LOCUS       V045333                 7432 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045333
VERSION     V045333
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             932..2911
                     /label="FOXK2(NM_004514)"
                     /note="FOXK2(NM_004514)"
                     /gene="FOXK2"
     CDS             2933..2998
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3027..3251
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3297..3725
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3739..4068
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4135..4929
                     /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    5105..5226
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5275..5291)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5299..5315
                     /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(5323..5353)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5368..5389
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5677..6265)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6436..7296)
                     /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(7297..7401)
                     /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 gctggctagt
      901 taagcttggt accgagctcg gatccgccac catggcagct gccgctgcag cattgtctgg
      961 agctggaact cctccagctg gaggtggtgc tggtggtgga ggagcaggag gaggaggatc
     1021 tcctccagga ggatgggcag tggcaagact cgagggccgc gagttcgagt atctgatgaa
     1081 gaagcgctcg gtgaccatcg gccgcaactc gtcgcagggc tcggtggacg tgagcatggg
     1141 ccactcgagc ttcatctccc ggcgccacct cgagatcttc acgcccccgg gcggcggcgg
     1201 ccatggcggg gccgctccgg agctgccgcc cgcgcagccc aggcccgacg ccggcggcga
     1261 cttctacctg cgctgcttgg gcaagaacgg ggtattcgtg gacggcgtgt tccagaggcg
     1321 cggggcgccg ccgctgcagc tgccgcgcgt gtgcacattc aggttcccga gcacaaacat
     1381 caagataacg ttcactgccc tgtccagcga gaagagagag aagcaggagg cgtctgagtc
     1441 tccagtgaag gccgtacagc cacacatctc gcccctgacc atcaacattc cagacaccat
     1501 ggcccacctc atcagccctc tgccctcccc cacgggaacc atcagcgctg caaactcctg
     1561 cccctccagc ccccggggag cggggtcttc agggtacaag gtgggccgag tgatgccatc
     1621 tgacctcaat ttaatggctg acaactcaca gcctgaaaat gaaaaggaag cttcaggtgg
     1681 agacagcccg aaggatgatt caaagccgcc ttactcctac gcgcagctga tagttcaggc
     1741 gattacgatg gctcccgaca aacagctcac cctgaacggg atttatacac acatcactaa
     1801 aaattatccc tactacagga ctgcggacaa gggctggcag aattcaattc gccacaatct
     1861 ctctctgaat cgttatttca tcaaagtgcc gcgttcccag gaagaaccag gcaaaggctc
     1921 gttctggagg atagacccag cctctgaaag caaattaata gaacaggctt ttaggaaacg
     1981 acggcctagg ggcgtgccct gctttagaac ccctctggga ccgctctctt ctaggagtgc
     2041 cccagcctct cccaatcacg cgggagtgct gtctgctcac tctagtggcg cccagacccc
     2101 tgagagcctg tcgagggaag gttcgccggc ccccctggag cctgagcctg gcgctgcaca
     2161 gcccaaactc gctgtcatcc aggaagcccg gtttgcccag agcgccccag ggtcacctct
     2221 gtccagtcag ccagtcttaa tcaccgtcca gcggcagcta ccacaggcca tcaagcctgt
     2281 cacctacact gtggccaccc cagtgaccac ctcgacctcc cagccacccg tcgtgcagac
     2341 ggttcacgtc gtccaccaga tcccagcggt gtcggtcacc agtgtggccg gactggcccc
     2401 agcgaacacg tacactgtct ctggacaagc tgtggtcacc ccggcagccg tgctggcccc
     2461 tcctaaggca gaggcccagg agaatggaga ccacagggaa gtcaaagtga aagtagagcc
     2521 tattcccgcc attggccacg ccacgctcgg cactgccagc cggatcattc agacggcaca
     2581 gaccaccccg gtccagacgg tgaccatagt acaacaggca cctctaggtc aacaccagct
     2641 accaataaaa actgtaacac aaaacggcac tcacgtggca tcagtcccca ctgcggtcca
     2701 cggccaggtg aacaatgccg cggcgagtcc tttgcacatg ttggcaacac acgcatccgc
     2761 atcggcctcc ctgcccacaa agcgccacaa cggtgaccag ccggagcagc cggagctgaa
     2821 gcggatcaag acagaagacg gcgagggcat cgtcattgcc ctgagcgtgg acacgccacc
     2881 ggcagccgta agggaaaagg gtgtccagaa cctcgagtct agagggccct tcgactacaa
     2941 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagtg
     3001 agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3061 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3121 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3181 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3241 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3301 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3361 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3421 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3481 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3541 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3601 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3661 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3721 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3781 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3841 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3901 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3961 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4021 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4081 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4141 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4201 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4261 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4321 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4381 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4441 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4501 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4561 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4621 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4681 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4741 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4801 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4861 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4921 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     4981 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5041 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5101 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5161 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5221 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     5281 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5341 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5401 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5461 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5521 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5581 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5641 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5701 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5761 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5821 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5881 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5941 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6001 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6061 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     6121 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6181 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6241 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6301 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6361 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6421 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6481 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6541 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6601 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6661 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6721 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6781 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6841 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6901 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6961 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7021 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7081 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7141 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7201 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7261 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7321 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7381 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
