LOCUS       V019882                 8177 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019882
VERSION     V019882
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..2857
                     /label="MCCC1(NM_020166)"
                     /note="MCCC1(NM_020166)"
                     /gene="MCCC1"
     CDS             2864..2929
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2953..3541
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3607..3655
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3857..4285
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4299..4628
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4695..5489
                     /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    5665..5786
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5835..5851)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5859..5875
                     /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(5883..5913)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5928..5949
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6237..6825)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6996..7856)
                     /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(7857..7961)
                     /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 ccatggcggc ggcctctgcg gtgtcggtgc tgctggtggc
      721 ggcggagagg aaccggtggc atcgtctccc gagcctgctc ctgccgccga ggacatgggt
      781 gtggaggcaa agaaccatga agtacacaac agccacagga agaaacatta ccaaggtcct
      841 cattgcaaac agaggagaaa ttgcctgcag ggtgatgcgc acagccaaaa aactgggtgt
      901 acagactgtg gcggtttata gtgaggctga cagaaattcc atgcatgtag atatggcaga
      961 tgaagcatat tccatcggcc ccgctccctc ccagcagagc tacctatcta tggagaaaat
     1021 cattcaagtg gccaagacct ctgctgcaca ggctatccat ccaggatgcg gttttctctc
     1081 agaaaacatg gaatttgctg aactttgtaa gcaagaagga attattttta taggccctcc
     1141 tccatctgca attagagaca tgggtataaa gagcacatcc aaatccataa tggctgctgc
     1201 tggagtacct gttgtggagg gttatcatgg tgaggaccaa tcagaccagt gcctgaagga
     1261 acacgccagg agaattggct atcctgtcat gattaaagcc gtccggggtg gaggaggaaa
     1321 aggaatgagg attgttagat cagaacaaga atttcaagaa cagttagagt cagcacggag
     1381 agaagctaag aagtctttca atgatgatgc tatgctgatc gagaagtttg tagacacacc
     1441 gaggcatgta gaagtccagg tgtttggtga tcaccatggc aatgctgtgt acttgtttga
     1501 aagagactgt agtgtgcaga ggcgacatca gaagatcatt gaggaggccc cagcgcctgg
     1561 tattaaatct gaagtaagaa aaaagctggg agaagctgca gtcagagctg ctaaagctgt
     1621 aaattatgtt ggagcaggga ctgtggagtt tattatggac tcaaaacata atttctgttt
     1681 catggagatg aatacaaggc tgcaagtgga acatcctgtt actgagatga tcacaggaac
     1741 tgacttggtg gagtggcagc ttagaattgc agcaggagag aagattcctt tgagccagga
     1801 agaaataact ctgcagggcc atgccttcga agctagaata tatgcagaag atcctagcaa
     1861 taacttcatg cctgtggcag gcccattagt gcacctctct actcctcgag cagacccttc
     1921 caccaggatt gaaactggag tacggcaagg agacgaagtt tccgtgcatt atgaccccat
     1981 gattgcgaag ctggtcgtgt gggcagcaga tcgccaggcg gcattgacaa aactgaggta
     2041 cagccttcgt cagtacaata ttgttggact gcacaccaac attgacttct tactcaacct
     2101 gtctggccac ccagagtttg aagctgggaa cgtgcacact gatttcatcc ctcaacacca
     2161 caaacagttg ttgctcagtc ggaaggctgc agccaaagag tctttatgcc aggcagccct
     2221 gggtctcatc ctcaaggaga aagccatgac cgacactttc actcttcagg cacatgatca
     2281 attctctcca ttttcgtcta gcagtggaag aagactgaat atctcgtata ccagaaacat
     2341 gactcttaaa gatggtaaaa acaatgtagc catagctgta acgtataacc atgatgggtc
     2401 ttatagcatg cagattgaag ataaaacttt ccaagtcctt ggtaatcttt acagcgaggg
     2461 agactgcact tacctgaaat gttctgttaa tggagttgct agtaaagcga agctgattat
     2521 cctggaaaac actatttacc tattttccaa ggaaggaagt attgagattg acattccagt
     2581 ccccaaatac ttatcttctg tgagctcaca agaaactcag ggcggcccct tagctcctat
     2641 gactggaacc attgaaaagg tgtttgtcaa agctggagac aaagtgaaag cgggagattc
     2701 cctcatggtt atgatcgcca tgaagatgga gcataccata aagtctccaa aggatggcac
     2761 agtaaagaaa gtgttctaca gagaaggtgc tcaggccaac agacacactc ctttagtcga
     2821 gtttgaggag gaagaatcag acaaaaggga atcggaactc gaggactaca aagaccatga
     2881 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc
     2941 cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     3001 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     3061 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     3121 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     3181 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     3241 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     3301 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     3361 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     3421 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     3481 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     3541 cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     3601 ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa
     3661 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     3721 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     3781 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc
     3841 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3901 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3961 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct
     4021 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4081 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4141 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4201 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg
     4261 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4321 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4381 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4441 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4501 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4561 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4621 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4681 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4741 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4801 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4861 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4921 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4981 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5041 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5101 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5161 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5221 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5281 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5341 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5401 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5461 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac
     5521 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5581 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5641 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     5701 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5761 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5821 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5881 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5941 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6001 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     6061 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6121 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6181 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6241 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6301 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6361 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6421 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6481 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6541 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6601 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6661 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6721 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6781 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6841 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6901 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6961 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7021 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     7081 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     7141 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7201 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7261 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7321 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7381 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7441 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7501 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7561 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7621 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7681 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7741 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7801 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7861 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7921 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7981 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa
     8041 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     8101 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     8161 tgaagaatct gcttagg
//
