LOCUS V020365 7805 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020365 VERSION V020365 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..2485 /label="Nr4a1(NM_010444)" /note="Nr4a1(NM_010444)" /gene="Nr4a1" CDS 2492..2557 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2581..3169 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3235..3283 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3485..3913 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3927..4256 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4323..5117 /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 5293..5414 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5463..5479) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5487..5503 /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(5511..5541) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5556..5577 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5865..6453) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6624..7484) /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(7485..7589) /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 ccatgccctg tattcaagct caatatggaa caccagcaac 721 gagcccagga ccgcgtgacc acctgaccgg tgatcccctg gcccttgagt tcggcaagcc 781 taccatggac ctggccagcc ccgagacagc acctgccgca cctgctacac tgcccagctt 841 cagcaccttc atggacgggt acaccggaga gtttgacacc ttcctctacc agctgccggg 901 gacgacccag ccgtgctcct cagcttgttc ctctgcctcc tccacgtctt cttcctcatc 961 ctcggccacc tcccccgctt cggcgtcctt caagtttgag gacttccagg tgtacggctg 1021 ctacccgggc accctgagcg gcccattaga tgagacccta tcctccagcg gctctgagta 1081 ctatggcagt ccctgctcag ccccctcgcc atctacaccc aacttccagc cgtcccagct 1141 ttctccctgg gacggctcat ttggccactt ctccccgagc cagacttatg aaggcctctg 1201 ggcatggaca gagcagttgc ctaaggcttc ttcagggcct ccgccacctc caaccttctt 1261 ctccttcagt cctcccactg gccccagccc cagcctggcc cagagttctc tgaaattgtt 1321 cccaccacca gccacccacc agcttgggga gggggagagc tattccatgc cagcagcttt 1381 ccccggcttg gcacccacct ctccgaaccg tgacacttcc ggcattctgg acgcacccgt 1441 gacctccacc aagtcccgga gcggggcttc aggtggcagc gagggccgct gtgcagtctg 1501 tggtgacaat gcttcgtgtc agcactatgg ggtccgcacc tgtgagggct gcaagggctt 1561 cttcaagcgc acagtacaga aaagcgccaa gtacatctgc ctggcaaaca aggattgccc 1621 tgtggacaag aggcggcgga accgctgcca gttctgccgc ttccagaagt gcctggctgt 1681 gggcatggtg aaggaagttg tacggacaga cagcctaaaa gggcggcggg gccggctacc 1741 ttcaaaaccc aagcagcctc cagatgcctc ccctaccaat cttctcactt ccctcatccg 1801 ggcacacttg gactccgggc ctagcactgc caaattggac tattccaagt tccaggaact 1861 ggtgctgccc cgcttcggga aggaagatgc cggtgacgtg caacaatttt atgacttgct 1921 ctctggttcc ctggacgtta tccgaaagtg ggcagaaaaa atccctggct tcattgagct 1981 ttgcccagga gaccaagacc tgttgctaga gtctgccttc ctggaactct tcatcctccg 2041 cctggcatac cgatctaaac ccggtgaggg gaagctcatc ttctgctcag gcctggtact 2101 acaccagctg cagtgtgccc gtggctttgg tgattggatt gacaacatcc tggccttctc 2161 acggtccctg cacagcttgg gtgttgatgt tcccgccttt gcctgcctgt ccgctctggt 2221 cctcatcact gatcgacacg ggctccagga ccctcgtcgg gtggaagagc tgcagaatcg 2281 cattgctagc tgtctgaagg agcacatggc taccgtggca ggagacccac agccggccag 2341 ctgcctgtca cgtctgctgg gcaaactgcc tgagcttcgg accctgtgca ctcaaggcct 2401 gcagcgcatc ttttgcctca agttggagga cttggtaccc cctccaccta ttgtggacaa 2461 gatctttatg gacacattgt ctttcctcga ggactacaaa gaccatgacg gtgattataa 2521 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg atatctcgac 2581 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 2641 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 2701 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 2761 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 2821 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 2881 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 2941 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc 3001 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 3061 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt 3121 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg 3181 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa 3241 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg 3301 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc 3361 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc 3421 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat 3481 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 3541 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 3601 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga 3661 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 3721 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 3781 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 3841 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 3901 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 3961 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 4021 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 4081 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 4141 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct 4201 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 4261 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc 4321 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 4381 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 4441 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 4501 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 4561 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 4621 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 4681 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 4741 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 4801 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg 4861 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 4921 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 4981 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 5041 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 5101 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc 5161 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga 5221 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc 5281 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 5341 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 5401 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca 5461 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 5521 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 5581 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 5641 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 5701 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 5761 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 5821 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 5881 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 5941 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 6001 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa 6061 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 6121 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 6181 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 6241 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 6301 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 6361 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 6421 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 6481 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 6541 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 6601 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 6661 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 6721 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 6781 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 6841 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 6901 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 6961 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 7021 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 7081 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 7141 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 7201 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 7261 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 7321 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 7381 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 7441 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 7501 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 7561 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc 7621 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 7681 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 7741 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 7801 ttagg //