LOCUS       V032059                 8977 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032059
VERSION     V032059
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..3831
                     /label="Kdm4d(NM_173433)"
                     /note="Kdm4d(NM_173433)"
                     /gene="Kdm4d"
     CDS             3838..3903
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3947..4158
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4171..4439
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4466..5065
                     /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    5072..5660
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5734..5967
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6039..6160
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6179..6314
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6352..6368)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6376..6392
                     /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(6400..6430)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6445..6466
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6754..7342)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7513..8373)
                     /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(8374..8478)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8952..8968
                     /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 catgaagacg aagtccacat gtgctcagaa tccaaattgc
     2341 agcataatga tatttcgtcc aaccaaagaa gagtttaatg attttgacaa atacattgct
     2401 tacatggagt cccaaggggc acaccgagct ggactggcca aggtcatccc accaaaagaa
     2461 tggagggcca ggcagtctta tgacaatatc agcaacatct taatagcaac tcccctgcag
     2521 caagtggtct ctgggcaggc aggcgtgttc acccaatacc ataagaagaa gaaaggcatg
     2581 acagtggggg agtaccgtga gctggccaac agcaaaaagt accagacccc gccacacctg
     2641 gattttgaag atttggagcg aaaatactgg aagaatcgcc tgtatgagtc accgatttat
     2701 ggtgctgacg tcagcggctc cctgtttgat gggaagactc aacagtggaa tgtgggtcac
     2761 ctgggaacaa ttcaagacct attggaacag gaatgtggca tagtgattga gggcgtcaac
     2821 acgccctacc tgtactttgg catgtggaag accacctttg cgtggcacac ggaggacatg
     2881 gacctgtaca gtatcaacta cctacacttt gggcagccca agacctggta tgctgtgccc
     2941 cctgagcatg gcaggcgcct ggagcgcctg gccagggaac tcttccctgg cagctcccag
     3001 ggctgccagg ccttcctgag gcacaaggtg gcgctcatct cgcccactgt gcttaaggag
     3061 aatggcatcc cctttggtcg catcacccag gaggctgggg agttcatggt cacctttccc
     3121 tatggctacc acgcgggctt caaccatggc ttcaactgcg cagaggccat caattttgcc
     3181 accccaaggt ggattgacta tggcaaggtg gcatctcagt gcagctgtgg ggaggccagg
     3241 gtgagcttct ccatggacgc ctttgtgagg atcttgcagc ctgagcgata tgaactgtgg
     3301 aaacgtggcc aagatcaggc agttgtggac cacacagaga ctatggtgtc taccagtcag
     3361 gagctcacca cccggcgggt gaccaaagca ccaagaaaaa cttggggctt gaagcgtctc
     3421 cggcttcgcc aggtctcacg atctcttctg cctatagcca cggtaagtaa cgttccttgc
     3481 aacatgcagg tgtgccacac ctccaggcaa ccatcagatg tgaaaggtga tgatgtccag
     3541 aagtctgact cagccagagc ctcaccacat cctctgagtc tgccttcttc tggtcacatg
     3601 tcaactagaa gatgtagtct tggtcgtcgt ccttgtgaac taggagctca ggagtcctcc
     3661 aatggagctc cagtcaagag gcaacttcca gcaggcagag atgacacaag ccccagtcca
     3721 gagcttcagc cccaggctgt gagtggagac ttaatagtcg actcaggact tgtgaaccct
     3781 ggcccacagc atcttatgac agcttctgaa gggggattga cctccgaccc cggatccgac
     3841 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac
     3901 aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca
     3961 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga
     4021 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     4081 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     4141 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc
     4201 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt
     4261 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc
     4321 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg
     4381 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc
     4441 aagctgtgac cggcgcctac gctagatgac cgagtacaag cccacggtgc gcctcgccac
     4501 ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     4561 cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     4621 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     4681 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     4741 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     4801 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     4861 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     4921 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     4981 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     5041 catgacccgc aagcccggtg cctgagtcga caatcaacct ctggattaca aaatttgtga
     5101 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     5161 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     5221 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     5281 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     5341 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     5401 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     5461 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     5521 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     5581 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     5641 cctttgggcc gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag
     5701 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     5761 aataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     5821 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     5881 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     5941 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     6001 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     6061 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6121 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     6181 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     6241 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     6301 cttttttgga ggcctagact tttgcagaga cggcccaaat tcgtaatcat ggtcatagct
     6361 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6421 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6481 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6541 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6601 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6661 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6721 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6781 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6841 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6901 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6961 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7021 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7081 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7141 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     7201 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7261 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7321 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7381 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7441 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7501 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7561 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7621 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7681 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7741 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7801 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7861 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7921 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7981 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8041 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8101 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8161 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8221 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8281 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8341 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8401 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8461 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     8521 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     8581 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     8641 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     8701 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     8761 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     8821 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     8881 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     8941 gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
