LOCUS       V025912                 8836 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025912
VERSION     V025912
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             964..4326
                     /label="WDR6(NM_018031)"
                     /note="WDR6(NM_018031)"
                     /gene="WDR6"
     CDS             4333..4398
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4436..4660
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4706..5134
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5148..5477
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5544..6338
                     /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    6512..6633
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6682..6698)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6706..6722
                     /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(6730..6760)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6775..6796
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7084..7669)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7840..8700)
                     /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(8701..8805)
                     /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 atccactagt ccagtgtggt ggaattcgcc
      961 accatggacg ctctcgagga ctacgtttgg ccgcgggcaa cctcggagct tatactcctc
     1021 ccagtgacgg gtctggagtg cgtgggggac cggctgttgg cgggtgaggg tcccgatgtc
     1081 ctggtgtaca gcttggactt tggtgggcat ctgcggatga taaagcgagt gcagaacctg
     1141 cttggccact atcttatcca tggcttccgg gtacggccag agcctaatgg agaccttgac
     1201 ttggaggcca tggtggctgt gtttggaagc aagggactcc gagttgtgaa aattagctgg
     1261 ggacagggcc acttctggga gctttggcgc tctggcctgt ggaacatgtc tgactggatt
     1321 tgggatgcac gctggcttga gggaaatata gccttggccc tgggccacaa ctcagtggtg
     1381 ctatatgacc ctgtagtagg gtgcatcctg caagaggtgc cctgcacaga caggtgcacc
     1441 ctctcttcag cctgcctgat tggagacgcc tggaaggagc tgaccatagt ggcaggtgct
     1501 gtttccaacc agctcttggt ctggtaccca gcaactgcct tagcagacaa caaacctgta
     1561 gcacctgacc gacgaatcag tgggcatgtg ggcatcatct tcagcatgtc atacctggaa
     1621 agcaagggat tgctggctac agcttcagaa gaccgaagcg ttcgtatctg gaaggtgggc
     1681 gacctgcgag tgcctggggg tcgggtgcag aatattgggc actgctttgg gcacagcgcc
     1741 cgtgtgtggc aggtcaagct tctagagaat taccttatca gtgcaggaga ggattgtgtc
     1801 tgcttggtgt ggagccatga aggtgagatc ctccaggcct ttcggggaca ccagggacgt
     1861 gggatccggg ccatagctgc ccatgagagg caggcctggg tgatcactgg gggtgatgac
     1921 tcaggcattc ggctgtggca cttggtaggg cgtgggtacc ggggattggg ggtctcggct
     1981 ctctgcttca agtcccgtag taggccaggt acactcaagg ctgtgactct ggctggctct
     2041 tggcgactgc tggcagtgac tgatacaggg gccctgtatc tctatgacgt cgaggtcaag
     2101 tgctgggagc agctgctaga ggataaacat ttccagtcct actgcctgct ggaggcagct
     2161 cctggtcccg agggcttcgg attgtgtgct atggccaatg gggaaggtcg tgtcaaggtt
     2221 gtccccatca acactccaac tgctgctgtg gaccagaccc tgtttcctgg gaaggtgcac
     2281 agcttgagct gggccctgcg tggttatgag gagctcctgt tgctggcatc gggccctggc
     2341 ggggtagtag cttgcctaga gatctcagcc gcaccctctg gcaaggccat ctttgtcaag
     2401 gaacgttgtc ggtacctgct gcccccaagc aagcagagat ggcacacatg cagtgccttc
     2461 ctacccccag gtgacttcct ggtgtgtggt gaccgccggg gctctgtgct gctattcccc
     2521 tccagaccag gtctgctcaa ggaccctggg gtgggaggca aggctcgggc tggtgctggg
     2581 gcacctgtag tgggtagtgg tagtagtggg ggtgggaatg ctttcactgg gttgggccca
     2641 gtgtctaccc tgccctctct gcacgggaag cagggtgtga cctcagtcac atgccatggt
     2701 ggctatgtgt ataccacagg gcgtgatgga gcctactacc agctgtttgt acgagacggc
     2761 cagctccagc cagtcctaag gcagaagtcc tgtcgaggca tgaactggct agctgggctc
     2821 cgtatagtgc ccgatgggag catggttatc ctgggtttcc atgccaatga gtttgtggtg
     2881 tggaaccctc ggtcacacga gaagctgcac atcgtcaact gtggtggagg gcaccgttcg
     2941 tgggcattct ctgatactga ggcggccatg gcctttgctt acctcaagga tggggatgtc
     3001 atgctgtaca gggctctggg tggctgcacc cggccacacg tgattctccg ggagggtctg
     3061 catggccgtg agatcacttg tgtaaagcgt gtgggcacca ttaccctggg gcctgaatat
     3121 ggagtgccca gcttcatgca gcctgatgac ctggagcctg gcagtgaggg gcccgacttg
     3181 actgacattg tgatcacatg tagtgaggac actactgtct gtgtcctagc actccctaca
     3241 accacaggct cagcccacgc actcacagct gtttgtaacc atatctcctc ggtacgtgct
     3301 gtggctgtgt ggggcattgg caccccaggt ggccctcagg atcctcagcc aggcctgact
     3361 gcccatgtgg tgtctgcggg ggggcgggct gagatgcact gcttcagcat catggttact
     3421 ccggacccca gcaccccaag ccgcctcgcc tgccatgtca tgcacctttc gtcccaccgg
     3481 ctagatgagt attgggaccg gcaacgcaat cggcatcgga tggttaaggt agacccagag
     3541 accaggtaca tgtcccttgc tgtgtgtgaa cttgaccagc ccggccttgg cccccttgtg
     3601 gctgcagcct gtagtgatgg ggccgtaagg ctctttcttt tgcaggattc tgggcggatt
     3661 ctgcagctcc ttgctgaaac cttccaccat aagcgatgtg tcctcaaggt ccactccttt
     3721 acacacgagg cacccaacca gaggcggagg ctcctcctgt gcagcgcagc tactgatggc
     3781 agcctggctt tctgggatct caccaccatg ctagaccatg actccactgt cctggagcct
     3841 ccagtggatc ctgggcttcc ctaccggctt ggcaccccct ccctgactct ccaggcccac
