LOCUS V044273 7474 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044273 VERSION V044273 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 706..2130 /label="Tfeb(NM_001161723)" /note="Tfeb(NM_001161723)" /gene="Tfeb" CDS 2137..2226 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" misc_feature 2250..2838 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2904..2952 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3154..3582 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3596..3925 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3992..4786 /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 4962..5083 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5132..5148) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5156..5172 /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(5180..5210) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5225..5246 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5534..6122) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6293..7153) /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(7154..7258) /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 ggatccacta gtccagtgtg gtggaattcg ccaccatggc gtcacgcatc 721 gggctgcgca tgcagctcat gcgggagcag gcccagcagg aggagcagcg agagcgcatg 781 cagcagcagg ctgtcatgca ttatatgcaa cagcagcagc agcagcagca gcagctgggt 841 gggcccccca ccccagccat caacacccct gtccacttcc agtcgccccc gcctgtgccc 901 ggggaggtgc tgaaggtgca gtcctacctg gagaacccca cctcctacca cctgcaacag 961 tcccagcatc agaaggttcg ggagtatctg tctgagacct atgggaacaa gtttgctgcc 1021 cacgtgagcc cagcccaagg ttccccgaag cctgccccag cagcatcccc aggggtgcgg 1081 gctggacacg tactgtccac ctcggccggc aacagtgctc ccaacagtcc catggccatg 1141 ctacatatca gctccaaccc cgagaaagag tttgatgatg tcattgacaa cattatgcgc 1201 ctggacagcg tgctgggcta catcaaccct gagatgcaga tgcctaacac gctgcccctg 1261 tctagcagcc acctgaacgt gtacagcggt gacccccagg tcacagcctc catggtgggt 1321 gtcaccagca gctcctgccc tgccgacctg actcagaagc gagagctaac agatgctgag 1381 agcagagccc tggccaagga gcggcagaag aaagacaatc acaacctaat tgagagaaga 1441 cgcaggttca acatcaatga ccggatcaag gagctgggaa tgctgatccc caaggccaac 1501 gacctggacg tgcgctggaa caaaggcacc atcctcaagg cctctgtgga ttacatccgg 1561 aggatgcaga aggacctgca gaagtcccgg gagctggaga accactcccg gcgcctggag 1621 atgactaaca agcagctctg gctccgcatc caggagctgg agatgcaggc acgcgtgcac 1681 ggcctcccca ccacctcgcc gtcgggtgtg aatatggccg agctggccca gcaggtggtg 1741 aagcaagagt tgcccagtga ggatggccca ggggaggcgc tgatgctggg gcctgaggtc 1801 cctgagcctg agcaaatgcc ggctcttcct ccccaggctc cgctgccctc ggccgcccag 1861 ccacagtctc cgttccatca cctggacttc agccatggcc tgagctttgg gggtgggggc 1921 gacgaggggc ccacaggtta ccccgatacc ctggggacag agcacggctc cccattcccc 1981 aacctgtcca agaaggatct ggacttaatg ctcctagatg actccctgct ccccctggcc 2041 tctgaccccc tcttttctac catgtctcct gaggcctcca aggccagcag ccgccggagc 2101 agcttcagca tggaggaggg tgatgttctg ctcgaggaac aaaaactaat atcagaggaa 2161 gatttggaac agaaactaat cagtgaagaa gatttagagc aaaagttaat ttccgaggag 2221 gacttatagt gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa 2281 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa 2341 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat 2401 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt 2461 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc 2521 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc 2581 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg 2641 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga 2701 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc 2761 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc 2821 tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat 2881 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg 2941 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac 3001 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa 3061 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag 3121 atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg 3181 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg 3241 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc 3301 taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa 3361 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc 3421 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac 3481 tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt 3541 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg 3601 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 3661 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 3721 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc 3781 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 3841 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt 3901 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc 3961 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg 4021 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg 4081 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa 4141 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct 4201 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga 4261 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc 4321 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac 4381 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc 4441 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact 4501 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga 4561 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg 4621 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga 4681 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga 4741 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg 4801 ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg 4861 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 4921 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt 4981 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 5041 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 5101 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 5161 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 5221 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 5281 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 5341 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 5401 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 5461 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 5521 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 5581 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 5641 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 5701 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt 5761 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 5821 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 5881 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 5941 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 6001 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 6061 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 6121 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 6181 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 6241 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 6301 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 6361 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 6421 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 6481 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 6541 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 6601 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 6661 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 6721 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 6781 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 6841 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 6901 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 6961 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 7021 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 7081 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 7141 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 7201 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 7261 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg 7321 gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc 7381 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg 7441 cttgaccgac aattgcatga agaatctgct tagg //