LOCUS V020834 8996 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020834 VERSION V020834 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..3676 /label="MAMLD1(NM_001177465)" /note="MAMLD1(NM_001177465)" /gene="MAMLD1" CDS 3683..3748 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 3772..4360 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 4426..4474 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 4676..5104 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 5118..5447 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5514..6308 /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 6484..6605 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6654..6670) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6678..6694 /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(6702..6732) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6747..6768 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7056..7644) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7815..8675) /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(8676..8780) /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 ccatggatga ctggaaaagt cggcttgtaa tcaagagcat 721 gcttccccat ttcgccatgg tgggaaatcg tcaggagccc agaaagctcc aggaatcggg 781 aactgttaag aggagacaag aagaagacca cttccagttt ccagacatgg ctgatggggg 841 ctaccctaat aaaattaaga ggccttgcct tgaagatgtc acccttgcaa tgggcccagg 901 tgctcatcct agtactgctt gtgcagaact gcaggtccct ccattgacaa taaatcctag 961 ccctgcggct atgggagtgg ctggccagtc attactgctg gagaataacc ctatgaatgg 1021 caacatcatg ggctcaccat ttgtagtacc acagactaca gaagtgggac tgaaagggcc 1081 cactgttcct tactatgaga aaatcaacag cgtgccggct gtagaccagg agcttcaaga 1141 gctgctagag gagctcacca aaattcaaga cccttctcca aatgagctag atcttgagaa 1201 gatactgggg acgaagccag aagagccact ggttttagat catccccagg caaccctaag 1261 cacaactccc aagccttcgg ttcagatgtc acacttggag agcctggctt ccagcaagga 1321 gtttgcttct agttgcagcc aagttactgg catgtcactt cagatcccat cctcctccac 1381 agggatcagc tattcgattc cttccaccag taagcagata gtgtcaccga gttcttcaat 1441 ggcacagtcc aagagccagg tccaggccat gctccctgtc gctctgcccc ccttaccagt 1501 gcctcagtgg catcacgccc accagctgaa ggcgttggca gccagcaagc aggggtctgc 1561 tacaaagcag caagggccca cccccagttg gtctggtctg cctcctccag gactctctcc 1621 accttaccgc ccagtgccat caccacaccc accaccgctg ccactgccac caccaccacc 1681 cccattcagc ccccagagcc tcatggtgtc ctgcatgtcg tccaatacct tgtcgggtag 1741 cactctccga ggctctccca atgccttact gtcaagcatg acgtccagca gcaatgctgc 1801 cctggggccc gccatgccct atgctcctga gaagctcccc agccctgctc tcactcaaca 1861 gccgcagttc ggccctcaga gctccattct tgccaacctc atgtcctcta ccatcaaaac 1921 ccctcaagga cacctgatgt ctgctctgcc tgccagcaac cctgggccgt ccccacccta 1981 tcgcccagag aagctctcta gcccaggctt gccacagcag tccttcaccc cacagtgttc 2041 cctgatccga agcctcactc ccaccagtaa tcttctaagc cagcaacagc agcagcagca 2101 gcagcagcag caagcaaatg tgatctttaa gcccataagc agcaactcat ccaaaaccct 2161 gagcatgatc atgcagcagg ggatggcaag ctccagccca ggagccacgg agccatttac 2221 ttttggcaac accaagccct tgtcccattt tgtttctgag ccgggtcccc agaagatgcc 2281 ctccatgcct accacctcta ggcagccttc cctgctccac tacctgcagc agccgacacc 2341 aacgcaggcc tcctcagcca ctgcctcctc cacggccact gccaccttgc agctgcagca 2401 gcagcagcag caacagcagc agcagcctga ccattcttca ttccttctgc agcagatgat 2461 gcagcaaccc cagcgttttc agcgatcagt ggcctcagat tccatgcctg ctctgcccag 2521 acagcaagag gaacagagat caggtcttat ggcaatgacc cctgaacggc agaatgcata 2581 tatctcccaa cagatgagtc catttgaagc cgtccaagaa caagtcacct ccaagtgtag 2641 ccggatcaag gcaagccccc catctagcaa gcacttgatg ccacccagaa ctgggcttct 2701 tcagaacaat ctgagtccag gaatgatccc actcaccagg caccagagct gcgagggcat 2761 gggagtgatc tcaccaactc tggggaagcg gcaaggaatt ttcacctcca gcccccagtg 2821 tcccatcctc tcacactcag gccagactcc cctgggcaga cttgactctg tctgccagca 2881 tatgcagagt cccaaggcca ccccaccaga agtgcccctg cctgggttct gtcccagctc 2941 cctgggcacc cagtccttga gtccccacca gctcagacgg cctagtgtgc caagaatgcc 3001 cactgcgttc aacaatgctg catgggtcac agcggcagca gctgtgacca cagcagtttc 3061 ggggaaaaca cccctcagcc aagtggataa tagcgttcag cagcactcac cttctggcca 3121 ggcctgcctt cagaggccat ctgattggga ggcacaagtg cccgctgcga tgggaacaca 3181 agtgcccctg gccaacaacc ccagcttcag cctgctgggc agccagagcc tcaggcagag 3241 cccggtacag ggcccggtgc ctgtagcaaa caccaccaag ttcctccagc agggtatggc 3301 cagctttagt cccctgagcc ccatacaggg catcgagcca ccaagctatg tggctgctgc 3361 tgccaccgct gctgctgctt ctgccgttgc tgccagccag ttcccaggtc cgttcgacag 3421 aacggatatt ccccctgagc tgccacctgc cgactttttg cgccagcccc aacccccact 3481 aaatgatctg atttcgtcac ctgactgcaa tgaggtagat ttcattgaag ctctcttgaa 3541 aggctcctgt gtgagcccag atgaagactg ggtgtgcaac ttgaggctga tcgacgacat 3601 tttggaacag catgctgctg ctcaaaatgc cacagcccag aattctgggc aagtcaccca 3661 ggatgctggg gcacttctcg aggactacaa agaccatgac ggtgattata aagatcatga 3721 catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga caatcaacct 3781 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 3841 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 3901 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 3961 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 4021 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 4081 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 4141 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt 4201 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 4261 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 4321 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa 4381 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac 4441 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc 4501 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc 4561 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg 4621 gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg 4681 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 4741 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 4801 gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct 4861 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 4921 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 4981 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 5041 attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 5101 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag 5161 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 5221 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 5281 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 5341 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat 5401 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag 5461 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg 5521 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 5581 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 5641 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg 5701 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 5761 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 5821 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 5881 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 5941 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 6001 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg 6061 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 6121 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 6181 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 6241 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 6301 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca 6361 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc 6421 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac 6481 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 6541 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 6601 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag 6661 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 6721 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 6781 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 6841 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 6901 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 6961 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 7021 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 7081 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 7141 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 7201 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc 7261 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 7321 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 7381 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 7441 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca 7501 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 7561 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 7621 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 7681 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 7741 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 7801 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 7861 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 7921 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 7981 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 8041 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 8101 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 8161 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 8221 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 8281 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 8341 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 8401 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 8461 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 8521 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 8581 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 8641 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 8701 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 8761 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg 8821 ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt 8881 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa 8941 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg //