LOCUS V020250 6230 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020250 VERSION V020250 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..910 /label="Snrpg(NM_026506)" /note="Snrpg(NM_026506)" /gene="Snrpg" CDS 917..982 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1006..1594 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 1660..1708 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 1910..2338 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2352..2681 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2748..3542 /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 3718..3839 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3888..3904) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3912..3928 /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(3936..3966) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3981..4002 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4290..4878) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5049..5909) /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(5910..6014) /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 ccatgagcaa agcccaccct cccgagctga agaagtttat 721 ggacaagaag ttatcattga agttaaacgg tggcaggcat gtccaaggaa tactgcgggg 781 ctttgatccc tttatgaacc ttgtgattga cgagtgtgtg gagatggcaa ccagtgggca 841 acagaacaac attggcatgg tggtcatccg aggaaacagc atcatcatgt tagaagcctt 901 ggaaagagtc ctcgaggact acaaagacca tgacggtgat tataaagatc atgacatcga 961 ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca acctctggat 1021 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 1081 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 1141 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 1201 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 1261 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 1321 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 1381 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc 1441 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 1501 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 1561 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa 1621 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata 1681 aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac 1741 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc 1801 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca 1861 ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt 1921 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc 1981 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc 2041 tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg 2101 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga 2161 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc 2221 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg 2281 gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa 2341 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa 2401 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc 2461 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc 2521 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct 2581 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga 2641 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta 2701 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag 2761 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg 2821 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc 2881 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag 2941 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca 3001 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat 3061 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata 3121 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac 3181 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc 3241 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg 3301 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg 3361 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta 3421 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg 3481 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct 3541 gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga 3601 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac 3661 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac 3721 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 3781 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 3841 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt 3901 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 3961 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 4021 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 4081 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 4141 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 4201 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 4261 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 4321 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 4381 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 4441 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg 4501 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 4561 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 4621 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 4681 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt 4741 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 4801 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 4861 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 4921 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 4981 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 5041 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 5101 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 5161 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 5221 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 5281 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 5341 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 5401 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 5461 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 5521 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 5581 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 5641 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 5701 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 5761 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 5821 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 5881 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 5941 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 6001 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat 6061 ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca tagttaagcc 6121 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag 6181 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg //