LOCUS       V037405                10186 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037405
VERSION     V037405
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..5706
                     /label="HECW2(NM_020760)"
                     /note="HECW2(NM_020760)"
                     /gene="HECW2"
     polyA_signal    5786..6010
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      6056..6484
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6498..6827
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6894..7688
                     /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    7862..7983
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8032..8048)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8056..8072
                     /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(8080..8110)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8125..8146
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8434..9019)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9190..10050)
                     /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(10051..10155)
                     /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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atggctagtt cagcccggga gcacctgctt
     1021 tttgtgaggc gtcgaaatcc ccagatgcgg tacacattga gcccagagaa cctccagagc
     1081 cttgccgccc agagctccat gccagagaac atgaccctgc agcgggccaa cagcgacacc
     1141 gacctggtga cttctgagag ccgctccagc ttaactgcca gcatgtacga gtacacgctg
     1201 gggcaagccc agaacctcat tatcttctgg gacattaaag aggaggtgga ccccagtgat
     1261 tggattggac tttatcatat agatgagaat tctccagcca acttctggga ttctaagaac
     1321 aggggtgtga ctggaacaca aaaagggcaa attgtatgga gaattgagcc tgggccctat
     1381 ttcatggaac cggagataaa aatctgtttt aaatattacc acggcattag tggagccctg
     1441 cgagccacga ccccctgcat caccgtgaag aacccagctg tgatgatggg ggcagaaggc
     1501 atggagggag gtgcttcagg aaacctgcat tctcgaaaac ttgttagctt tacattgtca
     1561 gatcttaggg cagttgggct aaagaaaggg atgttcttca atcctgaccc ttatcttaag
     1621 atgtcaattc agccaggaaa gaagagcagt ttccccacct gtgcccacca cgggcaggag
     1681 agacggtcta ctatcatcag taacaccacc aatccaattt ggcaccgaga gaaatattcc
     1741 ttttttgcac ttcttactga tgtcttagaa attgaaatta aagacaaatt tgccaagagc
     1801 cgtcccatca tcaagcgttt tctggggaaa ctaaccattc cagtccagag gctgctggag
     1861 cgacaagcca tcggtgatca aatgctcagc tacaaccttg gcagaaggct cccagctgac
     1921 cacgtgagtg ggtacctcca gtttaaagtg gaggttacgt cttctgttca tgaagatgcc
     1981 tctccagaag ctgttggcac aatacttgga gtcaattctg tgaatggaga cttaggtagc
     2041 ccttccgatg acgaggacat gccagggagc catcacgaca gccaggtgtg ctctaatggg
     2101 ccagtttctg aggacagtgc tgccgatgga acccccaagc attcattcag gactagctcc
     2161 actctggaaa tagacacaga ggaattaacc tctacgtctt caaggacctc acctcccaga
     2221 ggacgtcagg attcactcaa tgattactta gatgctatcg aacacaatgg ccactccagg
     2281 ccggggacag cgacctgctc tgagaggtcc atgggtgcct ctccgaaact gaggagtagt
     2341 tttcccactg acacaagact caatgccatg cttcatattg attcagatga agaagatcac
     2401 gagttccagc aagacctggg ttacccatct tccttggagg aggagggagg cctgatcatg
     2461 tttagcaggg catccagagc tgatgatgga agcctgacat ctcagacaaa gctggaggac
     2521 aaccctgttg agaatgagga agcctccaca cacgaagctg cttcctttga ggataagcca
     2581 gaaaatcttc cagagcttgc agagagctcc ttacctgcag gcccagcccc agaggaaggt
     2641 gaaggcggcc cagagcctca acccagtgct gaccagggca gtgctgaact gtgtggctct
     2701 caagaggtag atcagcccac aagtggcgca gacacaggga cttccgatgc atcaggagga
     2761 agccgaagag ccgtcagtga gaccgagtct ctcgatcagg gctctgagcc ttcccaggtg
     2821 tcctctgaaa cagaacccag tgatcctgcc aggacagaga gtgtgagcga agccagcacc
     2881 aggcctgagg gagagagtga cctggaatgc gctgacagct cctgcaatga gagtgtgacc
     2941 acgcagctgt cctctgtgga cacacggtgc tcctctcttg agagcgcacg gtttcctgaa
     3001 accccagcct tttcttctca ggaggaggaa gacggagcct gtgcagcaga gcccaccagc
     3061 agtggccctg ccgaagggtc gcaggaatcc gtgtgcactg ctggttcttt acctgtggta
     3121 caggtgccca gtggggagga tgaagggcca ggggcagaat cggccactgt acctgaccag
     3181 gaggagctgg gggaggtctg gcagcggagg gggagcctgg agggagctgc agctgctgcc
     3241 gagagcccac cgcaagaaga aggcagtgct ggggaggccc aaggcacctg tgaaggggca
     3301 actgcccagg aggagggcgc tactggaggt tctcaagcca acggccacca gccactgcga
     3361 tcactacctt cagtgcgcca ggatgttagc cggtaccaga gggtggacga ggctctccca
     3421 ccaaactggg aggcacgcat tgacagccac ggcaggatct tctacgtgga tcacgtaaac
     3481 agaaccacga cgtggcagcg accgacagct cccccagccc cgcaggtgct gcagagatct
     3541 aactccatac agcaaatgga gcagctgaac cggcgatatc agagtatccg cagaaccatg
     3601 accaatgaga ggcctgagga aaataccaat gctattgatg gggcagggga ggaagctgac
     3661 tttcaccaag ccagtgcaga tttccgacgg gagaacatcc tgcctcactc tacctccaga
     3721 tccaggatca cgttgctgtt acagtctccc cccgtgaagt tcctcatcag cccagagttc
     3781 ttcaccgtgc tgcattctaa ccctagtgcc taccgcatgt ttacaaacaa cacgtgtttg
     3841 aagcacatga tcaccaaagt ccggagggac acccaccact ttgaacgcta ccagcataac
     3901 cgcgaccttg tgggattcct caacatgttc gcgaacaaac agctagagct gccgcgggga
     3961 tgggaaatga aacatgatca ccagggcaag gcattctttg tggaccacaa ctcccgcacc
     4021 accactttca ttgatccccg gctcccactt cagagcagta gacccacaag tgcgctggtt
     4081 catcggcaac acctgacaag gcaacgcagc cacagtgcgg gtgaggtagg agaagattct
     4141 cgacatgcag gaccaccagt tcttcccagg ccatccagta cattcaatac agtcagtagg
     4201 ccacagtacc aggacatggt tccagtggct tacaatgaca agattgttgc atttctacgt
     4261 caacccaata tctttgaaat tctacaggag cgtcaacctg atcttaccag gaatcactca
     4321 ctcagggaga agatccaatt tatccgaact gaagggactc caggattggt gcgcctttca
     4381 agcgatgcag accttgttat gttgctgagc ttatttgaag aagagataat gtcgtatgtg
     4441 cctccacatg ccttactcca ccccagctac tgtcagtccc cacgtggctc tcccgtgtca
     4501 tctcctcaga actcgccagg tacccagcgt gccaatgccc gggctccagc cccttacaag
     4561 cgggatttcg aagccaaact gaggaacttt tacaggaagt tagagactaa aggatatgga
     4621 caaggcccag ggaagttaaa gttaattatc cgaagagatc acttactaga agatgctttt
     4681 aatcagatta tgggctactc cagaaaagac ctgcagagaa ataagctata tgtcaccttc