     3901 agctgtggta tcaacagcct gcacaccttg cccacccgtg agggccacca tctcgtggcc
     3961 agtggcagtg aagatggatc cctccatgtc ttcgtgcttg ctgtggagat gctacagcta
     4021 gaagaggctg tgggagaggc tgggctggta ccccagctgc gtgtgctaga ggaatactct
     4081 gtcccctgtg cacatgctgc ccatgtgaca ggcctcaaga tcctaagccc aagcatcatg
     4141 gtctcagcct ccattgatca acggctgacc ttctggcgtc tggggcatgg tgaacccacc
     4201 ttcatgaata gcactgtgtt ccatgtgcct gatgtggctg acatggactg ctggcctgtg
     4261 agccctgagt ttggccaccg ttgtgccctt gggggtcagg ggcttgaggt ttacaactgg
     4321 tatgacctcg aggactacaa agaccatgac ggtgattata aagatcatga catcgactac
     4381 aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc ctcgactgtg
     4441 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     4501 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     4561 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     4621 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     4681 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4741 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4801 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4861 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4921 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4981 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     5041 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     5101 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     5161 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5221 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5281 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     5341 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5401 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5461 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     5521 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     5581 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     5641 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5701 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     5761 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5821 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5881 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5941 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     6001 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     6061 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     6121 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     6181 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     6241 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     6301 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     6361 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     6421 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     6481 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     6541 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6601 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     6661 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6721 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6781 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6841 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6901 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6961 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     7021 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     7081 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     7141 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     7201 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     7261 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7321 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7381 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7441 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7501 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     7561 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7621 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7681 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7741 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7801 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7861 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7921 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7981 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     8041 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8101 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     8161 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8221 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8281 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8341 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8401 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8461 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8521 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8581 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8641 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8701 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8761 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8821 agtgccacct gacgtc
//
