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 //