LOCUS       V041150                 7698 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041150
VERSION     V041150
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2378
                     /label="Yes1(NM_009535)"
                     /note="Yes1(NM_009535)"
                     /gene="Yes1"
     CDS             2385..2450
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2474..3062
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3128..3176
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3378..3806
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3820..4149
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4216..5010
                     /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    5186..5307
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5356..5372)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5380..5396
                     /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(5404..5434)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5449..5470
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5758..6346)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6517..7377)
                     /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(7378..7482)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggg ctgcattaaa agtaaagaaa
      781 acaaaagtcc agccataaaa tacacaccgg aaaatcttac agagcctgta agcccaagtg
      841 ccagtcatta tggagtggaa catgctacag ttgccccgac ctcttccaca aagggagcat
      901 cagttaattt taacagtctt tccatgacac cctttggagg gtcctcaggg gtgactcctt
      961 ttggaggagc gtcttcctca ttctcagtgg tgtcaagttc atatcctaca ggtttaacag
     1021 gtggtgtcac tatatttgtg gccttgtatg attatgaagc tagaactaca gaagaccttt
     1081 cctttaagaa gggtgaacga tttcaaataa ttaacaatac ggaaggagac tggtgggaag
     1141 caagatcaat tgctaccgga aagagtggtt atatccctag caattacgta gtgcctgcag
     1201 attccattca ggcagaagaa tggtattttg gcaaaatggg gagaaaagat gcggaaagat
     1261 tacttctgaa tcctgggaat cagcgaggta ttttcttagt aagagaaagt gaaactacta
     1321 aaggtgctta ctccctctca atccgtgatt gggatgaggt gaggggtgac aatgtgaagc
     1381 attacaagat cagaaaactt gacaatggtg gctactacat cacgaccaga gctcagtttg
     1441 atacactgca gaagctggtg aagcactaca cagaacatgc tgatggatta tgccacaagt
     1501 taacaactgt gtgtcctact gtgaaacccc agactcaagg tctggcaaaa gatgcttggg
     1561 aaatccctcg agaatcattg cgactagagg tgaaactagg tcaaggatgc tttggggaag
     1621 tgtggatggg aacatggaat ggaactacaa aagtagcaat caaaacacta aagccaggta
     1681 caatgatgcc agaagcattc cttcaagaag ctcagataat gaaaaagcta agacacgata
     1741 aacttgttcc actctatgca gttgtttctg aagagcccat ttatattgtc accgagttta
     1801 tgtcaaaagg aagcttgtta gatttcctta aagaaggaga tggaaagtat ttgaagcttc
     1861 cacagctggt tgatatggct gctcagatcg ctgatggcat ggcgtatatt gaaagaatga
     1921 actatattca ccgagatctc cgagctgcta atattcttgt aggagaaaat cttatatgca
     1981 aaatagcaga ttttggctta gcaagattaa ttgaagacaa tgaatacacg gcaagacaag
     2041 gtgcaaaatt tccaatcaag tggacagctc ctgaggctgc tctgtatggt cgatttacaa
     2101 taaagtcaga tgtgtggtca tttggaattc tacagacaga gctggtaaca aaaggaagag
     2161 tgccatatcc aggtatggta aaccgtgaag tattggaaca agtagagcgg ggatacagaa
     2221 tgccttgccc ccagggctgt cccgaatccc tccatgaatt gatgaatctt tgctggaaga
     2281 aggatcctga tgaaagacca acatttgaat atattcagtc cttcttggaa gactacttca
     2341 ctgctacaga gccacagtac caaccaggag aaaatttagg atccgactac aaagaccatg
     2401 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc
     2461 ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     2521 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     2581 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     2641 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     2701 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     2761 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     2821 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     2881 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     2941 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3001 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     3061 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     3121 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     3181 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     3241 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     3301 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     3361 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3421 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3481 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     3541 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3601 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3661 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3721 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     3781 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3841 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3901 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3961 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4021 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4081 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4141 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     4201 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4261 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4321 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4381 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4441 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4501 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4561 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4621 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4681 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4741 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4801 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4861 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4921 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4981 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     5041 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5101 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5161 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5221 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5281 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5341 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5401 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5461 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5521 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5581 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5641 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5701 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5761 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5821 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5881 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5941 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6001 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6061 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6121 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6181 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6241 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6301 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6361 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6421 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6481 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6541 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6601 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6661 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6721 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6781 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6841 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6901 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6961 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7021 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7081 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7141 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7201 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7261 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7321 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7381 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7441 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7501 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
     7561 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     7621 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     7681 atgaagaatc tgcttagg
//
