LOCUS       V021133                 9575 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021133
VERSION     V021133
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             683..4255
                     /label="ZNF91(NM_003430)"
                     /note="ZNF91(NM_003430)"
                     /gene="ZNF91"
     CDS             4262..4327
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4351..4939
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5005..5053
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5255..5683
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5697..6026
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6093..6887
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    7063..7184
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7233..7249)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7257..7273
                     /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(7281..7311)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7326..7347
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7635..8223)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8394..9254)
                     /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(9255..9359)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgccagg aacccctgga agcctagaaa tgggactgtt
      721 gacatttagg gatgtggcca tagaattctc tccggaggag tggcaatgtc tggacactgc
      781 acagcagaat ttatatagga atgtgatgtt agagaactac agaaacctgg ccttcctggg
      841 tattgctctc tctaagccag acctgattac ttatctggag caaggaaaag agccctggaa
      901 tatgaagcaa catgagatgg tggatgaacc cacaggtata tgtcctcatt ttcctcaaga
      961 cttttggcca gagcagagca tggaagattc ttttcaaaaa gtattactga gaaaatatga
     1021 aaaatgtgga catgagaatt tacagttaag aaaaggttgt aaaagtgtgg atgagtgtaa
     1081 ggtgcacaaa gaaggttata ataaacttaa ccagtgtctc acaactgccc agagcaaagt
     1141 atttcaatgt gggaaatatt tgaaagtctt ctataaattt ttaaattcaa acagacatac
     1201 gataagacat actggaaaga aatgcttcaa atgtaaaaaa tgtgtcaagt cattttgcat
     1261 ccgtttacac aaaacccaac ataaatgcgt ttatattaca gagaagtcct gtaaatgtaa
     1321 agaatgtgaa aaaacctttc attggtcctc aacccttact aatcataagg aaattcatac
     1381 tgaagataaa ccctacaaat gtgaagaatg tggcaaagct tttaagcagc tctcaaccct
     1441 tactacacat aaaataatct gtgctaaaga gaaaatctac aagtgtgaag aatgtggcaa
     1501 agcatttcta tggtcctcaa ccctaactag acataagagg atacacactg gagagaaacc
     1561 ctacaaatgt gaagaatgtg gcaaagcttt tagccattct tcaacccttg ctaaacataa
     1621 gagaattcat actggagaga aaccctacaa atgtgaagaa tgtggcaaag cttttagccg
     1681 ttcttcaacc cttgctaaac ataagagaat tcatactgga gagaaaccct acaaatgtaa
     1741 agaatgtggc aaagctttta gcaattcctc aacccttgct aatcataaga taactcatac
     1801 tgaagagaaa ccctacaaat gtaaagaatg tgacaaagct tttaagcgac tctcaaccct
     1861 tactaaacat aaaataatac atgctggaga gaaactctac aaatgtgaag aatgtggcaa
     1921 agcttttaat cgatcttcaa atcttactat acataagttt attcatactg gagagaaacc
     1981 ttacaagtgt gaagaatgtg gcaaagcatt taactggtcc tcaagcctta ctaaacataa
     2041 aagatttcat actagagaga aacccttcaa atgtaaagaa tgtggcaaag catttatatg
     2101 gtcttcaacc ctaactagac ataagaggat acacactgga gagaagccct acaaatgtga
     2161 agaatgtggc aaagctttta ggcaatcctc aacccttact aaacataaga taattcatac
     2221 tggagagaaa ccctacaaat ttgaagaatg tggcaaagct tttagacaat ccttaaccct
     2281 taataaacat aagataattc atagtagaga gaaaccctac aaatgtaaag aatgtggcaa
     2341 agcttttaag caattctcaa cccttactac acataaaata attcatgctg gaaagaaact
     2401 ctacaaatgt gaagaatgtg gcaaagcttt taatcattcc tcaagtcttt ctacacataa
     2461 gataattcat actggagaga agtcttacaa gtgtgaagaa tgtggcaaag catttctatg
     2521 gtcctcaacc ctaagaagac ataagaggat acacactgga gagaaaccct acaaatgtga
     2581 agaatgtggc aaagctttta gccattcttc agcccttgct aaacataaga gaattcatac
     2641 tggagagaaa ccctacaaat gtaaagaatg tggcaaagct tttagcaatt cctcaaccct
     2701 tgctaatcat aagataactc atactgaaga gaaaccctac aaatgtaaag aatgtgacaa
     2761 aacttttaag cgactctcaa cccttactaa acataaaata atacatgctg gagagaaact
     2821 ctacaaatgt gaagaatgtg gcaaagcttt taatcgatct tcaaatctta ctatacataa
     2881 gtttattcat actggagaga aaccttacaa gtgtgaagaa tgtggcaaag catttaactg
     2941 gtcctcaagc cttactaaac ataaaagaat tcatactaga gagaaaccct tcaaatgtaa
     3001 agaatgtggc aaagcattta tatggtcttc aaccctaact agacataaga ggatacacac
     3061 tggagagaag ccctacaaat gtgaagaatg tggcaaagct tttagccgtt cctcaaccct
     3121 tactaagcat aagacaattc atactggaga gaaaccctac aaatgtaaag aatgtggcaa
     3181 agcttttaag cactcctcag cccttgctaa acataaaata atacatgctg gagagaaact
     3241 ctacaaatgt gaggaatgtg gcaaagcttt taatcaatct tcaaatctta cgacacataa
     3301 gataattcat actaaagaga aaccttccaa gagtgaagaa tgtgacaaag catttatctg
     3361 gtcctcaacc cttactgaac ataagagaat tcataccaga gagaaaacct acaaatgtga
     3421 agaatgtggc aaagcattta gccagccttc acaccttact acacataaga ggatgcacac
     3481 tggagagaaa ccctacaaat gtgaagaatg tggcaaagct tttagccaat cctcaaccct
     3541 tactacacat aagataattc atactggaga gaaaccctac aaatgtgaag aatgtggcaa
     3601 agcttttagg aaatcttcaa ctcttactga acataagata attcatactg gagagaaacc
     3661 ctacaaatgt gaagaatgtg gcaaagcatt tagccaatcc tcaaccctaa ctagacatac
     3721 gaggatgcac actggagaga aaccatacaa atgtgaagaa tgtggcaaag cttttaatcg
     3781 atcctcaaag cttactacac ataagataat tcatactgga gagaaacctt acaagtgtga
     3841 agaatgtggc aaagcattta tatcatcctc aaccctaaat ggacataaga gaattcatac
     3901 tagagagaaa ccctacaaat gtgaagaatg tggcaaagca tttagccaat cttcaaccct
     3961 aactagacat aagaggttgc acaccggaga gaaaccctac aaatgtggag aatgtggcaa
     4021 agcctttaaa gagtcctcag ctcttactaa acataagata attcacactg gagagaaacc
     4081 ctacaaatgt gaaaaatgtg gcaaagcctt taaccagtct tcaatcctta ctaaccataa
