LOCUS       V024476                 6950 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024476
VERSION     V024476
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             928..1017
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     CDS             1033..2472
                     /label="Akt1(NM_001331107)"
                     /note="Akt1(NM_001331107)"
                     /gene="Akt1"
     polyA_signal    2550..2774
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2820..3248
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3262..3591
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3658..4452
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    4626..4747
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4796..4812)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4820..4836
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(4844..4874)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4889..4910
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5198..5783)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5954..6814)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(6815..6919)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagccaccat ggcggcggaa caaaaactaa tatcagagga agatttggaa
      961 cagaaactaa tcagtgaaga agatttagag caaaagttaa tttccgagga ggacttatca
     1021 gcggaaaagc ttatgaacga cgtagccatt gtgaaggagg gctggctgca caaacgaggg
     1081 gaatatatta aaacctggcg gccacgctac ttcctcctca agaacgatgg cacctttatt
     1141 ggctacaagg aacggcctca ggatgtggat cagcgagagt ccccactcaa caacttctca
     1201 gtggcacaat gccagctgat gaagacagag cggccaaggc ccaacacctt tatcatccgc
     1261 tgcctgcagt ggaccacagt cattgagcgc accttccatg tggaaacgcc tgaggagcgg
     1321 gaagaatggg ccaccgccat tcagactgtg gcagatggac tcaagaggca ggaagaagag
     1381 acgatggact tccgatcagg ctcacccagt gacaactcag gggctgaaga gatggaggtg
     1441 tccctggcca agcccaagca ccgtgtgacc atgaacgagt ttgagtacct gaagctactg
     1501 ggcaagggca cctttgggaa ggtgattctg gtgaaagaga aggccacagg ccgctactat
     1561 gccatgaaga tcctcaagaa ggaggtcatc gtcgccaagg atgaggttgc ccacacgctt
     1621 actgagaacc gtgtcctgca gaactctagg catcccttcc ttacggccct caagtactca
     1681 ttccagaccc acgaccgcct ctgctttgtc atggagtatg ccaacggggg cgagctcttc
     1741 ttccacctgt ctcgagagcg tgtgttctcc gaggaccggg cccgcttcta tggtgcggag
     1801 attgtgtctg ccctggacta cttgcactcc gagaagaacg tggtgtaccg ggacctgaag
     1861 ctggagaacc tcatgctgga caaggacggg cacatcaaga taacggactt cgggctgtgc
     1921 aaggagggga tcaaggacgg tgccactatg aagacattct gcggaacgcc ggagtacctg
     1981 gcccctgagg tgctggagga caacgactac ggccgtgcag tggactggtg ggggctgggc
     2041 gtggtcatgt acgagatgat gtgtggccgc ctgcccttct acaaccagga ccacgagaag
     2101 ctgttcgagc tgatcctcat ggaggagatc cgcttcccgc gcacactcgg ccctgaggcc
     2161 aagtccctgc tctccgggct gctcaagaag gaccctacac agaggctcgg tgggggctcc
     2221 gaggatgcca aggagatcat gcagcaccgg ttctttgcca acatcgtgtg gcaggatgtg
     2281 tatgagaaga agctgagccc acctttcaag ccccaggtca cctctgagac tgacaccagg
     2341 tatttcgatg aggagttcac agctcagatg atcaccatca cgccgcctga tcaagatgac
     2401 agcatggagt gtgtggacag tgagcggagg ccgcacttcc cccagttctc ctactcagcc
     2461 agtggcacag cctgaattct gcagatatcc agcacagtgg cggccgctcg agtctagagg
     2521 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     2581 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2641 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     2701 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     2761 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     2821 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2881 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2941 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3001 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3061 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3121 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3181 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3241 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3301 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3361 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3421 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3481 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3541 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     3601 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     3661 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3721 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3781 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3841 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3901 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3961 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4021 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4081 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4141 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4201 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4261 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4321 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4381 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4441 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4501 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4561 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4621 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4681 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4741 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     4801 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4861 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4921 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4981 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5041 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5101 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5161 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5221 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5281 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5341 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5401 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5461 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5521 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5581 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5641 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5701 cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta
     5761 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5821 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5881 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5941 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6001 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6061 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6121 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6181 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6241 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6301 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6361 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6421 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6481 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6541 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6601 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6661 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6721 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6781 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6841 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6901 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
