LOCUS V021331 7400 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021331 VERSION V021331 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 683..2047 /label="TNFRSF1A(NM_001065)" /note="TNFRSF1A(NM_001065)" /gene="TNFRSF1A" CDS 2087..2152 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2176..2764 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2830..2878 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3080..3508 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3522..3851 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3918..4712 /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 4888..5009 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5058..5074) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5082..5098 /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(5106..5136) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5151..5172 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5460..6048) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6219..7079) /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(7080..7184) /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 taccgagctc ggatccgcca ccatgggcct ctccaccgtg cctgacctgc tgctgccact 721 ggtgctcctg gagctgttgg tgggaatata cccctcaggg gttattggac tggtccctca 781 cctaggggac agggagaaga gagatagtgt gtgtccccaa ggaaaatata tccaccctca 841 aaataattcg atttgctgta ccaagtgcca caaaggaacc tacttgtaca atgactgtcc 901 aggcccgggg caggatacgg actgcaggga gtgtgagagc ggctccttca ccgcttcaga 961 aaaccacctc agacactgcc tcagctgctc caaatgccga aaggaaatgg gtcaggtgga 1021 gatctcttct tgcacagtgg accgggacac cgtgtgtggc tgcaggaaga accagtaccg 1081 gcattattgg agtgaaaacc ttttccagtg cttcaattgc agcctctgcc tcaatgggac 1141 cgtgcacctc tcctgccagg agaaacagaa caccgtgtgc acctgccatg caggtttctt 1201 tctaagagaa aacgagtgtg tctcctgtag taactgtaag aaaagcctgg agtgcacgaa 1261 gttgtgccta ccccagattg agaatgttaa gggcactgag gactcaggca ccacagtgct 1321 gttgcccctg gtcattttct ttggtctttg ccttttatcc ctcctcttca ttggtttaat 1381 gtatcgctac caacggtgga agtccaagct ctactccatt gtttgtggga aatcgacacc 1441 tgaaaaagag ggggagcttg aaggaactac tactaagccc ctggccccaa acccaagctt 1501 cagtcccact ccaggcttca cccccaccct gggcttcagt cccgtgccca gttccacctt 1561 cacctccagc tccacctata cccccggtga ctgtcccaac tttgcggctc cccgcagaga 1621 ggtggcacca ccctatcagg gggctgaccc catccttgcg acagccctcg cctccgaccc 1681 catccccaac ccccttcaga agtgggagga cagcgcccac aagccacaga gcctagacac 1741 tgatgacccc gcgacgctgt acgccgtggt ggagaacgtg cccccgttgc gctggaagga 1801 attcgtgcgg cgcctagggc tgagcgacca cgagatcgat cggctggagc tgcagaacgg 1861 gcgctgcctg cgcgaggcgc aatacagcat gctggcgacc tggaggcggc gcacgccgcg 1921 gcgcgaggcc acgctggagc tgctgggacg cgtgctccgc gacatggacc tgctgggctg 1981 cctggaggac atcgaggagg cgctttgcgg ccccgccgcc ctcccgcccg cgcccagtct 2041 tctcagagaa ttctgcagat atccagcaca gtggcggccg ctcgaggact acaaagacca 2101 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtgagg 2161 gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt 2221 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat 2281 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct 2341 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct 2401 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc 2461 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg 2521 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc 2581 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac 2641 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg 2701 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc 2761 ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc 2821 gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat 2881 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg 2941 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc 3001 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg 3061 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg 3121 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc 3181 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc 3241 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt 3301 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag 3361 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg 3421 gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag 3481 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg 3541 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 3601 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 3661 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc 3721 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg 3781 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag 3841 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag 3901 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt 3961 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg 4021 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg 4081 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg 4141 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg 4201 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca 4261 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc 4321 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc 4381 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca 4441 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga 4501 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg 4561 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg 4621 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg 4681 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc 4741 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa 4801 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat 4861 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa 4921 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt 4981 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag 5041 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 5101 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 5161 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 5221 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 5281 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 5341 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 5401 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 5461 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 5521 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 5581 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 5641 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 5701 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 5761 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 5821 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 5881 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 5941 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 6001 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 6061 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 6121 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 6181 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 6241 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 6301 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 6361 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 6421 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 6481 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 6541 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 6601 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 6661 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 6721 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 6781 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 6841 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 6901 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 6961 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 7021 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 7081 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 7141 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 7201 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta 7261 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg 7321 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt 7381 gcatgaagaa tctgcttagg //