LOCUS       V031219                 8574 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031219
VERSION     V031219
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..4721
                     /label="Irs1(NM_010570)"
                     /note="Irs1(NM_010570)"
                     /gene="Irs1"
     CDS             4740..4769
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             4788..4811
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(4831..4847)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    4858..5480
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(5629..6217)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    6546..6593
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(6825..7619)
                     /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(7654..8011)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(8013..8117)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      8144..8572
                     /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 ggcgagccct ccggataccg atggcttctc agacgtgcgc aaggtgggct
     1081 acctgcgcaa gcccaagagt atgcataagc gctttttcgt gctgcgggcg gccagcgagg
     1141 ccgggggccc agcgcgcctg gagtattatg agaacgagaa gaagtggcgg cacaagtcga
     1201 gcgcccccaa acgctcgatc cccctcgaga gctgtttcaa catcaacaag cgggctgact
     1261 ccaagaacaa gcacctggtg gctctctaca cccgagacga acactttgcc attgccgcgg
     1321 atagcgaggc tgagcaagac agctggtacc aggctcttct gcagctgcat aatcgggcaa
     1381 aggcccacca tgacggggct ggaggaggct gcggcggtag ctgcagcggc agctctggcg
     1441 ttggagaggc aggggaggac ttgagctatg acacgggtcc aggacccgcg ttcaaggagg
     1501 tctggcaggt tatcctgaaa cccaagggcc tgggtcagac aaagaacctg attggcatct
     1561 accgcctctg cctgaccagc aagaccatca gctttgtgaa gctgaactcc gaggcagccg
     1621 ctgtggtgct gcagctgatg aatatcagac gctgtggcca ctcagagaac ttcttcttca
     1681 tcgaggtggg gcgttccgct gtgacagggc ctggcgagtt ctggatgcaa gtggatgact
     1741 ccgtggtggc ccagaacatg catgagacca ttctagaggc aatgagggcc atgagcgatg
     1801 agtttcgccc tcgcagcaaa agccagtctt catccagttg ctccaacccc atcagcgttc
     1861 ccctgcgcag gcaccatctc aacaaccctc cacccagcca ggtgggactg actcggagat
     1921 ctcgaactga gagcatcact gccacctccc ctgccagtat ggtgggtggg aaaccaggtt
     1981 ccttccgggt gcgtgcctcc agcgatggcg aaggcaccat gtcccgtcca gcatcagtgg
     2041 atggcagtcc tgtgagccct agcaccaaca ggacccacgc ccatcggcat cgaggcagct
     2101 ccaggctgca ccccccactc aaccacagcc gctccatccc catgccttct tctagatgct
     2161 caccttcagc caccagccca gtgagtctgt catctagtag taccagtggc catggctcca
     2221 cttcagactg tctcttcccg aggcgctcta gtgcttccgt gtccggttcg cctagcgatg
     2281 gcggtttcat ctcttctgat gagtatggtt ctagtccctg cgatttccga agttccttcc
     2341 gcagtgtcac cccagattcc ctggggcaca ccccaccagc caggggtgag gaggagctga
     2401 gcaattatat ctgcatgggt ggcaagggag cctccacctt ggctgctccc aatggccact
     2461 acattttgtc taggggtggc aacggccatc gctacatccc aggtgctaac ttggggacaa
     2521 gcccagcgct gcctggagat gaagccgcgg gtgcagcaga tctggataac cggtttcgaa
     2581 agagaactca ctccgcaggc acatctccta ccatttccca tcagaagacc ccctcacagt
     2641 cttcagtggc ttctattgag gaatatacag agatgatgcc cgctgcctac ccaccaggag
     2701 gtggcagtgg aggccgactg cccggctacc ggcattccgc cttcgtgccc acccactcct
     2761 atcccgaaga gggtctagag atgcaccact tggaacgtcg tggaggccac caccgtccag
     2821 acacctccaa cctccacact gatgatggct atatgcccat gtctcctggg gtggctccag
     2881 tgcccagcaa ccgcaaagga aatggggact atatgcccat gagccccaag agtgtatctg
     2941 ccccacagca gatcattaac cccatcagac gccacccaca gagagtggac cccaatggct
     3001 acatgatgat gtcacccagt ggtagttgct cccctgacat tggaggtggg tccagcagca
     3061 gtagcagcat cagcgcagcc ccttctggga gcagctatgg gaagccatgg acaaatggag
     3121 taggggggca ccatactcat gcccttcctc atgccaaacc tcctgttgag agtggtggcg
     3181 gtaagctctt gccttgcaca ggtgactaca tgaacatgtc cccagtggga gattccaaca
     3241 ccagcagccc ctcagaatgc tactatggcc cagaagatcc ccagcacaag ccggtcctct
     3301 cttactactc attgccaagg tcctttaagc acacccagcg ccctggagag ccagaggagg
     3361 gtgccaggca ccagcatctt cgtctctctt ctagctctgg acgccttcgc tataccgcaa
     3421 ctgccgaaga ttcctcctct tctaccagca gcgacagtct gggtgggggt tactgtgggg
     3481 ctaggccaga gtcaagcctc acacatcccc accaccatgt cttgcagccc cacctgcctc
     3541 gaaaggtaga cacagctgca cagaccaaca gccgcctggc tcgacccaca aggctgtcct
     3601 tgggggatcc caaggcaagc accttacccc gggttcgaga gcagcaacag cagcagcagt
     3661 cttccctgca ccctcccgag cccaaaagcc caggagaata tgtgaatatt gaattcggga
     3721 gcggccagcc tggctattta gctggccctg caacttcccg tagctctcct tcagttcgat
     3781 gtccacccca gctccaccca gctcctagag aagagactgg ctcggaagag tacatgaaca
     3841 tggacttggg gccaggccgg agggcaacct ggcaggagag tggtggagtt gagttgggca
     3901 gaataggccc tgcacctccg gggtctgcta cggtttgcag gccaacccgt tcggtgccaa
     3961 atagccgtgg tgactacatg accatgcaga taggttgtcc tcgtcaaagc tatgtggata
     4021 cctcaccagt ggccccagtc agctatgctg acatgcggac aggcattgct gcagagaagg
     4081 cgagcctgcc tagacccaca ggagctgctc ctcctccatc ctccacagcc tcttcttctg
