacaaa gaccatgacg
     4141 gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag cggccgcgaa
     4201 ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
     4261 agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa
     4321 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
     4381 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
     4441 agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc
     4501 gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg
     4561 cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc
     4621 cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac
     4681 tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg tgaccggcgc
     4741 ctacgctaga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     4801 cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     4861 gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     4921 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     4981 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     5041 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     5101 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     5161 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     5221 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     5281 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     5341 ggtgcctgag tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     5401 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     5461 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     5521 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     5581 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     5641 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     5701 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     5761 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     5821 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     5881 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     5941 ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     6001 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag atctgctttt
     6061 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     6121 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     6181 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     6241 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
     6301 atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag
     6361 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     6421 gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg
     6481 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     6541 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     6601 gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6661 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6721 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6781 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6841 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6901 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6961 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     7021 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     7081 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7141 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7201 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     7261 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7321 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7381 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7441 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     7501 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7561 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7621 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7681 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7741 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7801 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7861 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7921 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7981 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     8041 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     8101 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     8161 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     8221 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     8281 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     8341 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     8401 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     8461 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8521 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     8581 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8641 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8701 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8761 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     8821 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     8881 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     8941 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     9001 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     9061 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
     9121 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     9181 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     9241 acgacggcca gtgccaagct g
//
