LOCUS V019328 6629 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019328 VERSION V019328 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..1309 /label="FAIM(NM_001033030)" /note="FAIM(NM_001033030)" /gene="FAIM" CDS 1316..1381 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1405..1993 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2059..2107 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2309..2737 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2751..3080 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3147..3941 /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 4117..4238 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4287..4303) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4311..4327 /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(4335..4365) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4380..4401 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4689..5277) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5448..6308) /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(6309..6413) /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 atgctgcttc cctttataag gacactccca ctgttgtgct ataatcatct 721 cttggtatct ccggactctg ccactctgag ccctccttac agcctagaaa aaatgacaga 781 tctcgtagct gtttgggatg ttgctttaag tgacggagtc cacaagatcg aatttgaaca 841 tgggactaca tcaggcaaac gagtagtata tgtagatgga aaggaagaga taagaaaaga 901 gtggatgttc aaattagtgg gcaaagaaac attctatgtt ggagctgcaa agacaaaagc 961 gaccataaat atagacgcta tcagtggttt tgcttatgaa tatactctgg aaattaatgg 1021 gaaaagtctc aagaagtata tggaggacag atcaaaaacc accaatactt gggtattaca 1081 catggatggt gagaacttta gaattgtttt ggaaaaagat gctatggacg tatggtgcaa 1141 tggtaaaaaa ttggagacag cgggtgagtt tgtagatgat gggactgaaa ctcacttcag 1201 tatcgggaac catgactgtt acataaaggc tgtcagtagt gggaagcgga aagaagggat 1261 tattcatact ctcattgtgg ataatagaga aatcccagag attgcaagtc tcgaggacta 1321 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa 1381 gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg 1441 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 1501 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 1561 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 1621 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 1681 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 1741 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag 1801 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 1861 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 1921 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 1981 gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg 2041 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg 2101 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga 2161 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg 2221 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg 2281 cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg 2341 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct 2401 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 2461 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 2521 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg 2581 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac 2641 cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta 2701 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt 2761 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 2821 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 2881 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 2941 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 3001 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg 3061 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc 3121 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc 3181 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 3241 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 3301 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca 3361 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 3421 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 3481 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 3541 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 3601 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 3661 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 3721 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 3781 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 3841 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 3901 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc 3961 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc 4021 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag 4081 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat 4141 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 4201 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc 4261 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg 4321 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa 4381 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 4441 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt 4501 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga 4561 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca 4621 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 4681 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 4741 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 4801 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 4861 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc 4921 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 4981 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 5041 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 5101 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag 5161 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 5221 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 5281 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 5341 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 5401 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 5461 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 5521 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 5581 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 5641 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 5701 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 5761 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc 5821 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc 5881 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca 5941 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag 6001 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg 6061 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa 6121 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 6181 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga 6241 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga 6301 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 6361 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 6421 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga 6481 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg 6541 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga 6601 ccgacaattg catgaagaat ctgcttagg //