LOCUS       V031183                 8187 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031183
VERSION     V031183
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..4334
                     /label="Pik3cg(NM_001146200)"
                     /note="Pik3cg(NM_001146200)"
                     /gene="Pik3cg"
     CDS             4353..4382
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             4401..4424
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(4444..4460)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    4471..5093
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(5242..5830)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    6159..6206
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(6438..7232)
                     /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(7267..7624)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(7626..7730)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      7757..8185
                     /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 ggagctggag aactatgaac aaccggtggt tctaagagag gacaacctcc
     1081 gccggcgccg gaggatgaag ccacgcagcg cagcaggcag cctgtcttcc atggagctca
     1141 tccccattga gttcgtactg cccaccagcc agcgcatcag caagactcca gaaacagcgc
     1201 tgctgcatgt ggctggccat ggcaatgtgg aacagatgaa agctcaggtg tggctgcgcg
     1261 cactggagac cagtgtggct gcggagttct accaccgatt gggcccggac caattcctcc
     1321 tgctctacca gaagaaagga caatggtatg agatctatga caggtaccaa gtggtgcaga
     1381 ccctagactg cctgcattac tggaagttga tgcacaagag ccctggccag atccacgtgg
     1441 tacagcgaca cgtaccttct gaggagacct tggctttcca gaagcagctc acctccctga
     1501 ttggctatga cgtcactgac atcagcaatg tgcacgatga tgagctagag ttcactcgcc
     1561 gccgtctggt tacgccccgc atggctgaag tggctggccg ggatgccaaa ctctatgcta
     1621 tgcacccttg ggtaacgtcc aaacctctcc cagactacct gtcaaaaaag attgccaaca
     1681 actgcatctt catcgtcatc caccgcggta ccaccagcca aaccatcaag gtctccgcag
     1741 atgatactcc tggtaccatc ctccagagct tcttcaccaa gatggccaag aagaagtccc
     1801 taatgaatat ctcagaaagt caaagtgagc aggattttgt attgcgggtt tgtggccgcg
     1861 atgagtacct ggtgggtgaa acacccctca aaaatttcca gtgggtgagg cagtgcctca
     1921 agaacggaga tgaaatacac ctggtgctcg acacgcctcc agacccagcc cttgatgagg
     1981 tgaggaagga agaatggccg ctggtggatg actgcactgg agtcaccggc taccacgagc
     2041 agctgaccat ccatggcaag gaccacgaga gtgtgttcac agtgtctttg tgggactgcg
     2101 accgaaagtt cagggtcaag atcagaggca ttgatatccc tgtcctgcct cggaacaccg
     2161 acctcactgt gtttgtggaa gcgaacatcc agcacgggca acaagtcctc tgccaaagga
     2221 gaaccagccc taagcccttc gcagaagagg tactctggaa tgtgtggctg gagtttggca
     2281 tcaaaatcaa agacttgccc aaaggggctc tattgaacct acagatctac tgctgcaaaa
     2341 ccccatcact gtccagcaag gcttctgcag agactccagg ctccgagtcc aagggcaaag
     2401 cccagcttct ctattacgtg aacttgctgt taatagacca ccgtttcctc ctccgccacg
     2461 gggactatgt gctccacatg tggcagatat ctggcaaggc agaggagcag ggcagcttca
     2521 atgctgacaa gctcacatcc gcaaccaatc ctgacaagga gaactcaatg tccatttcca
     2581 tcctgctgga caattactgt caccccatag ctttgcctaa gcaccggccc acccctgacc
     2641 cagagggaga cagggttcgg gctgaaatgc ccaatcagct tcgaaagcaa ttggaggcga
     2701 tcatagccac agatccactt aaccccctca cagcagagga caaagaattg ctctggcatt
     2761 ttcgatatga aagcctgaag catccgaagg cttaccctaa gctattcagc tcagtgaaat
     2821 gggggcagca agaaattgtt gccaaaacgt accagctgtt agccagaagg gagatctggg
     2881 atcaaagtgc tttggacgtt ggcttaacca tgcagctcct ggactgcaac ttttcagacg
     2941 agaatgtccg ggccattgca gttcagaaac tggagagctt agaggacgat gacgttttac
     3001 attaccttct ccagctggta caggctgtga aatttgaacc gtaccacgac agtgcgctgg
     3061 ccagattcct gctgaagcgt ggcttgagga acaaaagaat cggtcacttc ttgttctggt
     3121 tcctgcgaag tgagatcgca cagtccagac actatcagca gaggttcgct gtgatcctgg
     3181 aggcgtacct gcgaggctgt ggcacagcca tgttgcagga cttcacacag caggtccatg
     3241 tgattgagat gttacagaaa gtcaccattg atattaaatc gctctcggca gagaagtatg
     3301 acgtcagttc ccaagttatt tcacagctta agcaaaagct tgaaagcctt cagaactcca
     3361 atctccccga gagctttaga gttccctatg atcctggact aaaagccggt accctggtga
     3421 tcgagaaatg caaagtgatg gcctccaaga agaagcccct gtggcttgag tttaagtgtg
     3481 ctgatcccac agtcctatcc aacgaaacca ttggaatcat ctttaaacat ggtgatgatc
     3541 tgcgccaaga catgttgatc ttgcagattc tacgcatcat ggagtccatt tgggagactg
     3601 aatctctgga cctgtgcctt ctgccttacg gttgcatctc aactggtgac aaaataggaa
     3661 tgatcgagat tgtaaaggat gccacaacga tcgctcaaat tcagcaaagc acagtgggta
     3721 acacgggggc attcaaagat gaagtcctga atcactggct caaggaaaaa tgtcctattg
     3781 aagaaaagtt tcaggccgca gtggaaaggt ttgtttactc ctgtgcaggc tactgtgtgg
     3841 ccacatttgt tcttgggatc ggtgacaggc acaacgacaa cattatgatc tcagagacag
