LOCUS       V030444                 7875 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030444
VERSION     V030444
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     166..182
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        343..722
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        723..926
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        952..970
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             1029..4022
                     /label="Epha7(NM_010141)"
                     /note="Epha7(NM_010141)"
                     /gene="Epha7"
     CDS             4041..4070
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             4089..4112
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(4132..4148)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    4159..4781
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(4930..5518)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    5847..5894
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(6126..6920)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     promoter        complement(6955..7312)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(7314..7418)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      7445..7873
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg
     1021 cgatcgccat ggttgttcaa actcggttcc cttcgtggat tattttgtgt tacatctggc
     1081 tgcttggctt tgcacacacg ggggaggcgc aggctgcgaa ggaagtacta ttactggact
     1141 cgaaagcaca acaaacagaa ttggaatgga tttcctctcc acccagtggg tgggaagaaa
     1201 ttagtggttt ggatgagaac tacactccga taagaacata ccaggtgtgc caggtcatgg
     1261 agcccaacca gaacaactgg ctgcggacta actggatttc taaaggcaac gcacaaagga
     1321 tttttgtaga attgaaattc accttgaggg attgtaatag tcttcccgga gtcctgggaa
     1381 cttgcaagga aacgtttaat ttgtactatt atgaaacaga ctacgacacc ggcaggaata
     1441 tacgagaaaa cctttatgtt aaaatagaca ccattgctgc agatgaaagt ttcacacaag
     1501 gtgaccttgg tgaaagaaag atgaagctga acactgaggt gagagagatt ggacctttgt
     1561 ccaaaaaggg attctatctt gcctttcagg atgtaggggc ttgcatagca ttggtttctg
     1621 tcaaagtgta ctacaagaag tgctggtcca ttgttgagaa cttagctgtc tttccagata
     1681 cagtgactgg ttcggaattt tcctccttag tcgaggtccg tgggacatgt gtcagcagtg
     1741 ccgaggaaga ggcagaaaat tcccccagaa tgcattgcag tgcagaagga gagtggctag
     1801 tacccattgg aaaatgcatc tgcaaagcag gctatcagca aaaaggggac acttgcgaac
     1861 cctgtggccg caggttctac aaatcttcct ctcaggatct ccagtgttct cgttgtccaa
     1921 cccacagctt ctctgaccga gaaggatcat ccaggtgtga atgtgaagat gggtactaca
     1981 gagctccttc tgatccacca tacgttgcat gcacgaggcc tccctctgca ccacagaacc
     2041 ttattttcaa tatcaatcaa acgactgtaa gtttggaatg gagtcctccg gctgacaacg
     2101 ggggaagaaa cgatgtcacc tacagaatac tgtgtaagcg gtgcagttgg gaacagggag
     2161 aatgtgtgcc atgcggaagt aacattggat acatgcccca gcagacggga ttagaggata
     2221 actatgtcac tgtcatggac ctacttgccc atgcaaatta cactttcgaa gttgaagctg
     2281 taaatggagt ttcggactta agcagatccc agaggctctt cgctgctgtt agcatcacca
     2341 ccggtcaagc agctccctcg caagtgagtg gagtcatgaa ggagcgagta ctgcagcgga
     2401 gtgtgcagct ttcctggcag gagccggagc atcccaatgg agtcatcacg gaatatgaaa
     2461 tcaagtatta tgagaaagat caacgggaaa ggacgtactc aacactcaaa accaagtcca
     2521 cctccgcctc cattaataat ctgaaaccgg gaacagtgta cgtctttcag atccgggcgg
     2581 tcactgctgc cggttatgga aactacagcc ctaggcttga tgttgccaca cttgaggaag
     2641 cttcaggtaa aatgtttgaa gcgacagcag tctccagtga acagaatcct gtcatcataa
     2701 ttgctgtagt ggctgtagca gggaccatca tcttggtgtt catggtgttc ggcttcatca
     2761 ttggaagaag gcactgtggt tatagcaagg ctgaccaaga aggggatgaa gaactctact
     2821 ttcattttaa atttccaggc accaaaacct acattgaccc tgaaacctat gaggacccaa
     2881 atagagctgt ccatcaattc gccaaggagc tagatgcctc ctgtattaaa attgagcgtg
     2941 tgattggtgc aggagaattt ggagaagttt gcagtggtcg tttgaaactt ccggggaaga
     3001 gagatgttgc agtggccata aaaaccctga aagttggtta cacagaaaag caaaggaggg
     3061 actttttatg cgaagcaagc atcatggggc aatttgacca cccaaatgtc gtccatttgg
     3121 aaggggttgt tacaagaggg aagcctgtca tgattgtgat agagttcatg gagaatggag
     3181 ccctggatgc atttctcagg aaacacgatg ggcagtttac agtcattcag ttggtaggaa
     3241 tgttgagagg tattgccgct gggatgcgat acttggctga tatgggatac gttcacaggg
     3301 accttgcagc gcgcaacatc cttgtcaaca gcaatcttgt ttgtaaagtg tcagattttg
     3361 gcctttcccg ggttatagag gatgatcccg aagctgtcta caccacgact ggtggaaaaa
     3421 ttccagtaag gtggactgca ccggaagcca ttcaataccg gaagttcacc tcagccagcg
     3481 atgtgtggag ctatgggatt gtcatgtggg aagtgatgtc ttatggagaa agaccttact
     3541 gggacatgtc aaatcaagat gtcattaaag cgatagaaga aggttatcgt ttgccggcgc
     3601 ccatggattg cccagctggt cttcaccagc taatgctgga ttgttggcag aaagatcggg
     3661 cggaaaggcc aaagtttgag cagatagtcg gaattctaga caaaatgatt cgaaacccaa
     3721 gtagtctgaa aacacccctg ggaacttgta gtagaccctt aagccctctt ctggaccaga
     3781 gcactcctga cttcactgcc ttctgttcag ttggagaatg gttgcaagct attaaaatgg
     3841 aaaggtataa ggacaacttc acagcagcgg gttacaactc actcgagtca gtggccagga
     3901 tgactatcga tgatgtgatg agtttaggga tcacactggt tggccatcaa aagaagatca
     3961 tgagcagcat ccagactatg cgggcacaaa tgttgcattt acacggaaca ggcatccaag
     4021 tgacgcgtac gcggccgctc gagcagaaac tcatctcaga agaggatctg gcagcaaatg
     4081 atatcctgga ttacaaggat gacgacgata aggtttaaac ggccggccgc ggtcatagct
     4141 gtttcctgaa cagatcccgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc
     4201 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt
     4261 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag
     4321 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt
     4381 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg
     4441 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg
     4501 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct
     4561 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct
     4621 cccttccctg tccttctgat tttaaaataa ctataccagc aggaggacgt ccagacacag
     4681 cataggctac ctggccatgc ccaaccggtg ggacatttga gttgcttgct tggcactgtc
     4741 ctctcatgcg ttgggtccac tcagtagatg cctgttgaat tgggtacgcg gccagcggcg
     4801 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4861 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4921 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4981 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5041 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5101 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5161 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5221 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5281 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5341 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5401 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5461 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5521 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5581 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5641 aagttttaaa tcaatctaaa gtatatatga gtaacctgag gctatggcag ggcctgccgc
     5701 cccgacgttg gctgcgagcc ctgggccttc acccgaactt ggggggtggg gtggggaaaa
     5761 ggaagaaacg cgggcgtatt ggccccaatg gggtctcggt ggggtatcga cagagtgcca
     5821 gccctgggac cgaaccccgc gtttatgaac aaacgaccca acaccgtgcg ttttattctg
     5881 tctttttatt gccgtcatag cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt
     5941 tagcctcccc ctagggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat
     6001 ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga
     6061 atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt
     6121 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
     6181 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
     6241 tcacgggtag ccaacgctat gtcctgatag cgatccgcca cacccagccg gccacagtcg
     6301 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
     6361 gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa cagttcggct
     6421 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
     6481 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
     6541 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
     6601 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
     6661 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
     6721 agccgcgctg cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
     6781 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
     6841 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
     6901 aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc gatctttgca
     6961 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc
     7021 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
     7081 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
     7141 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
     7201 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     7261 ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc aggactcttc
     7321 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7381 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7441 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     7501 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     7561 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     7621 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     7681 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     7741 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     7801 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     7861 tttaacgcga atttt
//
