LOCUS       V025923                 6107 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025923
VERSION     V025923
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"
     CDS             941..1597
                     /label="CDCA3(NM_001297603)"
                     /note="CDCA3(NM_001297603)"
                     /gene="CDCA3"
     CDS             1604..1669
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    1707..1931
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      1977..2405
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2419..2748
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             2815..3609
                     /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    3783..3904
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(3953..3969)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    3977..3993
                     /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(4001..4031)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4046..4067
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4355..4940)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5111..5971)
                     /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(5972..6076)
                     /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 aagcttggta ccgagctcgg atccgccacc atgggctcag ccaagagcgt
      961 cccagtcaca ccagcgcggc ctccgccgca caacaagcat ctggctcgag tggcggaccc
     1021 ccgttcacct agtgctggca tcctgcgcac tcccatccag gtggagagct ctccacagcc
     1081 aggcctacca gcaggggagc aactggaggg tcttaaacat gcccaggact cagatccccg
     1141 ctctcctact cttggtattg cacggacacc tatgaagacc agcagtggag accccccaag
     1201 cccactggtg aaacagctga gtgaagtatt tgaaactgaa gactctaaat caaatcttcc
     1261 cccagagcct gttctgcccc cagaggcacc tttatcttct gaattggact tgcctctggg
     1321 tacccagtta tctgttgagg aacagatgcc accttggaac cagactgagt tcccctccaa
     1381 acaggtgttt tccaaggagg aagcaagaca gcccacagaa acccctgtgg ccagccagag
     1441 ctccgacaag ccctcaaggg accctgagac tcccagatct tcagcctcct cagttccaga
     1501 ttttgaggaa gctgtcccag gaacctgtgg ccacagccat cccgtcagct gtgactccca
     1561 ggcagctcac cctgttatcg tggccagaga ggatgccctc gaggactaca aagaccatga
     1621 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc
     1681 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     1741 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     1801 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     1861 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     1921 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     1981 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2041 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2101 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2161 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2221 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2281 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2341 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2401 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2461 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2521 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2581 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2641 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     2701 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     2761 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     2821 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     2881 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     2941 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3001 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3061 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3121 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3181 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3241 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3301 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     3361 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3421 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3481 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     3541 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     3601 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     3661 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     3721 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     3781 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3841 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3901 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     3961 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4021 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4081 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4141 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4201 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4261 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4321 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4381 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4441 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4501 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4561 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4621 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4681 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4741 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4801 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4861 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     4921 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4981 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5041 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5101 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5161 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5221 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5281 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5341 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5401 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5461 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5521 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5581 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5641 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5701 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5761 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5821 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5881 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5941 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6001 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6061 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
