LOCUS       V019295                 7627 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019295
VERSION     V019295
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             928..1017
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     CDS             1051..3180
                     /label="NCL(NM_005381)"
                     /note="NCL(NM_005381)"
                     /gene="NCL"
     polyA_signal    3227..3451
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3497..3925
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3939..4268
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4335..5129
                     /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    5303..5424
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5473..5489)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5497..5513
                     /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(5521..5551)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5566..5587
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5875..6460)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6631..7491)
                     /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(7492..7596)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagccaccat ggcggcggaa caaaaactaa tatcagagga agatttggaa
      961 cagaaactaa tcagtgaaga agatttagag caaaagttaa tttccgagga ggacttatca
     1021 gcggaaaagc ttggtaccga gctcggatcc atggtgaagc tcgcgaaggc aggtaaaaat
     1081 caaggtgacc ccaagaaaat ggctcctcct ccaaaggagg tagaagaaga tagtgaagat
     1141 gaggaaatgt cagaagatga agaagatgat agcagtggag aagaggtcgt catacctcag
     1201 aagaaaggca agaaggctgc tgcaacctca gcaaagaagg tggtcgtttc cccaacaaaa
     1261 aaggttgcag ttgccacacc agccaagaaa gcagctgtca ctccaggcaa aaaggcagca
     1321 gcaacacctg ccaagaagac agttacacca gccaaagcag ttaccacacc tggcaagaag
     1381 ggagccacac caggcaaagc attggtagca actcctggta agaagggtgc tgccatccca
     1441 gccaaggggg caaagaatgg caagaatgcc aagaaggaag acagtgatga agaggaggat
     1501 gatgacagtg aggaggatga ggaggatgac gaggacgagg atgaggatga agatgaaatt
     1561 gaaccagcag cgatgaaagc agcagctgct gcccctgcct cagaggatga ggacgatgag
     1621 gatgacgaag atgatgagga tgacgatgac gatgaggaag atgactctga agaagaagct
     1681 atggagacta caccagccaa aggaaagaaa gctgcaaaag ttgttcctgt gaaagccaag
     1741 aacgtggctg aggatgaaga tgaagaagag gatgatgagg acgaggatga cgacgacgac
     1801 gaagatgatg aagatgatga tgatgaagat gatgaggagg aggaagaaga ggaggaggaa
     1861 gagcctgtca aagaagcacc tggaaaacga aagaaggaaa tggccaaaca gaaagcagct
     1921 cctgaagcca agaaacagaa agtggaaggc acagaaccga ctacggcttt caatctcttt
     1981 gttggaaacc taaactttaa caaatctgct cctgaattaa aaactggtat cagcgatgtt
     2041 tttgctaaaa atgatcttgc tgttgtggat gtcagaattg gtatgactag gaaatttggt
     2101 tatgtggatt ttgaatctgc tgaagacctg gagaaagcgt tggaactcac tggtttgaaa
     2161 gtctttggca atgaaattaa actagagaaa ccaaaaggaa aagacagtaa gaaagagcga
     2221 gatgcgagaa cacttttggc taaaaatctc ccttacaaag tcactcagga tgaattgaaa
     2281 gaagtgtttg aagatgctgc ggagatcaga ttagtcagca aggatgggaa aagtaaaggg
     2341 attgcttata ttgaatttaa gacagaagct gatgcagaga aaacctttga agaaaagcag
     2401 ggaacagaga tcgatgggcg atctatttcc ctgtactata ctggagagaa aggtcaaaat
     2461 caagactata gaggtggaaa gaatagcact tggagtggtg aatcaaaaac tctggtttta
     2521 agcaacctct cctacagtgc aacagaagaa actcttcagg aagtatttga gaaagcaact
     2581 tttatcaaag taccccagaa ccaaaatggc aaatctaaag ggtatgcatt tatagagttt
     2641 gcttcattcg aagacgctaa agaagcttta aattcctgta ataaaaggga aattgagggc
     2701 agagcaatca ggctggagtt gcaaggaccc aggggatcac ctaatgccag aagccagcca
     2761 tccaaaactc tgtttgtcaa aggcctgtct gaggatacca ctgaagagac attaaaggag
     2821 tcatttgacg gctccgttcg ggcaaggata gttactgacc gggaaactgg gtcctccaaa
     2881 gggtttggtt ttgtagactt caacagtgag gaggatgcca aagctgccaa ggaggccatg
     2941 gaagacggtg aaattgatgg aaataaagtt accttggact gggccaaacc taagggtgaa
     3001 ggtggcttcg ggggtcgtgg tggaggcaga ggcggctttg gaggacgagg tggtggtaga
     3061 ggaggccgag gaggatttgg tggcagaggc cggggaggct ttggagggcg aggaggcttc
     3121 cgaggaggca gaggaggagg aggtgaccac aagccacaag gaaagaagac gaagtttgaa
     3181 tagctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     3241 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3301 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3361 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3421 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3481 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3541 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3601 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3661 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3721 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3781 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3841 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3901 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3961 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4021 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4081 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4141 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4201 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4261 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4321 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4381 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4441 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4501 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4561 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4621 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4681 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4741 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4801 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4861 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4921 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4981 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5041 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5101 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5161 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5221 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5281 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5341 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5401 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5461 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5521 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5581 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5641 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5701 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5761 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5821 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5881 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5941 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6001 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6061 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6121 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6181 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6241 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6301 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6361 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     6421 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6481 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6541 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6601 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6661 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6721 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6781 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6841 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6901 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6961 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7021 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7081 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7141 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7201 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7261 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7321 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7381 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7441 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7501 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7561 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7621 tgacgtc
//