     4741 gttggggagg aagggctgga ttacagtggg ccttctagag agtttttctt cctggtatcc
     4801 agagaactct ttaacccata ttatggctta tttgaatatt cagccaatga cacatacaca
     4861 gtacaaataa gtcctatgtc tgcttttgta gacaatcacc atgaatggtt ccgattcagt
     4921 ggtaggatcc ttggtcttgc actaatacac cagtatttgt tggatgcctt cttcacacgg
     4981 cccttttata aggctcttct cagaattcta tgtgacctga gtgacctaga ataccttgat
     5041 gaagagttcc atcagagcct gcagtggatg aaagacaatg atatccatga catcctagac
     5101 ctcacgttca ctgtgaacga agaagtattt gggcagataa ctgaacgaga attaaagcca
     5161 gggggtgcca atatcccagt tacagagaag aacaagaagg agtacatcga gaggatggtg
     5221 aagtggagga ttgagagggg tgttgtacag caaacagaga gcttagtgcg tggcttctat
     5281 gaggtggtgg atgccaggct ggtatctgtt tttgatgcaa gagaactgga attggtcatc
     5341 gcaggcacag ctgaaataga cctaagtgat tggagaaaca acacagaata tagaggagga
     5401 taccatgaca atcatattgt aattcggtgg ttctgggctg cagtggaaag attcaacaat
     5461 gaacaacgac taaggttgtt acagtttgtt acaggcacat ccagcattcc ctatgaagga
     5521 tttgcttcac tccgagggag taacggccca agaagattct gtgtggagaa atgggggaaa
     5581 atcactgctc ttcccagagc gcatacatgt tttaaccgtc tggatctgcc tccctaccca
     5641 tccttttcca tgctttatga aaaactgttg acagcagttg aagaaaccag tacttttgga
     5701 cttgagtgag aattctgcag atatccagca cagtggcggc cgctcgagtc tagagggccc
     5761 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     5821 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     5881 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     5941 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     6001 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     6061 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     6121 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     6181 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     6241 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     6301 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     6361 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     6421 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6481 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     6541 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     6601 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     6661 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     6721 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     6781 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     6841 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     6901 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     6961 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     7021 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     7081 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     7141 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     7201 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     7261 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     7321 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     7381 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     7441 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     7501 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     7561 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     7621 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     7681 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     7741 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     7801 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     7861 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     7921 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     7981 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     8041 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     8101 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     8161 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     8221 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     8281 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     8341 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     8401 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     8461 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     8521 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     8581 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     8641 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     8701 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     8761 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     8821 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     8881 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     8941 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     9001 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     9061 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     9121 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     9181 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     9241 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     9301 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     9361 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     9421 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     9481 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     9541 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     9601 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     9661 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     9721 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     9781 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     9841 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     9901 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     9961 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
    10021 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
    10081 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
    10141 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
