LOCUS       V045462                 7683 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045462
VERSION     V045462
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2363
                     /label="Zfp454(NM_172794)"
                     /note="Zfp454(NM_172794)"
                     /gene="Zfp454"
     CDS             2370..2435
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2459..3047
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3113..3161
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3363..3791
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3805..4134
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4201..4995
                     /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    5171..5292
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5341..5357)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5365..5381
                     /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(5389..5419)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5434..5455
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5743..6331)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6502..7362)
                     /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(7363..7467)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc tgtcagccag ctcccggccc
      781 tggtccagga cttggtgacc ttcaaggatg tggccgtgct tttcacccag gaggagtggg
      841 gtcagctgag ctctgcccag agggccttgt atcaggatgt gatgctagag aactacagta
      901 acctggtctc actggcaggt ctcttaggat cccaaccaga tatgttcttt cccctggagg
      961 aagtggaaga atgtgtgtca gaggaggctc ctgagggctt tgttctggat gccgcagatg
     1021 aacctcactt gggtcctcga tctgcgtgtg aagctcaggt tccggttaca ggggaaatta
     1081 cagtggccat gagtaagaaa ttcctcaagg cagatatccc tgagaaagag tcggatcaat
     1141 ggatagcaaa ggaaagagca tgtgctaact tgttcaattg gcagtgccag gaaggtcagg
     1201 aggtgaagtt gcagcaagtg gtcctcaccc agcagaacac tccatcaaaa ctgagtgagc
     1261 agagagtcag agcagagagc tcatcaccca tccaaagtca gagatctcaa gcaagcaaaa
     1321 cggcttttga atgcagcgaa tgcgggaaag ccttctctaa gagttccacc cttaagaagc
     1381 atcagaaact tcataccgaa aaactcaacc caagtcagaa atcaccaatg aaagagaagc
     1441 gatacaagtg ccgagaatgt gggaaagcct tccaccagag tacacacctc atccatcacc
     1501 aaagagtcca cactggcgag aaaccatacc agtgcaagga ctgtggcaag gccttctcag
     1561 tgagctcttc cctttcctac caccagaaaa tccataccgg agaaaagccc tttgaatgta
     1621 atgtttgtgg aaaggccttt atccggaaca tacatctttc ccaccatcat agaatgcaca
     1681 ctggagagaa acctttccaa tgcaaccttt gtgacaaagc tttcgtttgc agagcacatc
     1741 ttaccaaaca ccagcacatt cacagtggca agaagcccta taaatgtaat gagtgtggga
     1801 aagccttcaa tcagagtaca agttttctgc agcatcagag gatccacact ggagagaaac
     1861 cctttgaatg caacgagtgt ggcaaggcct tcagggtgaa ctcttccctt actgaacacc
     1921 agagaatcca cactggggag aaaccatatc aatgtatcga atgtggcaaa gctttcaggg
     1981 ataactcatc ctttgcacgc catcggaaaa ttcacactgg agagaaacct tacagatgtg
     2041 gcttgtgtga gaaggctttc agggaccagt cagccctagc acagcatcag agaactcaca
     2101 ctggggaaaa gccttacacg tgtaacatat gtgagaaagc cttcagtgac cattcggccc
     2161 ttacccaaca taagagaatt cacacgagag aaaaaccgta caaatgtaaa acctgtggga
     2221 aagcttttat ccgaagcaca cacctcattc aacaccagag gattcacacg ggagagaaac
     2281 cttacaaatg taacacatgc gggaaagcct tcaaccagac tgcaaacctc gctcagcacc
     2341 agaggcatca cactggagga aagggatccg actacaaaga ccatgacggt gattataaag
     2401 atcatgacat cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa
     2461 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     2521 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     2581 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     2641 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     2701 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     2761 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2821 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     2881 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     2941 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     3001 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg
     3061 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     3121 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     3181 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     3241 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     3301 cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc
     3361 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3421 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3481 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     3541 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3601 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3661 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3721 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3781 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3841 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3901 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3961 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4021 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     4081 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4141 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     4201 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4261 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4321 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4381 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4441 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4501 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4561 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4621 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4681 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4741 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4801 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4861 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4921 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4981 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa
     5041 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5101 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5161 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5221 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5281 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5341 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5401 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5461 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5521 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5581 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5641 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5701 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5761 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5821 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5881 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     5941 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6001 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6061 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6121 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6181 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6241 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6301 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6361 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6421 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6481 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6541 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6601 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6661 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6721 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6781 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6841 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6901 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6961 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7021 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7081 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7141 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7201 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7261 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7321 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7381 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7441 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     7501 cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc
     7561 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     7621 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     7681 agg
//
