LOCUS       V031234                 8844 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031234
VERSION     V031234
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..4991
                     /label="Irs2(NM_001081212)"
                     /note="Irs2(NM_001081212)"
                     /gene="Irs2"
     CDS             5010..5039
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             5058..5081
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(5101..5117)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    5128..5750
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(5899..6487)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    6816..6863
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(7095..7889)
                     /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(7924..8281)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(8283..8387)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      8414..8842
                     /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 ggctagcgcg cccctgcctg ggccccccgc gtcggcgggc ggggacggcc
     1081 cgaacctcaa taacaacaac aacaacaaca accacagcgt gcgcaagtgc ggctacctgc
     1141 gcaagcagaa gcacggccac aagcgctttt tcgtgttgcg cggccccggc acgggcggcg
     1201 acgaggcatc cgcggctggg gggtcgccgc cgcagcctcc gcggctggag tactacgaga
     1261 gcgagaagaa gtggaggagc aaggcgggcg cgccgaagcg agtgatcgcg ctcgactgct
     1321 gtctgaacat caacaagcgc gcggacgcca agcacaagta cctgatcgcc ctctacacca
     1381 aggacgagta cttcgctgta gcggcggaga acgagcagga gcaggagggc tggtaccgcg
     1441 cactcaccga cttggtcagc gaaggccgct ctggcgaggg gggctcgggc accaccggag
     1501 gctcttgcag cgcctctctc ccgggcgtcc tgggcggctc agcgggcgcc gctggctgcg
     1561 atgacaacta cgggctcgtg acacccgcca cggccgtcta ccgcgaggtg tggcaggtga
     1621 acctgaaacc taagggactg ggccagagca agaacctgac tggtgtatac cgcctatgcc
     1681 tgtctgcgcg caccatcggc ttcgtgaagc tcaattgcga acagccgtcg gtgacgctgc
     1741 agcttatgaa cattcgccgc tgcggccact cggacagctt cttcttcatc gaggtgggcc
     1801 gttcggccgt caccggtccc ggggagctgt ggatgcaagc cgacgactcg gtggtggcgc
     1861 agaacatcca tgagaccatc ctagaagcta tgaaggcact caaagagctc ttcgagttcc
     1921 ggcctcgcag caagagtcag tcgtccgggt cgtcagccac gcatcccatc agcgtgccgg
     1981 gcgcgcgccg ccaccaccac ctagtcaacc taccccctag ccagaccggc ctggtgcgcc
     2041 gctcgcgcac tgacagcctg gcggccaccc ccccagcagc caagtgcact tcgtgccggg
     2101 ttcgtacggc cagcgagggc gacggcggcg cggcaggcgg ggccgggacg gcaggaggca
     2161 ggccgatgtc ggtggcaggg agccccctga gtcccgggcc ggtgcgcgcg ccccttagcc
     2221 gctcgcacac cctgagcgcc ggctgcggag gccgcccgag caaagtgact ctggcgccgg
     2281 cagggggagc cctgcaacac agccgctcca tgtccatgcc cgtggcgcac tcacctcctg
     2341 cagccaccag cccaggcagc ctgtcctcca gcagtgggca cggctcgggc tcctacccgc
     2401 tgccacctgg ctcccacccg cacctgcctc atccactgca tcacccccaa ggccagcgtc
     2461 cgtccagcgg tagtgcctcc gcctcgggct cccccagcga cccgggtttc atgtcccttg
     2521 acgagtatgg ctccagccct ggcgacctga gagccttcag tagccacagg agcaacacac
     2581 ccgagtcaat agcggagacc ccgccagcca gagatggcag tgggggcgaa ctctatgggt
     2641 acatgagcat ggatagaccc ctgagccact gtggccgccc ttaccgtagg gtctcagggg
     2701 atggggccca ggacctggat agaggactga ggaagaggac ttattcccta accacgcctg
     2761 ccaggcagcg gcaggtacct cagccttcct ctgcctctct agatgaatac actctcatga
     2821 gggccacctt ctctggtagt tcaggtcgcc tctgcccatc cttccctgcg tcctctccca
     2881 aagtggccta caacccttac ccagaggact atggagacat tgagattggt tctcacaaga
     2941 gttccagcag taacctgggg gcagatgatg gctacatgcc catgacccct ggggcagccc
     3001 ttaggagtgg tggtcccaat agctgcaaga gcgatgacta catgcccatg agccccacaa
     3061 gcgtgtctgc tcccaagcag atcctgcagc cacgcttggc agcggccttg cccccttccg
     3121 gagcagccgt gccagcaccc ccttcagggg tgggcaggac cttcccagta aacggaggtg
     3181 gctacaaagc cagctcccca gcggagagct ccccagaaga cagtgggtac atgcgaatgt
     3241 ggtgtggctc caagctgtct atggagaacc cagaccctaa gctactcccc aacggggact
     3301 acctcaacat gtcccccagc gaggcaggca ctgcagggac cccacctgac ttctcagcag
     3361 ctttgcgtgg aggcagtgaa ggcctcaaag gcatcccggg ccactgctac agctctttgc
     3421 cccgctctta taaggctccc tgttcctgca gcggagacaa tgaccagtat gtgctcatga
     3481 gctcccctgt gggccggatc ttggaagagg agagactgga gccccaggcc accccagggg
     3541 ctggcacctt tggggcagct ggtggtagtc atacccagcc tcatcactca gcagtgcctt
     3601 cctccatgag gccgagtgcc atcggtggcc gccctgaggg cttcctgggc cagcgatgtc
     3661 gggcagtgcg gcctacacgc ctatcgctag agggactgca gacccttccc agcatgcaag
     3721 agtaccctct acccacagag cccaagagcc ctggcgagta catcaacatt gactttggtg
     3781 aggcaggtac ccgtctgtct ccgcctgccc ccccactact ggcatccgcg gcctcatctt
     3841 cttcactgct ctcagctagt agtcctgctt catccctggg ttcaggaacc ccaggcacca
     3901 gcagcgacag ccggcagcgc tctccactct ctgactatat gaacctggac ttcagttctc
     3961 ccaagtcccc caagcctagc acccgcagtg gggacacagt aggctccatg gatggccttc
     4021 tctctccaga ggcttcatcc ccatacccac cactgccccc acgtccttcc acttcccctt
     4081 cctccttaca gcagcctctg ccacctgccc cgggagacct ataccgcctg cctccagcat
     4141 cagctgccac ttcccagggt cccactgctg gctcctcaat gtcctccgag cctggggata
     4201 atggtgacta taccgagatg gcctttggtg tggctgcaac cccgccacaa cctatcgtgg
