LOCUS V025569 6722 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025569 VERSION V025569 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" CDS 941..2212 /label="Acat1(NM_144784)" /note="Acat1(NM_144784)" /gene="Acat1" CDS 2219..2284 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2322..2546 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2592..3020 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3034..3363 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3430..4224 /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 4398..4519 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4568..4584) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4592..4608 /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(4616..4646) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4661..4682 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4970..5555) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5726..6586) /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(6587..6691) /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 aagcttggta ccgagctcgg atccgccacc atggctgccc tggtggctct 961 gcacggcgtc gtccgcaggc ctctgcttcg cgggctgctg caggaagtaa gatgcctgga 1021 acgaagttat gcatccaaac ccactttgaa tgaagtggtt atagtaagtg ctataagaac 1081 tcccattgga tccttcctgg gcagccttgc ctctcagccg gccactaaac ttggtactgc 1141 tgcaattcag ggagccattg agaaggcagg gattccaaaa gaagaagtga aggaagtcta 1201 catgggcaat gtcatccaag ggggtgaagg acaggcccct accaggcaag caacactggg 1261 cgcaggttta cctatttcca ctccatgcac cacagtaaac aaggtttgtg cttcaggaat 1321 gaaagccatc atgatggcct ctcaaagtct tatgtgtgga catcaggatg tgatggtggc 1381 aggcgggatg gagagcatgt ccaatgtccc atacgtaatg agcagaggag caacaccata 1441 tggtggggta aaacttgaag acctgattgt aaaagacggg ctaactgatg tctacaataa 1501 aattcatatg ggtaactgtg ctgagaatac tgcaaagaag atgaatatct cacggcagga 1561 acaggatacg tacgctctca gctcttacac cagaagtaaa gaagcgtggg acgcagggaa 1621 gtttgccagt gagattactc ccatcaccat ctcagtgaaa ggtaaaccag atgtggtggt 1681 gaaagaagat gaagaataca agcgtgttga ctttagtaaa gtgccaaagc tcaagaccgt 1741 gttccagaaa gaaaatggca caataacagc tgccaatgcc agcacactga acgatggagc 1801 agctgctctg gttctcatga ctgcagaggc agcccagagg ctcaatgtta agccattggc 1861 acgaattgca gcatttgctg atgctgccgt agaccccatt gattttccac ttgcgcctgc 1921 atatgccgta cctaaggttc ttaaatatgc aggactgaaa aaagaagaca ttgccatgtg 1981 ggaagtaaat gaagcattca gtgtggttgt gctagccaac attaaaatgc tggagattga 2041 cccccaaaaa gtaaatatcc acggaggagc tgtttctctg ggccatccaa ttgggatgtc 2101 tggagcccgg attgttgttc atatggctca tgccctgaag ccaggagagt tcggtctggc 2161 tagtatttgc aacggaggag gaggtgcttc cgccctgctg attgagaagc tgctcgagga 2221 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga 2281 caagtagtga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca 2341 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac 2401 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat 2461 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca 2521 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag 2581 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 2641 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 2701 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 2761 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 2821 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 2881 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 2941 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 3001 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc 3061 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg 3121 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 3181 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 3241 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 3301 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc 3361 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga 3421 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag 3481 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc 3541 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg 3601 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc 3661 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg 3721 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct 3781 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg 3841 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat 3901 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc 3961 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg 4021 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc 4081 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct 4141 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat 4201 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga 4261 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct 4321 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg 4381 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata 4441 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 4501 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt 4561 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 4621 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 4681 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 4741 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 4801 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 4861 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 4921 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 4981 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 5041 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 5101 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 5161 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 5221 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 5281 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 5341 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 5401 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 5461 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca 5521 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 5581 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 5641 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 5701 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 5761 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 5821 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 5881 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 5941 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 6001 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 6061 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 6121 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 6181 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 6241 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 6301 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 6361 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 6421 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 6481 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 6541 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 6601 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 6661 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 6721 tc //