LOCUS       V041523                 6887 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041523
VERSION     V041523
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..1534
                     /label="TPM1(NM_001018005)"
                     /note="TPM1(NM_001018005)"
                     /gene="TPM1"
     CDS             1574..1639
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1663..2251
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2317..2365
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2567..2995
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3009..3338
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3405..4199
                     /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    4375..4496
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4545..4561)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4569..4585
                     /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(4593..4623)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4638..4659
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4947..5535)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5706..6566)
                     /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(6567..6671)
                     /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 ccatggacgc catcaagaag aagatgcaga tgctgaagct
      721 cgacaaggag aacgccttgg atcgagctga gcaggcggag gccgacaaga aggcggcgga
      781 agacaggagc aagcagctgg aagatgagct ggtgtcactg caaaagaaac tcaagggcac
      841 cgaagatgaa ctggacaaat actctgaggc tctcaaagat gcccaggaga agctggagct
      901 ggcagagaaa aaggccaccg atgctgaagc cgacgtagct tctctgaaca gacgcatcca
      961 gctggttgag gaagagttgg atcgtgccca ggagcgtctg gcaacagctt tgcagaagct
     1021 ggaggaagct gagaaggcag cagatgagag tgagagaggc atgaaagtca ttgagagtcg
     1081 agcccaaaaa gatgaagaaa aaatggaaat tcaggagatc caactgaaag aggccaagca
     1141 cattgctgaa gatgccgacc gcaaatatga agaggtggcc cgtaagctgg tcatcattga
     1201 gagcgacctg gaacgtgcag aggagcgggc tgagctctca gaaggcaaat gtgccgagct
     1261 tgaagaagaa ttgaaaactg tgacgaacaa cttgaagtca ctggaggctc aggctgagaa
     1321 gtactcgcag aaggaagaca gatatgagga agagatcaag gtcctttccg acaagctgaa
     1381 ggaggctgag actcgggctg agtttgcgga gaggtcagta actaaattgg agaaaagcat
     1441 tgatgactta gaagacgagc tgtacgctca gaaactgaag tacaaagcca tcagcgagga
     1501 gctggaccac gctctcaacg atatgacttc catagaattc tgcagatatc cagcacagtg
     1561 gcggccgctc gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta
     1621 caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc tctggattac
     1681 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     1741 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     1801 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     1861 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     1921 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     1981 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     2041 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg
     2101 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     2161 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     2221 agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac
     2281 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     2341 cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     2401 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     2461 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     2521 cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc
     2581 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2641 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2701 ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac
     2761 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2821 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2881 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg
     2941 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3001 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3061 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3121 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3181 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3241 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3301 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3361 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3421 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3481 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3541 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3601 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3661 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3721 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3781 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3841 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3901 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     3961 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4021 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4081 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4141 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4201 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4261 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4321 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     4381 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4441 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4501 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     4561 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4621 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4681 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4741 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4801 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4861 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4921 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4981 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5041 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5101 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     5161 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5221 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5281 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5341 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     5401 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5461 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5521 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5581 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5641 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5701 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5761 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5821 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5881 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5941 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6001 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6061 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6121 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6181 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6241 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6301 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6361 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6421 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6481 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6541 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6601 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6661 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     6721 ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     6781 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     6841 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