     3901 gaaacctatt tcatatagac ttcggacaca ttcttgggaa ttacaagagt ttcctgggca
     3961 tcaataaaga gagagtgccc ttcgtcctaa ccccagactt cttgtttgtg atgggatctt
     4021 ctggaaaaaa gacaagtcca cacttccaga aattccagga tgtctgtgtt agagcttacc
     4081 tagctcttcg ccatcacaca aacctgttga tcatcttgtt ctccatgatg ctgatgacag
     4141 gaatgcccca gctgacaagc aaagaggaca ttgaatatat ccgggatgcc ctcaccgtgg
     4201 gaaaaagcga ggaggacgct aagaaatatt tccttgatca gatcgaagtc tgcagagaca
     4261 aaggatggac tgtgcagttt aactggttcc tacatcttgt tcttggcatc aaacaaggag
     4321 aaaagcactc cgctacgcgt acgcggccgc tcgagcagaa actcatctca gaagaggatc
     4381 tggcagcaaa tgatatcctg gattacaagg atgacgacga taaggtttaa acggccggcc
     4441 gcggtcatag ctgtttcctg aacagatccc gggtggcatc cctgtgaccc ctccccagtg
     4501 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     4561 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     4621 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     4681 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     4741 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     4801 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     4861 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     4921 tgaaccactg ctcccttccc tgtccttctg attttaaaat aactatacca gcaggaggac
     4981 gtccagacac agcataggct acctggccat gcccaaccgg tgggacattt gagttgcttg
     5041 cttggcactg tcctctcatg cgttgggtcc actcagtaga tgcctgttga attgggtacg
     5101 cggccagcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5161 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5221 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5281 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5341 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5401 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5461 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5521 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5581 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5641 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5701 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5761 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5821 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5881 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5941 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg aggctatggc
     6001 agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg
     6061 gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc
     6121 gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc caacaccgtg
     6181 cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt
     6241 ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac tccagcatga gatccccgcg
     6301 ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa
     6361 ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc
     6421 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     6481 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
     6541 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcgatccgc cacacccagc
     6601 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     6661 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg
     6721 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
     6781 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     6841 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     6901 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     6961 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
     7021 gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg
     7081 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     7141 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     7201 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga
     7261 tcgatctttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     7321 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     7381 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     7441 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     7501 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     7561 ggagcctggg gactttccac accctaactg acacacattc cacagctggt tctttccgcc
     7621 tcaggactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7681 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7741 cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     7801 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     7861 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     7921 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     7981 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     8041 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     8101 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     8161 atttaacaaa aatttaacgc gaatttt
//