     4261 cacctccaaa gccagaaggt gcccgagtgg ccagtcccac atcgggcttg aagcggctaa
     4321 gtctcatgga tcaggtatct ggggtggagg ctttccttca agtcagccag ccccctgacc
     4381 cccaccgggg tgctaaggtc atccgtgcag acccacaggg gggacgtcgt cgccacagtt
     4441 cagagacctt ttcctctacc accaccgtca ccccagtgtc cccatccttt gcccacaatt
     4501 ccaagcgcca caattcggcc tctgtggaaa atgtctcact caggaaaagc agtgaaggca
     4561 gcagtaccct gggaggaggt gatgagccgc ccacatcccc aggacaggca cagcccttgg
     4621 tggctgtgcc cccagtgcca caggctaggc cgtggaaccc cggtcagccc ggagctttga
     4681 ttggctgtcc tggaggcagc agttctccca tgcgcagaga gacctccgtg ggtttccaga
     4741 acggcctcaa ctatatcgcc atcgatgtga gaggcgagca ggggtccttg gcgcagtctc
     4801 agccgcagcc aggagacaag aactcctgga gccggacccg tagccttggg gggctcctcg
     4861 gcaccgtcgg aggctctggc gccagcggag tgtgtggggg tccaggcact ggagctttgc
     4921 cctctgccag cacctatgca agcatcgact tcctgtccca tcacttgaag gaagccacag
     4981 tcgtgaaaga gacgcgtacg cggccgctcg agcagaaact catctcagaa gaggatctgg
     5041 cagcaaatga tatcctggat tacaaggatg acgacgataa ggtttaaacg gccggccgcg
     5101 gtcatagctg tttcctgaac agatcccggg tggcatccct gtgacccctc cccagtgcct
     5161 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
     5221 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
     5281 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
     5341 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
     5401 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
     5461 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
     5521 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
     5581 accactgctc ccttccctgt ccttctgatt ttaaaataac tataccagca ggaggacgtc
     5641 cagacacagc ataggctacc tggccatgcc caaccggtgg gacatttgag ttgcttgctt
     5701 ggcactgtcc tctcatgcgt tgggtccact cagtagatgc ctgttgaatt gggtacgcgg
     5761 ccagcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5821 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5881 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5941 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6001 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6061 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6121 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6181 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6241 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6301 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6361 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6421 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6481 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6541 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6601 ttaaaaatga agttttaaat caatctaaag tatatatgag taacctgagg ctatggcagg
     6661 gcctgccgcc ccgacgttgg ctgcgagccc tgggccttca cccgaacttg gggggtgggg
     6721 tggggaaaag gaagaaacgc gggcgtattg gccccaatgg ggtctcggtg gggtatcgac
     6781 agagtgccag ccctgggacc gaaccccgcg tttatgaaca aacgacccaa caccgtgcgt
     6841 tttattctgt ctttttattg ccgtcatagc gcgggttcct tccggtattg tctccttccg
     6901 tgtttcagtt agcctccccc tagggtgggc gaagaactcc agcatgagat ccccgcgctg
     6961 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc
     7021 ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa
     7081 ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa
     7141 tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct
     7201 tcagcaatat cacgggtagc caacgctatg tcctgatagc gatccgccac acccagccgg
     7261 ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca
     7321 tcgccatggg tcacgacgag atcctcgccg tcgggcatgc tcgccttgag cctggcgaac
     7381 agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg
     7441 gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag
     7501 gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg
     7561 gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag
     7621 tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc
     7681 agccacgata gccgcgctgc ctcgtcttgc agttcattca gggcaccgga caggtcggtc
     7741 ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag
     7801 ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa
     7861 cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatcg
     7921 atctttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga
     7981 ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga
     8041 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
     8101 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     8161 tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     8221 gcctggggac tttccacacc ctaactgaca cacattccac agctggttct ttccgcctca
     8281 ggactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     8341 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     8401 aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     8461 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     8521 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     8581 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     8641 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     8701 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     8761 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     8821 taacaaaaat ttaacgcgaa tttt
//
