LOCUS V045259 8062 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045259 VERSION V045259 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 925..3813 /label="USP4(NM_003363)" /note="USP4(NM_003363)" /gene="USP4" CDS 3826..3891 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3926..4150 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4196..4624 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4638..4967 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4979..5578 /codon_start=1 /gene="pac from Streptomyces alboniger" /note="confers resistance to puromycin" /product="puromycin N-acetyltransferase" /transl_table=1 /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*" /label="PuroR" polyA_signal 5738..5859 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5908..5924) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5932..5948 /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(5956..5986) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6001..6022 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6310..6895) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7066..7926) /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(7927..8031) /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 aagctttcta gagaattcgc caccatggcg gaaggtggag gctgccgtga gcgaccggat 961 gcggagactc agaagtccga gcttggaccc ttaatgagga ccacactcca acgcggggcg 1021 cagtggtatc ttattgacag ccggtggttc aagcagtgga agaagtatgt gggctttgac 1081 agctgggaca tgtacaatgt gggtgaacat aacctatttc ctggcccaat agacaactct 1141 gggctatttt cagatcctga gagtcagacc ttgaaagaac acttaattga tgaattggac 1201 tatgtattgg tccctaccga ggcgtggaat aaactactaa actggtacgg ctgtgtagaa 1261 ggccagcaac ccatcgtcag aaaagttgtg gagcatggcc tgtttgtcaa gcactgcaaa 1321 gtcgaggtgt atttgctgga actgaagctc tgtgagaaca gtgaccccac caatgtgctg 1381 agttgccatt tcagcaaggc agacaccatt gcaaccatcg agaaagagat gcggaagcta 1441 ttcaacatcc ctgcggagcg tgaaacacgg ctctggaaca aatacatgag caacacctac 1501 gagcagttga gcaagctaga caacactgtc caggatgctg ggctatacca gggtcaggtg 1561 ctagtaattg agcctcaaaa tgaagatggc acatggccca ggcagacctt gcagtcaaaa 1621 tcaagcactg cgcctagcag aaattttact acctctccaa aatcatcagc aagtccctat 1681 tcctcagtgt ctgcctctct cattgcaaat ggtgatagca ctagcacctg tgggatgcac 1741 agttccggtg tcagcagggg tggatctggc ttttctgctt cgtataattg tcaggagcca 1801 ccatcctctc atatacaacc tgggctctgt ggacttggaa acctgggaaa cacctgcttc 1861 atgaactccg ctttgcagtg tttgagcaac actgcaccac tgactgacta ctttctcaaa 1921 gatgagtatg aagccgaaat caacagagac aaccctctgg ggatgaaagg ggaaattgca 1981 gaagcctatg ctgaactcat taagcagatg tggtctggaa gggacgccca tgtggcacct 2041 cgcatgttca aaactcaagt aggacgtttt gctcctcaat tttctggcta ccagcaacaa 2101 gattctcagg agctgctggc ctttcttcta gatggattgc atgaagatct gaaccgggta 2161 aagaaaaagc cctacttgga gctgaaggat gccaatgggc ggccagatgc ggtggtggca 2221 aaggaagcct gggagaatca caggttgagg aatgattctg tgattgtgga tactttccat 2281 ggcctcttca aatctacttt ggtttgccca gaatgtgcta aggtttctgt gacctttgac 2341 ccattttgct atctaacgct gccactgccc ttgaagaaag atcgagttat ggaggttttc 2401 ctggttcctg ctgaccctca ctgcagacct actcagtacc gtgtgactgt gccgctgatg 2461 ggggctgtgt ccgacctgtg cgaggctctc tccaggctgt ctggcattgc tgcagaaaat 2521 atggtggtcg cagatgtgta taatcaccga ttccacaaaa ttttccaaat ggatgaaggt 2581 ttaaaccaca tcatgcctcg ggatgacatt ttcgtgtacg aggtctgcag cacttccgtg 2641 gatggctcgg aatgtgtcac gcttccagtc tacttcaggg agaggaagtc caggccatca 2701 agcacttcct ccgcatcagc gctatatggg cagccactat tgctttctgt ccccaagcac 2761 aagttaaccc ttgagtcttt gtaccaggct gtttgtgatc gtatcagccg ctatgtgaaa 2821 cagcctttac ctgatgagtt tggcagctca cccttggagc caggggcctg caatggctcc 2881 aggaacagct gtgaaggaga agatgaggaa gaaatggagc atcaggaaga aggcaaagag 2941 cagctttcag aaacagaagg cagtggggaa gatgagccag gaaatgaccc cagtgagacc 3001 acccaaaaga agatcaaagg ccagccctgc ccaaaaaggc tttttacctt cagtcttgtg 3061 aactcctatg gaacagctga cataaattca cttgcagctg atggaaaact acttaaactc 3121 aactctcgat ctacactggc catggattgg gacagtgaaa ctcggagact ttactatgat 3181 gagcaagaat ctgaggccta cgagaagcat gtgagcatgt tgcagcctca gaagaagaag 3241 aagaccacag tggccctgag agactgcatc gagctcttca ccaccatgga gacccttggg 3301 gagcatgacc cctggtactg tcccaactgt aagaagcatc aacaggccac aaaaaagttt 3361 gacctatggt ccttgcccaa gatcctggtg gtccacctca aacgtttctc ctacaacaga 3421 tactggaggg ataagctcga cacagtcgta gaattcccaa tcagagggct gaacatgtcc 3481 gagtttgtct gtaacctgtc agcaaggcct tatgtgtacg acctcattgc cgtgtccaat 3541 cattatggag ccatgggggt tggccactac actgcatatg cgaagaacaa actgaatggt 3601 aaatggtatt actttgatga tagcaacgtg tccctggcct ctgaggatca gatagtgact 3661 aaagcagctt atgtgctatt ttaccaacgt cgagatgatg aattttataa gacaccttca 3721 cttagcagtt ctggttcctc tgatggaggg acacgaccaa gcagctctca gcagggcttt 3781 ggggatgatg aggcttgcag catggacacc aacctcgaga ccggtgacta caaagaccat 3841 gacggtgatt ataaagatca tgacatcgat tacaaggatg acgatgacaa gtgaccggta 3901 agtttaaacc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc 3961 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa 4021 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg 4081 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg 4141 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg 4201 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 4261 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 4321 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 4381 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 4441 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 4501 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 4561 attttggcca tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 4621 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag 4681 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 4741 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 4801 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 4861 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat 4921 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggat 4981 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg 5041 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg 5101 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat 5161 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag 5221 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc 5281 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc 5341 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag 5401 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga 5461 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga 5521 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgatt 5581 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc 5641 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca 5701 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa 5761 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 5821 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac 5881 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 5941 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 6001 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 6061 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 6121 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 6181 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 6241 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 6301 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 6361 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 6421 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 6481 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 6541 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 6601 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 6661 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 6721 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa 6781 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 6841 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 6901 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 6961 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 7021 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 7081 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 7141 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 7201 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 7261 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 7321 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 7381 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 7441 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 7501 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 7561 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 7621 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 7681 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 7741 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 7801 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 7861 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 7921 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 7981 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 8041 ccgaaaagtg ccacctgacg tc //