LOCUS       V030436                 8490 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030436
VERSION     V030436
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..4637
                     /label="Grm5(NM_001143834)"
                     /note="Grm5(NM_001143834)"
                     /gene="Grm5"
     CDS             4656..4685
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             4704..4727
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(4747..4763)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    4774..5396
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(5545..6133)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    6462..6509
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(6741..7535)
                     /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(7570..7927)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(7929..8033)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      8060..8488
                     /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 ggtccttctg ttgattctgt cagtcctact tctgaaagaa gatgtacgag
     1081 gaagtgcaca gtccagtgag aggagggtgg tggctcacat gccaggtgac attattattg
     1141 gagctctctt ctcggtccac caccaaccaa ctgtggacaa agttcatgag aggaagtgtg
     1201 gggcagtccg tgagcagtat ggcattcaga gagtggaagc catgctgcat accttggaaa
     1261 ggatcaattc agatcccaca ctcttgccca acatcacact aggctgtgag ataagagatt
     1321 cctgctggca ttctgctgtg gccctagagc aaagcattga gtttataagg gattccctca
     1381 tctcttcgga agaggaagaa ggcttggtac gctgtgtaga tggctcttct tccttccgct
     1441 ccaagaaacc catagtggga gtcattgggc ctggctcgag ttctgtggcc attcaagttc
     1501 agaacttgct ccagcttttc aacatacctc agattgctta ctctgcaact agcatggatt
     1561 tgagtgacaa gactctattc aagtacttca tgagggttgt accttcggat gcccagcaag
     1621 cccgagccat ggtagacata gtgaagagat acaactggac ttatgtctca gctgtgcaca
     1681 cagaaggcaa ctatggagaa agtgggatgg aggctttcaa agatatgtca gcgaaggaag
     1741 ggatttgcat cgcccactct tacaaaatct acagcaatgc tggggaacag agctttgaca
     1801 agctgttgaa aaagctcaga agtcatttac ctaaagcccg ggtggtagcc tgcttctgtg
     1861 aaggcatgac agttcgaggt ctgctcatgg ccatgagacg cttgggtcta gcaggggaat
     1921 ttctacttct gggcagtgat ggctgggctg acaggtatga cgtgacagat gggtatcagc
     1981 gagaagctgt cggtgggatt acaatcaagc tccagtctcc tgatgtcaag tggtttgatg
     2041 attattatct gaagctccgg ccagaaacaa acctcagaaa cccttggttt caagaatttt
     2101 ggcagcatcg ttttcagtgc cggctagaag ggtttgcaca ggagaacagc aagtacaaca
     2161 agacttgcaa cagttctcta actctgagaa cgcatcatgt tcaagattcc aaaatgggat
     2221 ttgtgatcaa tgcaatctat tctatggctt atgggctcca caacatgcag atgtccctgt
     2281 gtccaggcta tgcaggcctc tgtgatgcaa tgaagccaat tgatgggcgg aaacttttgg
     2341 actccctgat gaaaaccaac tttactggag tttccggaga tatgattcta tttgatgaaa
     2401 atggagactc tccaggaagg tatgaaataa tgaatttcaa ggaaatggga aaagattatt
     2461 ttgattacat caatgttgga agttgggaca atggggaatt aaaaatggat gatgatgaag
     2521 tgtggtccaa gaaaaataac atcatcagat ctgtgtgcag tgaaccgtgt gagaaaggac
     2581 agataaaggt gatccggaag ggagaagtca gctgttgttg gacctgcaca ccttgtaagg
     2641 agaatgagta tgtttttgat gagtacacct gcaaggcgtg ccaactgggg tcctggccca
     2701 ctgacgactt gacaggttgt gatttgatcc cagtccagta tcttcgatgg ggtgaccctg
     2761 agcccattgc agctgtggtg tttgcctgcc tcggtctgct agccaccctc ttcgttactg
     2821 taatcttcat catttatcgg gacactccag tggtcaagtc ttccagcagg gaactctgct
     2881 acattatcct tgctggcatc tgcctgggtt acttgtgtac cttctgcctc attgcaaagc
     2941 ccaaacagat ttactgctac cttcagagaa ttggcatcgg tctctctcca gccatgagct
     3001 actcagccct tgtaacgaag accaaccgta ttgcaaggat tctagctggc agcaagaaga
     3061 agatctgtac caaaaagccc agattcatga gcgcctgtgc tcagttagtg atcgctttca
     3121 ttctcatctg tatacagttg ggtattattg tggccctctt tatcatggag cctccggata
     3181 taatgcatga ctatccaagc atccgagaag tctacttgat ttgtaacacc accaacctag
     3241 gggttgtcac tcctcttgga tacaatggat tattgatttt gagttgcaca ttctatgcgt
     3301 tcaagaccag aaatgttcca gccaacttta acgaggccaa atatattgct ttcaccatgt
     3361 acacaacctg catcatatgg ctggcctttg tgcctatcta ctttggcagc aactacaaaa
     3421 tcatcaccat gtgtttctca gtcagcctca gtgccacagt ggccctgggt tgcatgtttg
     3481 tgccgaaggt gtacatcatc ctagccaaac cggagagaaa tgtgcgcagc gccttcacaa
     3541 cctctacagt ggtgcgcatg cacgtaggag atggcaagtc atcatccgct gccagcagat
     3601 ccagcagcct agtcaacctg tggaagagga ggggctcgtc tggggaaacc ctaaggtaca
     3661 aagacaggag actggcccag cacaagtcgg aaatagagtg tttcaccccc aaagggagta
     3721 tggggaatgg tgggagagca acaatgagca gctccaatgg aaaatccgtg acttgggccc
     3781 agaatgagaa gagcacccgg gggcagcacc tgtggcagcg actgtctgtc catatcaaca
     3841 agaaggagaa ccccaaccag acagcagtca tcaaaccctt ccccaagagc acagagagcc
     3901 gcgggcaggg tgcaggggca ggtggtggct ctggccccgg tgcagctggt gctggtagcg
     3961 caggatgcac agcgacaggc ggcccagagc caccagacgc cggccccaag gcgctttatg
     4021 atgtcgcaga ggcagaggag cgcttcccag cggctgccag gccgcgctcg ccatcgccca
     4081 tcagtacgct gagccacctg gcaggctcgg cgggccgcac agacgacgac gcgccgtcgc