     4141 gaaaattcat actatcacac ctgtaatccc actactttgg gaggccgagg cgggcggatc
     4201 acgaggtcag gagatggaga ccatcctggc taacacagtg aaaccccttc tctacctcga
     4261 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     4321 tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa
     4381 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     4441 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     4501 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     4561 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     4621 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     4681 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     4741 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     4801 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     4861 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     4921 ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa
     4981 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt
     5041 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     5101 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     5161 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca
     5221 gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     5281 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     5341 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     5401 ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     5461 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     5521 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     5581 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat
     5641 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     5701 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5761 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5821 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5881 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5941 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     6001 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     6061 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     6121 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     6181 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     6241 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     6301 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     6361 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     6421 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     6481 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     6541 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     6601 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     6661 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     6721 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     6781 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     6841 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     6901 gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     6961 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     7021 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     7081 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     7141 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     7201 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     7261 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     7321 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     7381 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     7441 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     7501 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     7561 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7621 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7681 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7741 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7801 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     7861 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7921 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7981 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     8041 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     8101 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     8161 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     8221 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     8281 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     8341 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     8401 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     8461 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     8521 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     8581 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     8641 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     8701 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     8761 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     8821 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8881 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8941 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     9001 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     9061 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     9121 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     9181 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     9241 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     9301 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     9361 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     9421 ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     9481 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     9541 gcttgaccga caattgcatg aagaatctgc ttagg
//
