LOCUS V019391 7697 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019391 VERSION V019391 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 671..2377 /label="PRKAG2(NM_016203)" /note="PRKAG2(NM_016203)" /gene="PRKAG2" CDS 2384..2449 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2473..3061 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3127..3175 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3377..3805 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3819..4148 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4215..5009 /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..6345) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6516..7376) /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(7377..7481) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg 661 taccgccacc atgggaagcg cggttatgga caccaagaag aaaaaagatg tttccagccc 721 cggcgggagc ggcggcaaga aaaatgccag ccagaagagg cgttcgctgc gcgtgcacat 781 tccggacctg agctccttcg ccatgccgct cctggacgga gacctggagg gttccggaaa 841 gcattcctct cgaaaggtgg acagcccctt cggcccgggc agcccctcca aagggttctt 901 ctccagaggc ccccagcccc ggccctccag ccccatgtct gcacctgtga ggcccaagac 961 cagccccggc tctcccaaaa ccgtgttccc gttctcctac caggagtccc cgccacgctc 1021 ccctcgacgc atgagcttca gtgggatctt ccgctcctcc tccaaagagt cttcccccaa 1081 ctccaaccct gctacctcgc ccgggggcat caggtttttc tcccgctcca gaaaaacctc 1141 cggcctctcc tcctctccgt caacacccac ccaagtgacc aagcagcaca cgtttcccct 1201 ggaatcctat aagcacgagc ctgaacggtt agagaatcgc atctatgcct cgtcttcccc 1261 cccggacaca gggcagaggt tctgcccgtc ttccttccag agcccgacca ggcctccact 1321 ggcatcaccg acacactatg ctccctccaa agccgcggcg ctggcggcgg ccctgggacc 1381 cgcggaagcc ggcatgctgg agaagctgga gttcgaggac gaagcagtag aagactcaga 1441 aagtggtgtt tacatgcgat tcatgaggtc acacaagtgt tatgacatcg ttccaaccag 1501 ttcaaagctt gttgtctttg atactacatt acaagttaaa aaggccttct ttgctttggt 1561 agccaacggt gtccgagcag cgccactgtg ggagagtaaa aaacaaagtt ttgtaggaat 1621 gctaacaatt acagatttca taaatatact acatagatac tataaatcac ctatggtaca 1681 gatttatgaa ttagaggaac ataaaattga aacatggagg gagctttatt tacaagaaac 1741 atttaagcct ttagtgaata tatctccaga tgcaagcctc ttcgatgctg tatactcctt 1801 gatcaaaaat aaaatccaca gattgcccgt tattgaccct atcagtggga atgcacttta 1861 tatacttacc cacaaaagaa tcctcaagtt cctccagctt tttatgtctg atatgccaaa 1921 gcctgccttc atgaagcaga acctggatga gcttggaata ggaacgtacc acaacattgc 1981 cttcatacat ccagacactc ccatcatcaa agccttgaac atatttgtgg aaagacgaat 2041 atcagctctg cctgttgtgg atgagtcagg aaaagttgta gatatttatt ccaaatttga 2101 tgtaattaat cttgctgctg agaaaacata caataaccta gatatcacgg tgacccaggc 2161 ccttcagcac cgttcacagt attttgaagg tgttgtgaag tgcaataagc tggaaatact 2221 ggagaccatc gtggacagaa tagtaagagc tgaggtccat cggctggtgg tggtaaatga 2281 agcagatagt attgtgggta ttatttccct gtcggacatt ctgcaagccc tgatcctcac 2341 accagcaggt gccaaacaaa aggagacaga aacggagctc gaggactaca aagaccatga 2401 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc 2461 cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt 2521 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct 2581 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt 2641 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac 2701 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct 2761 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca 2821 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt 2881 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc 2941 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct 3001 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg 3061 cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg 3121 ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa 3181 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc 3241 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca 3301 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc 3361 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 3421 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 3481 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct 3541 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 3601 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 3661 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc 3721 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg 3781 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa 3841 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 3901 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 3961 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 4021 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 4081 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 4141 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat 4201 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg 4261 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg 4321 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg 4381 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc 4441 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg 4501 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca 4561 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc 4621 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg 4681 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg 4741 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata 4801 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg 4861 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat 4921 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct 4981 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga 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 aatgctcacg 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 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc 6241 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 6301 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 6361 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 6421 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 6481 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 6541 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 6601 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 6661 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 6721 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 6781 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 6841 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 6901 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 6961 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 7021 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 7081 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 7141 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 7201 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 7261 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 7321 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 7381 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 7441 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 7501 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa 7561 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg 7621 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca 7681 tgaagaatct gcttagg //