LOCUS       V030461                 9096 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030461
VERSION     V030461
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..5243
                     /label="Crb1(NM_133239)"
                     /note="Crb1(NM_133239)"
                     /gene="Crb1"
     CDS             5262..5291
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             5310..5333
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(5353..5369)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    5380..6002
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(6151..6739)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    7068..7115
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(7347..8141)
                     /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(8176..8533)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(8535..8639)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      8666..9094
                     /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 gaagctcaag agaactgcct accttctctt cctgtacctc agctcctcac
     1081 tgctcatctg cataaagaat tcattttgca ataaaaacaa taccaggtgc ctttcaggtc
     1141 cttgccaaaa caattctacg tgcaagcatt ttccacaaga caacaattgt tgcttagaca
     1201 cagccaataa tttggacaaa gactgtgaag atctgaaaga cccttgcttc tcgagtccct
     1261 gccaaggaat tgccacttgt gtgaaaatcc caggggaagg gaacttcctg tgtcagtgtc
     1321 ctcctgggta cagcgggctg aactgtgaaa ctgccaccaa ttcctgtgga gggaacctct
     1381 gccaacatgg aggcacctgc cgtaaagacc ctgagcaccc tgtctgtatc tgccctcctg
     1441 gatatgctgg aaggttctgt gagactgatc acaatgagtg tgcttctagc ccttgccaca
     1501 atggggctat gtgccaggat ggaatcaatg gctactcctg cttctgtgtg cctggatacc
     1561 aaggcaggca ttgtgacttg gaagtggatg aatgtgtttc tgatccctgc aagaatgagg
     1621 ctgtgtgcct caatgagata ggaagataca cttgtgtctg ccctcaagag ttttctggcg
     1681 tgaactgtga gttggaaatt gatgaatgca gatcccagcc ttgtctccac ggtgccacat
     1741 gtcaggacgc tccagggggc tactcctgtg actgtgcacc tggattcctt ggagagcact
     1801 gtgaactcag cgttaatgaa tgtgaaagtc agccgtgtct ccatggaggt ctatgtgtgg
     1861 atggaagaaa cagttaccac tgtgactgca caggtagtgg attcacaggg atgcactgtg
     1921 agtccttgat tcctctttgt tggtcaaagc cttgtcacaa cgacgcgaca tgtgaagata
     1981 ctgttgacag ctatatttgt cactgccggc ctggatacac aggtgccctg tgtgagacag
     2041 acataaatga atgcagtagc aacccctgcc aattttgggg ggaatgtgtc gagctgtcct
     2101 cagagggtct atatggaaac actgctggcc tgccttcctc cttcagctat gttggagcct
     2161 cgggctatgt gtgtatctgt cagcctggat tcacaggaat tcactgtgaa gaagacgttg
     2221 atgaatgttt actgcaccct tgcctaaatg gtggtacttg tgagaacctg cctgggaatt
     2281 atgcctgtca ctgtcccttt gatgacactt ctaggacatt ttatggagga gaaaactgct
     2341 cagaaattct cctgggctgc actcatcacc agtgtctgaa caatggaaaa tgtatccctc
     2401 atttccaaaa tggccagcat ggattcactt gccagtgtct ttctggctat gcggggcccc
     2461 tgtgtgaaac tgtcaccaca ctttcatttg ggagcaatgg cttcctatgg gtcacaagtg
     2521 gctcccatac aggcataggg ccagaatgta acatatcctt gaggtttcac actgttcaac
     2581 caaacgcact tctcctcatc cgaggcaaca aggacgtgtc tatgaagctg gagttgctga
     2641 atggttgtgt tcacttatca attgaagtct ggaatcagtt aaaggtgctc ctgtctattt
     2701 ctcacaacac cagtgatgga gaatggcatt tcgtggaggt aacaatcgca gaaactctaa
     2761 cccttgccct agttggcggc tcctgcaagg agaagtgcac caccaagtct tctgttccag
     2821 ttgagaatca tcaatcaata tgtgctttgc aggactcttt tttgggtggc ttaccaatgg
     2881 ggacagccaa caacagtgtg tctgtgctta acatctataa tgtgccgtcc acaccttcct
     2941 ttgtaggctg tctccaagac attagatttg atttgaatca cattactctg gagaacgttt
     3001 catctggcct gtcatcaaat gttaaagcag gctgcctggg aaaggactgg tgtgaaagtc
     3061 aaccctgtca aaacagagga cgctgcatca acttgtggca gggttatcag tgtgaatgtg
     3121 acaggcccta tacaggctcc aactgcctga aagagtatgt agcgggaaga tttggccaag
     3181 atgactccac aggatatgcg gcctttagtg ttaatgataa ttatggacag aacttcagtc
     3241 tttcaatgtt tgtccgaaca cgtcaacccc tgggcttact tctggctttg gaaaatagta
     3301 cttaccagta tgtcagtgtc tggctagagc acggcagcct agcactgcag actccaggct
     3361 ctcccaagtt catggtaaac ttttttctca gtgatggaaa tgttcactta atatctttga
     3421 gaatcaaacc aaatgaaatt gaactgtatc agtcttcaca aaacctagga ttcatttctg
     3481 ttcctacatg gacaattcga agaggagacg tcatcttcat tggtggctta cctgacagag
     3541 agaagactga agtttatggt ggcttcttca aaggctgtgt tcaagatgtc agattaaaca
     3601 gccagactct ggaattcttt cccaattcaa caaacaatgc atacgatgac ccaattcttg
     3661 tcaatgtgac tcaaggctgt cccggagaca acacatgtaa gtccaacccc tgtcataatg
     3721 gaggtgtctg ccactccctg tgggatgact tctcctgctc ctgccctaca aacacagcgg
     3781 ggagagcctg cgagcaagtt cagtggtgtc aactcagccc atgtcctccc actgcagagt
     3841 gccagctgct ccctcaaggg tttgaatgta tcgcaaacgc tgttttcagc ggattaagca
     3901 gagaaatact cttcagaagc aatgggaaca ttaccagaga actcaccaat atcacatttg
     3961 ctttcagaac acatgataca aatgtgatga tattgcatgc agaaaaagaa ccagagtttc
     4021 ttaatattag cattcaagat gccagattat tctttcaatt gcgaagtggc aacagctttt
     4081 atacgctgca cctgatgggt tcccaattgg tgaatgatgg cacatggcac caagtgactt
     4141 tctccatgat agacccagtg gcccagacct cccggtggca aatggaggtg aacgaccaga
     4201 caccctttgt gataagtgaa gttgctactg gaagcctgaa ctttttgaag gacaatacag
     4261 acatctatgt gggtgaccaa tctgttgaca atccgaaagg cctgcagggc tgtctgagca
     4321 caatagagat tggaggcata tatctttctt actttgaaaa tctacatggt ttccctggta
