LOCUS       V025187                 8827 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025187
VERSION     V025187
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"
     regulatory      655..664
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             664..729
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             739..3537
                     /label="PGR(NM_000926)"
                     /note="PGR(NM_000926)"
                     /gene="PGR"
     misc_feature    3603..4191
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4257..4305
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4507..4935
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4949..5278
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5345..6139
                     /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    6315..6436
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6485..6501)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6509..6525
                     /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(6533..6563)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6578..6599
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6887..7475)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7646..8506)
                     /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(8507..8611)
                     /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 ttaagccacc
      661 atggactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
      721 gatgacaagc ttggtaccat gactgagctg aaggcaaagg gtccccgggc tccccacgtg
      781 gcgggcggcc cgccctcccc cgaggtcgga tccccactgc tgtgtcgccc agccgcaggt
      841 ccgttcccgg ggagccagac ctcggacacc ttgcctgaag tttcggccat acctatctcc
      901 ctggacgggc tactcttccc tcggccctgc cagggacagg acccctccga cgaaaagacg
      961 caggaccagc agtcgctgtc ggacgtggag ggcgcatatt ccagagctga agctacaagg
     1021 ggtgctggag gcagcagttc tagtccccca gaaaaggaca gcggactgct ggacagtgtc
     1081 ttggacactc tgttggcgcc ctcaggtccc gggcagagcc aacccagccc tcccgcctgc
     1141 gaggtcacca gctcttggtg cctgtttggc cccgaacttc ccgaagatcc accggctgcc
     1201 cccgccaccc agcgggtgtt gtccccgctc atgagccggt ccgggtgcaa ggttggagac
     1261 agctccggga cggcagctgc ccataaagtg ctgccccggg gcctgtcacc agcccggcag
     1321 ctgctgctcc cggcctctga gagccctcac tggtccgggg ccccagtgaa gccgtctccg
     1381 caggccgctg cggtggaggt tgaggaggag gatggctctg agtccgagga gtctgcgggt
     1441 ccgcttctga agggcaaacc tcgggctctg ggtggcgcgg cggctggagg aggagccgcg
     1501 gctgtcccgc cgggggcggc agcaggaggc gtcgccctgg tccccaagga agattcccgc
     1561 ttctcagcgc ccagggtcgc cctggtggag caggacgcgc cgatggcgcc cgggcgctcc
     1621 ccgctggcca ccacggtgat ggatttcatc cacgtgccta tcctgcctct caatcacgcc
     1681 ttattggcag cccgcactcg gcagctgctg gaagacgaaa gttacgacgg cggggccggg
     1741 gctgccagcg cctttgcccc gccgcggagt tcaccctgtg cctcgtccac cccggtcgct
     1801 gtaggcgact tccccgactg cgcgtacccg cccgacgccg agcccaagga cgacgcgtac
     1861 cctctctata gcgacttcca gccgcccgct ctaaagataa aggaggagga ggaaggcgcg
     1921 gaggcctccg cgcgctcccc gcgttcctac cttgtggccg gtgccaaccc cgcagccttc
     1981 ccggatttcc cgttggggcc accgcccccg ctgccgccgc gagcgacccc atccagaccc
     2041 ggggaagcgg cggtgacggc cgcacccgcc agtgcctcag tctcgtctgc gtcctcctcg
     2101 gggtcgaccc tggagtgcat cctgtacaaa gcggagggcg cgccgcccca gcagggcccg
     2161 ttcgcgccgc cgccctgcaa ggcgccgggc gcgagcggct gcctgctccc gcgggacggc
     2221 ctgccctcca cctccgcctc tgccgccgcc gccggggcgg cccccgcgct ctaccctgca
     2281 ctcggcctca acgggctccc gcagctcggc taccaggccg ccgtgctcaa ggagggcctg
     2341 ccgcaggtct acccgcccta tctcaactac ctgaggccgg attcagaagc cagccagagc
     2401 ccacaataca gcttcgagtc attacctcag aagatttgtt taatctgtgg ggatgaagca
     2461 tcaggctgtc attatggtgt ccttacctgt gggagctgta aggtcttctt taagagggca
     2521 atggaagggc agcacaacta cttatgtgct ggaagaaatg actgcatcgt tgataaaatc
     2581 cgcagaaaaa actgcccagc atgtcgcctt agaaagtgct gtcaggctgg catggtcctt
     2641 ggaggtcgaa aatttaaaaa gttcaataaa gtcagagttg tgagagcact ggatgctgtt
     2701 gctctcccac agccagtggg cgttccaaat gaaagccaag ccctaagcca gagattcact
     2761 ttttcaccag gtcaagacat acagttgatt ccaccactga tcaacctgtt aatgagcatt
     2821 gaaccagatg tgatctatgc aggacatgac aacacaaaac ctgacacctc cagttctttg
     2881 ctgacaagtc ttaatcaact aggcgagagg caacttcttt cagtagtcaa gtggtctaaa
     2941 tcattgccag gttttcgaaa cttacatatt gatgaccaga taactctcat tcagtattct
     3001 tggatgagct taatggtgtt tggtctagga tggagatcct acaaacacgt cagtgggcag
     3061 atgctgtatt ttgcacctga tctaatacta aatgaacagc ggatgaaaga atcatcattc
     3121 tattcattat gccttaccat gtggcagatc ccacaggagt ttgtcaagct tcaagttagc
     3181 caagaagagt tcctctgtat gaaagtattg ttacttctta atacaattcc tttggaaggg
     3241 ctacgaagtc aaacccagtt tgaggagatg aggtcaagct acattagaga gctcatcaag
     3301 gcaattggtt tgaggcaaaa aggagttgtg tcgagctcac agcgtttcta tcaacttaca
     3361 aaacttcttg ataacttgca tgatcttgtc aaacaacttc atctgtactg cttgaataca
     3421 tttatccagt cccgggcact gagtgttgaa tttccagaaa tgatgtctga agttattgct
     3481 gcacaattac ccaagatatt ggcagggatg gtgaaacccc ttctctttca taaaaagtga
     3541 gaattctgca gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgatatctcg
     3601 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     3661 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     3721 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     3781 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     3841 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     3901 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     3961 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     4021 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     4081 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     4141 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg
     4201 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     4261 aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc
     4321 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     4381 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     4441 ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt
     4501 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4561 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4621 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc
     4681 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4741 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4801 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     4861 atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4921 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4981 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     5041 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5101 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     5161 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     5221 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     5281 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     5341 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     5401 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     5461 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     5521 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     5581 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     5641 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     5701 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     5761 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     5821 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     5881 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5941 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     6001 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     6061 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     6121 tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc
     6181 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     6241 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     6301 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     6361 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     6421 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     6481 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     6541 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     6601 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     6661 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     6721 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6781 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6841 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6901 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6961 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     7021 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     7081 aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7141 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7201 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7261 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7321 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7381 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7441 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7501 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7561 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7621 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7681 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7741 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7801 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7861 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7921 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7981 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     8041 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     8101 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     8161 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     8221 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     8281 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     8341 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     8401 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     8461 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     8521 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     8581 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     8641 tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga
     8701 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     8761 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     8821 gcttagg
//
