LOCUS       V042413                 9887 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042413
VERSION     V042413
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             2290..4542
                     /label="EZH2(NM_004456)"
                     /note="EZH2(NM_004456)"
                     /gene="EZH2"
     promoter        4656..4867
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4880..5148
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5181..5975
                     /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    5982..6570
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6644..6877
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6949..7070
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7089..7224
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7262..7278)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7286..7302
                     /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(7310..7340)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7355..7376
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7664..8252)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8423..9283)
                     /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(9284..9388)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9862..9878
                     /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 agagccacca tgggccagac tgggaagaaa tctgagaagg gaccagtttg ttggcggaag
     2341 cgtgtaaaat cagagtacat gcgactgaga cagctcaaga ggttcagacg agctgatgaa
     2401 gtaaagagta tgtttagttc caatcgtcag aaaattttgg aaagaacgga aatcttaaac
     2461 caagaatgga aacagcgaag gatacagcct gtgcacatcc tgacttctgt gagctcattg
     2521 cgcgggacta gggagtgttc ggtgaccagt gacttggatt ttccaacaca agtcatccca
     2581 ttaaagactc tgaatgcagt tgcttcagta cccataatgt attcttggtc tcccctacag
     2641 cagaatttta tggtggaaga tgaaactgtt ttacataaca ttccttatat gggagatgaa
     2701 gttttagatc aggatggtac tttcattgaa gaactaataa aaaattatga tgggaaagta
     2761 cacggggata gagaatgtgg gtttataaat gatgaaattt ttgtggagtt ggtgaatgcc
     2821 cttggtcaat ataatgatga tgacgatgat gatgatggag acgatcctga agaaagagaa
     2881 gaaaagcaga aagatctgga ggatcaccga gatgataaag aaagccgccc acctcggaaa
     2941 tttccttctg ataaaatttt tgaagccatt tcctcaatgt ttccagataa gggcacagca
     3001 gaagaactaa aggaaaaata taaagaactc accgaacagc agctcccagg cgcacttcct
     3061 cctgaatgta cccccaacat agatggacca aatgctaaat ctgttcagag agagcaaagc
     3121 ttacactcct ttcatacgct tttctgtagg cgatgtttta aatatgactg cttcctacat
     3181 cgtaagtgca attattcttt tcatgcaaca cccaacactt ataagcggaa gaacacagaa
     3241 acagctctag acaacaaacc ttgtggacca cagtgttacc agcatttgga gggagcaaag
     3301 gagtttgctg ctgctctcac cgctgagcgg ataaagaccc caccaaaacg tccaggaggc
     3361 cgcagaagag gacggcttcc caataacagt agcaggccca gcacccccac cattaatgtg
     3421 ctggaatcaa aggatacaga cagtgatagg gaagcaggga ctgaaacggg gggagagaac
     3481 aatgataaag aagaagaaga gaagaaagat gaaacttcga gctcctctga agcaaattct
     3541 cggtgtcaaa caccaataaa gatgaagcca aatattgaac ctcctgagaa tgtggagtgg
     3601 agtggtgctg aagcctcaat gtttagagtc ctcattggca cttactatga caatttctgt
     3661 gccattgcta ggttaattgg gaccaaaaca tgtagacagg tgtatgagtt tagagtcaaa
     3721 gaatctagca tcatagctcc agctcccgct gaggatgtgg atactcctcc aaggaaaaag
     3781 aagaggaaac accggttgtg ggctgcacac tgcagaaaga tacagctgaa aaaggacggc
     3841 tcctctaacc atgtttacaa ctatcaaccc tgtgatcatc cacggcagcc ttgtgacagt
     3901 tcgtgccctt gtgtgatagc acaaaatttt tgtgaaaagt tttgtcaatg tagttcagag
     3961 tgtcaaaacc gctttccggg atgccgctgc aaagcacagt gcaacaccaa gcagtgcccg
     4021 tgctacctgg ctgtccgaga gtgtgaccct gacctctgtc ttacttgtgg agccgctgac
     4081 cattgggaca gtaaaaatgt gtcctgcaag aactgcagta ttcagcgggg ctccaaaaag
     4141 catctattgc tggcaccatc tgacgtggca ggctggggga tttttatcaa agatcctgtg
     4201 cagaaaaatg aattcatctc agaatactgt ggagagatta tttctcaaga tgaagctgac
     4261 agaagaggga aagtgtatga taaatacatg tgcagctttc tgttcaactt gaacaatgat
     4321 tttgtggtgg atgcaacccg caagggtaac aaaattcgtt ttgcaaatca ttcggtaaat
     4381 ccaaactgct atgcaaaagt tatgatggtt aacggtgatc acaggatagg tatttttgcc
     4441 aagagagcca tccagactgg cgaagagctg ttttttgatt acagatacag ccaggctgat
     4501 gccctgaagt atgtcggcat cgaaagagaa atggaaatcc ctggtggcag tggtggtggg
     4561 agcggcgtga ctggatacag actttttgaa gagatcctgg aattctaagg atccgcggcc
     4621 gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     4681 ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg
     4741 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     4801 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     4861 aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg
     4921 aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg
     4981 aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc
     5041 gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc
     5101 tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc
     5161 ggcgcctacg ctaggccacc atggctgaac aagatggatt gcacgcaggt tctccggccg
     5221 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5281 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5341 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     5401 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5461 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5521 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5581 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5641 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5701 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5761 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5821 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5881 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5941 tcgccttcta tcgccttctt gacgagttct tctgagtcga caatcaacct ctggattaca
     6001 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     6061 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     6121 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     6181 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     6241 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     6301 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     6361 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     6421 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     6481 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     6541 gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat gacttacaag
     6601 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     6661 tcccaacgaa aataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     6721 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     6781 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     6841 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     6901 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     6961 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     7021 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     7081 ctctagctat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     7141 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     7201 agtgaggagg cttttttgga ggcctagact tttgcagaga cggcccaaat tcgtaatcat
     7261 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     7321 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     7381 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     7441 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     7501 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     7561 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     7621 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     7681 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     7741 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     7801 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     7861 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7921 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7981 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     8041 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     8101 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     8161 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     8221 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     8281 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     8341 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     8401 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     8461 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     8521 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     8581 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     8641 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     8701 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     8761 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     8821 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     8881 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     8941 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     9001 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     9061 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     9121 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     9181 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     9241 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     9301 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     9361 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     9421 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     9481 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     9541 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     9601 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     9661 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     9721 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     9781 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     9841 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
