LOCUS       V023951                 6608 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023951
VERSION     V023951
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..2098
                     /label="TFDP2(NM_001375776)"
                     /note="TFDP2(NM_001375776)"
                     /gene="TFDP2"
     CDS             2105..2170
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2208..2432
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2478..2906
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2920..3249
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3316..4110
                     /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    4284..4405
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4454..4470)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4478..4494
                     /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(4502..4532)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4547..4568
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4856..5441)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5612..6472)
                     /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(6473..6577)
                     /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 atgattataa gcacaccaca
      961 gagactaacc agttcaggaa gtgttctgat tgggagtcca tatacccctg caccagcaat
     1021 ggttactcag acacacatag cagaagctac tggctgggtc cctggtgata gaaaacgggc
     1081 tagaaaattt atagactctg atttttcaga aagtaaacga agcaaaaaag gagataaaaa
     1141 tgggaaaggc ttgagacact tttcaatgaa agtgtgtgag aaagttcaac gaaaaggtac
     1201 aacatcgtac aatgaagtcg ctgatgagct ggtgtcagag ttcaccaatt caaataacca
     1261 tttggctgct gattcgcagg cttatgatca gaagaacatt aggcgaagag tttatgatgc
     1321 tttaaatgtg ctaatggcaa tgaacataat ttcaaaggaa aaaaaagaaa tcaagtggat
     1381 tggcctgcct accaattctg ctcaggaatg tcagaatctg gagatagaga agcagaggcg
     1441 gatagaacgg ataaagcaga agcgggccca gctgcaagaa cttctcctac agcaaatcgc
     1501 tttcaaaaac ctggtacaga gaaatcgaca aaatgagcag caaaaccagg gcccgccggc
     1561 tctgaactct accattcagc tgccattcat aatcatcaat acaagcagaa aaacagtcat
     1621 agattgcagc atctccagtg acaagtttga gtatcttttc aattttgaca acacctttga
     1681 gatccatgat gacatagaag tactaaagcg gatgggaatg tcgtttggcc tggagtcagg
     1741 caaatgctct ctggaggatc tgaaacttgc gaaatccctg gtgccaaagg ctttagaagg
     1801 ttatatcaca gatatctcca caggaccttc ttggttaaat cagggactac ttctgaactc
     1861 tacccaatca gtttcaaatt tagacctgac cactggtgcc accttacccc agtcaagtgt
     1921 aaaccaaggg ttatgcttgg atgcagaagt ggccttagca actgggcagt tcctggcccc
     1981 aaacagtcac cagtccagca gtgcggcctc tcactgctcc gagtcccgag gcgagacccc
     2041 ctgttcgttc aatgatgaag atgaggaaga tgatgaggag gattcctcct ccccagaact
     2101 cgaggactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     2161 cgatgacaag tagtgagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     2221 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     2281 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2341 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     2401 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     2461 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2521 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2581 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2641 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2701 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2761 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     2821 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     2881 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     2941 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3001 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3061 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3121 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3181 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3241 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     3301 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3361 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3421 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3481 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3541 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3601 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3661 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3721 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3781 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3841 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3901 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3961 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4021 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4081 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4141 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4201 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4261 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     4321 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4381 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     4441 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4501 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4561 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4621 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4681 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4741 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4801 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4861 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4921 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4981 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5041 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5101 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5161 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5221 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5281 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5341 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg
     5401 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5461 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5521 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5581 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5641 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5701 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5761 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5821 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5881 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5941 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6001 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6061 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6121 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6181 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6241 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6301 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6361 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6421 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6481 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6541 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6601 ctgacgtc
//