     4381 agcctcagga agagcaattt ctcaaagttt ctacaaatat ggtacttact ggctgtttgc
     4441 catcaaatgc ctgccactcc agcccctgtt tgcatggagg aaactgtgaa gacagctaca
     4501 gttcttatcg gtgtgcctgt ctctcgggat ggtcagggac acactgtgaa atcaacattg
     4561 atgagtgctt ttctagcccc tgtatccatg gcaactgctc tgatggagtt gcagcctacc
     4621 actgcaggtg tgagcctgga tacaccggtg tgaactgtga ggtggatgta gacaattgca
     4681 agagtcatca gtgtgcaaat ggggccacct gtgttcctga agctcatggc tactcttgtc
     4741 tctgctttgg aaattttacc gggagatttt gcagacacag cagattaccc tcaacagtct
     4801 gtgggaatga gaagagaaac ttcacttgct acaatggagg cagctgctcc atgttccagg
     4861 aggactggca atgtatgtgc tggccaggtt tcactggaga gtggtgtgaa gaggacatca
     4921 acgagtgtgc ctccgatccc tgcatcaatg gaggactgtg cagggacttg gtcaacaggt
     4981 tcctatgcat ctgtgatgtg gccttcgctg gcgagcgctg tgagctggac ctggctgatg
     5041 acaggctcct gggcattttc accgctgttg gctccggaac tttggccctg ttcttcatcc
     5101 tcttgcttgc tggggttgct tctcttattg cctccaacaa aagggcgact caaggaacct
     5161 acagccccag cggtcaggag aaggctggcc ctcgagtgga aatgtggatc aggatgccgc
     5221 ccccggcact ggaaaggctc atcacgcgta cgcggccgct cgagcagaaa ctcatctcag
     5281 aagaggatct ggcagcaaat gatatcctgg attacaagga tgacgacgat aaggtttaaa
     5341 cggccggccg cggtcatagc tgtttcctga acagatcccg ggtggcatcc ctgtgacccc
     5401 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta
     5461 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg
     5521 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc
     5581 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc
     5641 tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg
     5701 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg
     5761 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga
     5821 ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttaaaata actataccag
     5881 caggaggacg tccagacaca gcataggcta cctggccatg cccaaccggt gggacatttg
     5941 agttgcttgc ttggcactgt cctctcatgc gttgggtcca ctcagtagat gcctgttgaa
     6001 ttgggtacgc ggccagcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6061 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6121 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6181 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6241 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6301 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6361 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6421 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6481 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6541 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6601 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6661 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6721 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6781 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6841 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaacctga
     6901 ggctatggca gggcctgccg ccccgacgtt ggctgcgagc cctgggcctt cacccgaact
     6961 tggggggtgg ggtggggaaa aggaagaaac gcgggcgtat tggccccaat ggggtctcgg
     7021 tggggtatcg acagagtgcc agccctggga ccgaaccccg cgtttatgaa caaacgaccc
     7081 aacaccgtgc gttttattct gtctttttat tgccgtcata gcgcgggttc cttccggtat
     7141 tgtctccttc cgtgtttcag ttagcctccc cctagggtgg gcgaagaact ccagcatgag
     7201 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
     7261 ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc
     7321 ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg
     7381 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
     7441 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcgatccgcc
     7501 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
     7561 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg
     7621 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     7681 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     7741 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     7801 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     7861 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     7921 cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg
     7981 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     8041 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     8101 gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct
     8161 gtctcttgat cgatctttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     8221 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
     8281 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
     8341 cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     8401 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct
     8461 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt
     8521 ctttccgcct caggactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8581 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8641 acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt
     8701 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     8761 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     8821 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     8881 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     8941 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     9001 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     9061 aatgagctga tttaacaaaa atttaacgcg aatttt
//
