LOCUS       V038246                 7887 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038246
VERSION     V038246
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..2567
                     /label="PTGS2(NM_000963)"
                     /note="PTGS2(NM_000963)"
                     /gene="PTGS2"
     CDS             2574..2639
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2663..3251
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3317..3365
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3567..3995
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4009..4338
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4405..5199
                     /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    5375..5496
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5545..5561)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5569..5585
                     /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(5593..5623)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5638..5659
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5947..6535)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6706..7566)
                     /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(7567..7671)
                     /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 ccaccatgct cgcccgcgcc ctgctgctgt
      781 gcgcggtcct ggcgctcagc catacagcaa atccttgctg ttcccaccca tgtcaaaacc
      841 gaggtgtatg tatgagtgtg ggatttgacc agtataagtg cgattgtacc cggacaggat
      901 tctatggaga aaactgctca acaccggaat ttttgacaag aataaaatta tttctgaaac
      961 ccactccaaa cacagtgcac tacatactta cccacttcaa gggattttgg aacgttgtga
     1021 ataacattcc cttccttcga aatgcaatta tgagttatgt gttgacatcc agatcacatt
     1081 tgattgacag tccaccaact tacaatgctg actatggcta caaaagctgg gaagccttct
     1141 ctaacctctc ctattatact agagcccttc ctcctgtgcc tgatgattgc ccgactccct
     1201 tgggtgtcaa aggtaaaaag cagcttcctg attcaaatga gattgtggaa aaattgcttc
     1261 taagaagaaa gttcatccct gatccccagg gctcaaacat gatgtttgca ttctttgccc
     1321 agcacttcac gcatcagttt ttcaagacag atcataagcg agggccagct ttcaccaacg
     1381 ggctgggcca tggggtggac ttaaatcata tttacggtga aactctggct agacagcgta
     1441 aactgcgcct tttcaaggat ggaaaaatga aatatcagat aattgatgga gagatgtatc
     1501 ctcccacagt caaagatact caggcagaga tgatctaccc tcctcaagtc cctgagcatc
     1561 tacggtttgc tgtggggcag gaggtctttg gtctggtgcc tggtctgatg atgtatgcca
     1621 caatctggct gcgggaacac aacagagtat gcgatgtgct taaacaggag catcctgaat
     1681 ggggtgatga gcagttgttc cagacaagca ggctaatact gataggagag actattaaga
     1741 ttgtgattga agattatgtg caacacttga gtggctatca cttcaaactg aaatttgacc
     1801 cagaactact tttcaacaaa caattccagt accaaaatcg tattgctgct gaatttaaca
     1861 ccctctatca ctggcatccc cttctgcctg acacctttca aattcatgac cagaaataca
     1921 actatcaaca gtttatctac aacaactcta tattgctgga acatggaatt acccagtttg
     1981 ttgaatcatt caccaggcaa attgctggca gggttgctgg tggtaggaat gttccacccg
     2041 cagtacagaa agtatcacag gcttccattg accagagcag gcagatgaaa taccagtctt
     2101 ttaatgagta ccgcaaacgc tttatgctga agccctatga atcatttgaa gaacttacag
     2161 gagaaaagga aatgtctgca gagttggaag cactctatgg tgacatcgat gctgtggagc
     2221 tgtatcctgc ccttctggta gaaaagcctc ggccagatgc catctttggt gaaaccatgg
     2281 tagaagttgg agcaccattc tccttgaaag gacttatggg taatgttata tgttctcctg
     2341 cctactggaa gccaagcact tttggtggag aagtgggttt tcaaatcatc aacactgcct
     2401 caattcagtc tctcatctgc aataacgtga agggctgtcc ctttacttca ttcagtgttc
     2461 cagatccaga gctcattaaa acagtcacca tcaatgcaag ttcttcccgc tccggactag
     2521 atgatatcaa tcccacagta ctactaaaag aacgttcgac tgaactggga tccgactaca
     2581 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     2641 agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     2701 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2761 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2821 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2881 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2941 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3001 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     3061 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3121 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3181 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3241 cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa
     3301 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt
     3361 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     3421 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     3481 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg
     3541 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3601 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3661 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3721 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3781 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3841 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3901 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     3961 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4021 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4081 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4141 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4201 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4261 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4321 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4381 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4441 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4501 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4561 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4621 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4681 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4741 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4801 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4861 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4921 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4981 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5041 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5101 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5161 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     5221 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5281 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5341 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5401 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5461 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5521 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5581 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5641 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5701 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5761 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5821 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5881 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5941 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6001 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6061 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6121 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6181 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6241 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6301 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6361 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6421 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6481 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6541 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6601 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6661 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6721 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6781 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6841 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6901 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6961 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7021 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7081 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7141 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7201 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7261 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7321 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7381 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7441 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7501 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7561 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7621 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7681 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact
     7741 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     7801 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     7861 gacaattgca tgaagaatct gcttagg
//
