LOCUS       V032781                 7542 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032781
VERSION     V032781
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             872..2311
                     /label="Akt1(NM_033230)"
                     /note="Akt1(NM_033230)"
                     /gene="Akt1"
     CDS             2348..3058
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK*"
                     /label="mCherry"
     polyA_signal    3109..3333
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3379..3807
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3821..4150
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4217..5011
                     /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    5185..5306
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5355..5371)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5379..5395
                     /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(5403..5433)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5448..5469
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5757..6342)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6513..7373)
                     /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(7374..7478)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagccac catgaacgac gtagccattg tgaaggaggg
      901 ctggctgcac aaacgagggg aatatattaa aacctggcgg ccacgctact tcctcctcaa
      961 gaatgatggc acctttattg gctacaagga acggcctcag gatgtggagc agcgcgagtc
     1021 cccactcaac aacttctcag tggcacaatg tcagctgatg aagacagagc ggccgaggcc
     1081 caacaccttc atcatccgct gcctgcagtg gaccacggtc attgagcgca ccttccatgt
     1141 ggaaacgcct gaggagcggg aagagtggac caccgccatt cagactgtgg ctgatggact
     1201 caaacggcag gaggaggaga cgatggactt ccggtcaggt tcacccagtg acaactcagg
     1261 tgctgaggag atggaggtgg ccctggccaa gcccaagcac cgtgtgacca tgaacgagtt
     1321 tgagtacctg aagctactgg gcaagggcac ctttgggaag gtgatcctgg tgaaggagaa
     1381 ggccacaggt cgctactatg ccatgaagat cctcaagaag gaggtcatcg ttgccaagga
     1441 tgaggttgcc cacacgctta ctgagaaccg tgtcctgcag aactctaggc atcccttcct
     1501 tacagccctc aagtactcat tccagaccca cgaccgcctc tgctttgtca tggagtacgc
     1561 caatgggggc gagctcttct tccacctgtc tcgtgagcgc gtgttttcag aggaccgggc
     1621 ccgcttctac ggtgcggaga ttgtgtccgc cctggactac ttgcactccg agaagaacgt
     1681 ggtgtaccgg gacctcaagc tggagaacct catgctggac aaggacgggc acatcaagat
     1741 aacggacttc gggctgtgca aggagggtat caaggacggt gccaccatga agacgttctg
     1801 cgggacaccc gagtacctgg cccctgaggt gctggaggac aacgactatg gccgtgcagt
     1861 ggactggtgg gggctgggcg tggtcatgta cgagatgatg tgcggccgcc tgcccttcta
     1921 caaccaggac catgagaagc tgttcgagct catcctaatg gaggagatcc gcttcccacg
     1981 cacactcggg ccggaggcca agtccctgct ctcggggctg ctcaagaagg accctacaca
     2041 gaggctcggt gggggctccg aggacgccaa ggagatcatg cagcaccgct tctttgccaa
     2101 catcgtgtgg caagatgtgt atgagaagaa gctgagccca cctttcaagc cccaggtcac
     2161 ctctgagacc gacaccaggt attttgatga ggagttcaca gctcagatga tcaccatcac
     2221 gccgcctgat caagatgaca gcatggagtg tgtggacagt gaacggaggc cgcactttcc
     2281 ccagttctcc tactcagcca gtggcacagc cgaattcggt ggaggcggat caggaggcgg
     2341 tggatctatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
     2401 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     2461 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     2521 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     2581 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     2641 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     2701 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     2761 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     2821 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     2881 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     2941 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     3001 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaggc
     3061 ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
     3121 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     3181 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     3241 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     3301 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     3361 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3421 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3481 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3541 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3601 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3661 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3721 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3781 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     3841 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3901 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3961 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4021 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4081 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4141 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     4201 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4261 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4321 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4381 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4441 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4501 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4561 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4621 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4681 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4741 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4801 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4861 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4921 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4981 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     5041 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5101 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5161 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     5221 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5281 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5341 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5401 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5461 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5521 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5581 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5641 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5701 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5761 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5821 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5881 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5941 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6001 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6061 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6121 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6181 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6241 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt
     6301 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6361 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6421 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6481 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6541 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6601 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6661 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6721 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6781 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6841 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6901 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6961 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7021 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7081 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7141 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7201 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7261 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7321 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7381 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7441 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7501 cctgacgtcg acggatcggg agatctcccg atcccctatg gt
//
