LOCUS       V041202                 7411 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041202
VERSION     V041202
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             706..2067
                     /label="GABPA(NM_002040)"
                     /note="GABPA(NM_002040)"
                     /gene="GABPA"
     CDS             2074..2163
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2187..2775
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2841..2889
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3091..3519
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3533..3862
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3929..4723
                     /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    4899..5020
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5069..5085)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5093..5109
                     /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(5117..5147)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5162..5183
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5471..6059)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6230..7090)
                     /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(7091..7195)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatgac taaaagagaa
      721 gcagaggagc tgatagaaat tgagattgat ggaacagaga aagcagagtg cacagaagaa
      781 agcattgtag aacaaaccta cgcgccagct gaatgtgtaa gccaggccat agacatcaat
      841 gaaccaatag gcaatttaaa gaaactgcta gaaccaagac tacagtgttc tttggatgct
      901 catgaaattt gtctgcaaga tatccagctg gatccagaac gaagtttatt tgaccaagga
      961 gtaaaaacag atggaactgt acagcttagt gtacaggtaa tttcttacca aggaattgaa
     1021 ccaaagttaa acatccttga aattgttaaa cctgcggaca ctgttgaggt tgttattgat
     1081 ccagatgccc accatgctga atcagaagca catcttgttg aagaagctca agtgataact
     1141 cttgatggca caaaacacat cacaaccatt tcagatgaaa cttcagaaca agtgacaaga
     1201 tgggctgctg cactggaagg ctataggaaa gaacaagaac gccttgggat accctatgat
     1261 cccatacagt ggtccacaga ccaagtcctg cattgggtgg tttgggtaat gaaggaattc
     1321 agcatgaccg atatagacct caccacactc aacatttcgg ggagagaatt atgtagtctc
     1381 aaccaagaag atttttttca gcgggttcct cggggagaaa ttctctggag tcatctggaa
     1441 cttctccgaa aatatgtatt ggcaagtcaa gaacaacaga tgaatgaaat agttacaatt
     1501 gatcaacctg tgcaaattat tccagcatca gtgcaatctg ctacacctac taccattaaa
     1561 gttataaata gtagtgcgaa agcagccaaa gtacaaagag cgccgaggat ttcaggagaa
     1621 gatagaagct cacctgggaa cagaacagga aacaatggcc aaatccaact atggcagttt
     1681 ttgctagaac ttcttactga taaggacgct cgagactgca tttcttgggt tggtgatgaa
     1741 ggtgaattta agctaaatca gcctgaactg gttgcacaga aatggggaca gcgtaaaaat
     1801 aagcctacga tgaactatga gaaactcagt cgtgcattaa gatattatta cgatggggac
     1861 atgatttgta aagttcaagg caagagattt gtgtacaagt ttgtctgtga cttgaagact
     1921 cttattggat acagtgcagc ggagttgaac cgtttggtca cagaatgtga acagaagaaa
     1981 cttgcaaaga tgcagctcca tggaattgcc cagccagtca cagcagtagc tctggctact
     2041 gcttctctgc aaacggaaaa ggataatctc gaggaacaaa aactaatatc agaggaagat
     2101 ttggaacaga aactaatcag tgaagaagat ttagagcaaa agttaatttc cgaggaggac
     2161 ttatagtgag ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     2221 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     2281 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     2341 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     2401 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     2461 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     2521 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     2581 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     2641 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     2701 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     2761 gggccgcctc cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc
     2821 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt
     2881 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     2941 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     3001 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc
     3061 tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     3121 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3181 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3241 atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3301 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3361 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3421 accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt
     3481 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3541 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3601 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3661 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3721 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3781 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     3841 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     3901 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3961 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4021 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4081 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4141 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4201 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4261 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4321 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4381 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4441 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4501 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4561 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4621 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4681 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4741 gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4801 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4861 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4921 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4981 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5041 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5101 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5161 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5221 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5281 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5341 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5401 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5461 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5521 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5581 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5641 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     5701 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5761 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5821 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5881 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     5941 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6001 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6061 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6121 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6181 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6241 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6301 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6361 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6421 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6481 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6541 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6601 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6661 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6721 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6781 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6841 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6901 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6961 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7021 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7081 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7141 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7201 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc
     7261 gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     7321 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     7381 gaccgacaat tgcatgaaga atctgcttag g
//
