LOCUS       V028760                 7689 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028760
VERSION     V028760
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..2369
                     /label="EYA2(NM_005244)"
                     /note="EYA2(NM_005244)"
                     /gene="EYA2"
     CDS             2376..2441
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2465..3053
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3119..3167
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3369..3797
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3811..4140
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4207..5001
                     /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    5177..5298
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5347..5363)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5371..5387
                     /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(5395..5425)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5440..5461
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5749..6337)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6508..7368)
                     /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(7369..7473)
                     /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 ccaccatggt agaactagtg atctcaccca
      781 gcctcactgt aaacagcgat tgtctggata aactgaagtt taaccgtgct gacgctgctg
      841 tgtggactct gagtgacaga caaggcatca ccaaatcggc ccccctgaga gtgtcccagc
      901 tcttctccag atcttgccca cgtgtcctcc cccgccagcc ttccacagcc atggcagcct
      961 acggccagac gcagtacagt gcggggatcc agcaggctac cccctataca gcttacccac
     1021 ctccagcaca agcctatgga atcccttcct acagcatcaa gacagaagac agcttgaacc
     1081 attcccctgg ccagagtgga ttcctcagct atggctccag cttcagcacc tcacccactg
     1141 gacagagccc atacacctac cagatgcacg gcacaacagg gttctatcaa ggaggaaatg
     1201 gactgggcaa cgcagccggt ttcgggagtg tgcaccagga ctatccttcc taccccggct
     1261 tcccccagag ccagtacccc cagtattacg gctcatccta caaccctccc tacgtcccgg
     1321 ccagcagcat ctgcccttcg cccctctcca cgtccaccta cgtcctccag gaggcatctc
     1381 acaacgtccc caaccagagt tccgagtcac ttgctggtga atacaacaca cacaatggac
     1441 cttccacacc agcgaaagag ggagacacag acaggccgca ccgggcctcc gacgggaagc
     1501 tccgaggccg gtctaagagg agcagtgacc cgtccccggc aggggacaat gagattgagc
     1561 gtgtgttcgt gtgggacttg gatgagacaa taattatttt tcactcctta ctcacgggga
     1621 catttgcatc cagatacggg aaggacacca cgacgtccgt gcgcattggc cttatgatgg
     1681 aagagatgat cttcaacctt gcagatacac atctgttctt caatgacctg gaggattgtg
     1741 accagatcca cgttgatgac gtctcatcag atgacaatgg ccaagattta agcacataca
     1801 acttctccgc tgacggcttc cacagttcgg ccccaggagc caacctgtgc ctgggctctg
     1861 gcgtgcacgg cggcgtggac tggatgagga agctggcctt ccgctaccgg cgggtgaagg
     1921 agatgtacaa tacctacaag aacaacgttg gtgggttgat aggcactccc aaaagggaga
     1981 cctggctaca gctccgagct gagctggaag ctctcacaga cctctggctg acccactccc
     2041 tgaaggcact aaacctcatc aactcccggc ccaactgtgt caatgtgctg gtcaccacca
     2101 ctcaactaat tcctgccctg gccaaagtcc tgctatatgg cctggggtct gtgtttccta
     2161 ttgagaacat ctacagtgca accaagacag ggaaggagag ctgcttcgag aggataatgc
     2221 agagattcgg cagaaaagct gtctacgtgg tgatcggtga tggtgtggaa gaggagcaag
     2281 gagcgaaaaa gcacaacatg cctttctggc ggatatcctg ccacgcagac ctggaggcac
     2341 tgaggcacgc cctggagctg gagtatttag gatccgacta caaagaccat gacggtgatt
     2401 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct
     2461 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     2521 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     2581 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     2641 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     2701 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     2761 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     2821 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     2881 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     2941 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     3001 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     3061 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     3121 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     3181 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     3241 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     3301 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     3361 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3421 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3481 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     3541 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3601 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3661 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3721 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     3781 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3841 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3901 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3961 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4021 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4081 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4141 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4201 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4261 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4321 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4381 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4441 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4501 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4561 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4621 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4681 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4741 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4801 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4861 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4921 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4981 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     5041 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5101 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5161 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5221 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5281 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5341 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5401 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5461 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5521 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5581 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5641 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5701 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5761 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5821 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5881 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5941 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6001 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6061 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6121 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6181 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6241 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6301 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6361 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6421 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6481 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6541 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6601 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6661 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6721 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6781 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6841 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6901 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6961 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7021 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7081 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7141 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7201 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7261 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7321 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7381 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7441 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7501 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     7561 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     7621 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     7681 ctgcttagg
//
