LOCUS       V039359                10141 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039359
VERSION     V039359
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..5190
                     /label="Ireb2(NM_022655)"
                     /note="Ireb2(NM_022655)"
                     /gene="Ireb2"
     CDS             5197..5262
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        5306..5517
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5530..5798
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5831..6229
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     misc_feature    6236..6824
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6898..7131
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7203..7324
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7343..7478
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7516..7532)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7540..7556
                     /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(7564..7594)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7609..7630
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7918..8506)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8677..9537)
                     /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(9538..9642)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     10116..10132
                     /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 catggactcc ccaagtgcag gatacacctt tgagtacctt
     2341 attgaaacat taaatggcaa ttcacagaag aagttcttca atgtacctaa acttggcggc
     2401 accaagtatg atattctgcc ttactcaata cgggtcctat tggaagctgc tgtacgaaat
     2461 tgtgatggct ttttaatgaa aaaggaagat gttatgaaca ttttggactg gaaaaccaaa
     2521 cagagcaatg ttgaagtgcc ctttttccct gcccgtgttg ttcttcaaga tttcactgga
     2581 ataccggcaa tggtggattt tgctgctatg agggaggcag tgaaaactct tggaggtgat
     2641 cctaagaaag tccaccctgc ctgtccgaca gatctcacag tggaccactc tttacagatt
     2701 gacttcagta aatgtgcaat acagaatgca ccaaatcctg gaggtggtga cctacagaaa
     2761 gcaggaaagc tctctccact taaagtgcag tctaagaagc ttccatgtcg aggccagact
     2821 acctgccggg gatcgtgtga ttctggagag ctaagccgga actcaggaac attttcttca
     2881 cagattgaga atacacctgt cttgtgtcct tttcatttgc aaccagtgcc tgaacctgag
     2941 acagtgttaa aaaatcaaga agtagaattt ggcagaaatc gagagaggct tcagtttttc
     3001 aagtggagct caggagcttt taagaatgtg gcagtcatcc cgcctggaac tggaatggct
     3061 catcaagtga acttagaata tttatcaaga gtagtttttg aagaaacgga cctgctcttc
     3121 ccagacagtg tagttggcac agattctcat ataaccatgg tgaacggatt ggggattctt
     3181 gggtggggag ttggtggcat tgagacagag gcagttatgc ttggcctgcc agttactctt
     3241 actttaccag aggtggttgg atgtgagcta actgggtcat ccaatgcttt tgttacatcc
     3301 atagatattg tccttggcat tacaaagcac ctcaggcaag taggcgtggc tggaaagttt
     3361 gttgagttct ttggaagtgg agtttcacaa ttatctattg tcgatagaac taccatagca
     3421 aacatgtgtc cagaatatgg tgctatcctc agctttttcc ctgttgacaa tgtgacacta
     3481 cgacatttag aacatacagg ttttgacaaa accaaacttg agtcaatgga aaaatacctt
     3541 aaagctgtga aattgtttcg aaatgatgag aattcttcag aacctgaata ttctcaggtg
     3601 atacagatta atctgaattc aatagttgca tctgtcagtg gtccaaaaag gcctcaggat
     3661 agggttgctg taacagatat gaaaagtgat tttcaggctt gcttaaatga aaaggttgga
     3721 tttaaaggct tccaagttgc agcagaaaaa caaagtgaca ctgtctctgt tcgttatgat
     3781 ggaagtgagt ataagctgtc ccatggatcc gtggtcattg ctgcagttat cagttgtacc
     3841 aataattgca atccatctgt gatgcttgct gcaggtcttc tggctaaaaa ggctgttgaa
     3901 attggtctac gagttaaacc ttatatacga acaagtttgt ctccaggcag tgggatggtt
     3961 acacattacc tcagttcaag tggagtgcta ccctatctta gcaagctagg gtttgacata
     4021 gttggctatg gatgttcaac atgtgtagga aatacagcac ctttatcaga agcagttttg
     4081 aatgcagtaa aacagggtga tttggttacc tgtggagttt tatctggaaa caaacatttc
     4141 gaaggccgtc tctgtgattg tgtccgtgcc aattatctcg cctctccgcc cttagtggtg
     4201 gcttatgcca tagcaggcac agtgaatata gacttccaga cagagccttt aggtactgac
     4261 tctacgggca aggaaattta cctgcatgac atttggccta gtcgagaaga agttcatcag
     4321 atggaggaag aacatgttat attgtcaatg tttaaaacac taaaagagaa agtagagatg
     4381 ggaaataaac ggtggaattc gttagaagct ccagactccg tgctgtttcc atgggatgtg
     4441 aagtctactt acatcagatg cccttcattt tttgataagc ttaccaaaga gccagctgca
     4501 tcccagccta ttgagaatgc ccatgtcctg ttgtatttgg gagactctgt cactacagat
     4561 catatatcac ccgctggaag tattgctagg agtagtgctg ctgctaagta tttgacaaac
     4621 agaggcctta ctccccgaga gttcaactct tatggagctc gaagaggtaa tgatgctgtg
     4681 atgacaagag gcacatttgc aaacatcaag ctttttaata agtttattgg gaagccagct
     4741 cccaaaacaa ttcattttcc atcgggacaa acgctcgatg tatttgaagc tgcagaactg
     4801 taccagaaag aaggtatccc actgataatt ttagcaggaa aaaaatatgg ttcaggaaat
     4861 tcaagagatt gggctgccaa gggaccctat ttgctgggtg taaaagctgt tttggctgag
     4921 agctatgaaa aaatacacaa agatcatttg attggaattg gtatagcacc acttgaattc
     4981 ctcccaggag aaaatgcaga ttccttgggc ctctctggca gagaagtatt ttctttatca
     5041 tttcctgaag aactctttcc tggaattaca ttaaatataa agacaagcac tggaaaagag
     5101 ttcagcgtga ttgcatcatt tgcaaatgac gtggagataa ctttgtacaa acatggaggc
     5161 ttattaaact ttgtggctcg aaaattctta ggatccgact acaaagacca tgacggtgat
     5221 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc
     5281 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     5341 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg
     5401 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     5461 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg
     5521 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat
     5581 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc
     5641 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg
     5701 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac
     5761 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg
     5821 ctaggccacc atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac
     5881 ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc
     5941 tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc
     6001 agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt
     6061 cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct
     6121 tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc
     6181 agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag tcgacaatca
     6241 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     6301 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     6361 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     6421 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     6481 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     6541 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     6601 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     6661 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     6721 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     6781 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac
     6841 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     6901 aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact gggtctctct
     6961 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     7021 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     7081 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     7141 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     7201 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     7261 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     7321 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg cccattctcc
     7381 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     7441 gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca gagacggccc
     7501 aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7561 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     7621 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7681 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     7741 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7801 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7861 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7921 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7981 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     8041 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     8101 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     8161 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     8221 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     8281 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     8341 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     8401 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     8461 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     8521 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     8581 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     8641 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     8701 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     8761 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     8821 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     8881 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8941 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     9001 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     9061 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     9121 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     9181 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     9241 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     9301 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     9361 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     9421 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     9481 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     9541 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     9601 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     9661 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     9721 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     9781 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     9841 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     9901 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     9961 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
    10021 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
    10081 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct
    10141 g
//