     4141 tgcactcgga gaccgctgca cgcagcagct catcccaggg ctcgctcatg gagcagatta
     4201 gcagcgtggt gacgcgcttc accgccaaca tcaccgagct caactccatg atgttgtcca
     4261 ccgcggctgc gccggggccc cctggtaccc ctatctgctc ttcctacctg atccccaaag
     4321 agatccagct gcccacgacc atgacgacct tcgcagagat ccagcctctg ccggccatcg
     4381 aggtgaccgg aggagctcag ccggcgacag ggccatcacc tgcccaagag acgcccgcag
     4441 gagctgaagc cgccccagga aaaccggatc tggaggagct ggtggccctc actccaccat
     4501 cgcccttcag ggactcggtg gactcgggga gcaccacccc aaactctcca gtctccgaat
     4561 cggccctctg catcccatcc tctcccaaat atgacactct catcatcaga gattacacgc
     4621 agagttcttc atcgttgacg cgtacgcggc cgctcgagca gaaactcatc tcagaagagg
     4681 atctggcagc aaatgatatc ctggattaca aggatgacga cgataaggtt taaacggccg
     4741 gccgcggtca tagctgtttc ctgaacagat cccgggtggc atccctgtga cccctcccca
     4801 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa
     4861 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg
     4921 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg
     4981 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt
     5041 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg
     5101 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc
     5161 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag
     5221 gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa aataactata ccagcaggag
     5281 gacgtccaga cacagcatag gctacctggc catgcccaac cggtgggaca tttgagttgc
     5341 ttgcttggca ctgtcctctc atgcgttggg tccactcagt agatgcctgt tgaattgggt
     5401 acgcggccag cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5461 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5521 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5581 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5641 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5701 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5761 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5821 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5881 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5941 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6001 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6061 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6121 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6181 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6241 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac ctgaggctat
     6301 ggcagggcct gccgccccga cgttggctgc gagccctggg ccttcacccg aacttggggg
     6361 gtggggtggg gaaaaggaag aaacgcgggc gtattggccc caatggggtc tcggtggggt
     6421 atcgacagag tgccagccct gggaccgaac cccgcgttta tgaacaaacg acccaacacc
     6481 gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc
     6541 cttccgtgtt tcagttagcc tccccctagg gtgggcgaag aactccagca tgagatcccc
     6601 gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata
     6661 gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat
     6721 ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc
     6781 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca
     6841 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcgatc cgccacaccc
     6901 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag
     6961 caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg
     7021 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca
     7081 agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat
     7141 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact
     7201 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc
     7261 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc
     7321 gtggccagcc acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg
     7381 tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca
     7441 gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc
     7501 ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct
     7561 tgatcgatct ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     7621 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     7681 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     7741 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     7801 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     7861 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct ggttctttcc
     7921 gcctcaggac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7981 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     8041 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     8101 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     8161 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     8221 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     8281 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     8341 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     8401 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     8461 ctgatttaac aaaaatttaa cgcgaatttt
//