     4141 ttacacctca aggagccacc gctgagcaag ctactcactc ttccttgctg ggaggccctc
     4201 agggacctgg gggcatgagt gcattcacca gggtgaacct cagtcccaac cataaccaga
     4261 gtgccaaagt gattcgcgca gacactcaag ggtgccggag gaggcatagc tctgagacct
     4321 tctcagcacc tactcgggct ggcaatacgg tgccctttgg agcaggggct gcagtagggg
     4381 gcagcggcgg tggtggtggt ggcggcagtg aggatgtaaa acgccacagc tctgcatcct
     4441 ttgagaatgt gtggctgaga cctggggatc tagggggagt ctccaaggag tcggctccag
     4501 tgtgtggggc tgctgggggt ttggagaaga gtcttaacta catagacctg gatttggcca
     4561 aggagcgctc tcaggactgc ccctctcaac agcagtccct accaccccct ccccctcacc
     4621 agcccttagg cagcaatgag ggcaactccc caagacgctc cagtgaggat ttaagcaact
     4681 atgccagcat cagcttccag aagcagccag aggatcgtca aacgcgtacg cggccgctcg
     4741 agcagaaact catctcagaa gaggatctgg cagcaaatga tatcctggat tacaaggatg
     4801 acgacgataa ggtttaaacg gccggccgcg gtcatagctg tttcctgaac agatcccggg
     4861 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     4921 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     4981 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     5041 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     5101 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
     5161 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     5221 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     5281 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
     5341 ttaaaataac tataccagca ggaggacgtc cagacacagc ataggctacc tggccatgcc
     5401 caaccggtgg gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact
     5461 cagtagatgc ctgttgaatt gggtacgcgg ccagcggcga gcggtatcag ctcactcaaa
     5521 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5581 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5641 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5701 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5761 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5821 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5881 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5941 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6001 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6061 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6121 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6181 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6241 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6301 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6361 tatatatgag taacctgagg ctatggcagg gcctgccgcc ccgacgttgg ctgcgagccc
     6421 tgggccttca cccgaacttg gggggtgggg tggggaaaag gaagaaacgc gggcgtattg
     6481 gccccaatgg ggtctcggtg gggtatcgac agagtgccag ccctgggacc gaaccccgcg
     6541 tttatgaaca aacgacccaa caccgtgcgt tttattctgt ctttttattg ccgtcatagc
     6601 gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc tagggtgggc
     6661 gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac
     6721 gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag
     6781 gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca
     6841 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
     6901 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
     6961 tcctgatagc gatccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
     7021 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
     7081 tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
     7141 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
     7201 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
     7261 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     7321 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     7381 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc
     7441 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
     7501 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
     7561 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg
     7621 cgaaacgatc ctcatcctgt ctcttgatcg atctttgcaa aagcctaggc ctccaaaaaa
     7681 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat
     7741 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
     7801 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
     7861 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
     7921 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
     7981 cacattccac agctggttct ttccgcctca ggactcttcc tttttcaata ttattgaagc
     8041 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8101 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc
     8161 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     8221 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     8281 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     8341 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     8401 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     8461 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     8521 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa tttt
//
