LOCUS V035329 6848 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035329 VERSION V035329 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 955..2748 /label="NR4A2(NM_006186)" /note="NR4A2(NM_006186)" /gene="NR4A2" CDS 2770..2799 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" CDS 2815..2832 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" polyA_signal 2861..3085 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3131..3559 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3573..3903 /note="SV40 enhancer and early promoter" /label="SV40 promoter" promoter 3951..3998 /note="synthetic bacterial promoter " /label="EM7 promoter" CDS 4017..4391 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /note="confers resistance to bleomycin, phleomycin, and Zeocin™" /product="antibiotic-binding protein" /transl_table=1 /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD*" /label="BleoR" polyA_signal 4521..4642 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4691..4707) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4715..4731 /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(4739..4769) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4784..4805 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5093..5681) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5852..6712) /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(6713..6817) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt 901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattcgc caccatgcct 961 tgtgttcagg cgcagtatgg gtcctcgcct caaggagcca gccccgcttc tcagagctac 1021 agttaccact cttcgggaga atacagctcc gatttcttaa ctccagagtt tgtcaagttt 1081 agcatggacc tcaccaacac tgaaatcact gccaccactt ctctccccag cttcagtacc 1141 tttatggaca actacagcac aggctacgac gtcaagccac cttgcttgta ccaaatgccc 1201 ctgtccggac agcagtcctc cattaaggta gaagacattc agatgcacaa ctaccagcaa 1261 cacagccacc tgccccccca gtctgaggag atgatgccgc actccgggtc ggtttactac 1321 aagccctcct cgcccccgac gcccaccacc ccgggcttcc aggtgcagca cagccccatg 1381 tgggacgacc cgggatctct ccacaacttc caccagaact acgtggccac tacgcacatg 1441 atcgagcaga ggaaaacgcc agtctcccgc ctctccctct tctcctttaa gcaatcgccc 1501 cctggcaccc cggtgtctag ttgccagatg cgcttcgacg ggcccctgca cgtccccatg 1561 aacccggagc ccgccggcag ccaccacgtg gtggacgggc agaccttcgc tgtgcccaac 1621 cccattcgca agcccgcgtc catgggcttc ccgggcctgc agatcggcca cgcgtctcag 1681 ctgctcgaca cgcaggtgcc ctcaccgccg tcgcggggct ccccctccaa cgaggggctg 1741 tgcgctgtgt gtggggacaa cgcggcctgc caacactacg gcgtgcgcac ctgtgagggc 1801 tgcaaaggct tctttaagcg cacagtgcaa aaaaatgcaa aatacgtgtg tttagcaaat 1861 aaaaactgcc cagtggacaa gcgtcgccgg aatcgctgtc agtactgccg atttcagaag 1921 tgcctggctg ttgggatggt caaagaagtg gttcgcacag acagtttaaa aggccggaga 1981 ggtcgtttgc cctcgaaacc gaagagccca caggagccct ctcccccttc gcccccggtg 2041 agtctgatca gtgccctcgt cagggcccat gtcgactcca acccggctat gaccagcctg 2101 gactattcca ggttccaggc gaaccctgac tatcaaatga gtggagatga cacccagcat 2161 atccagcaat tctatgatct cctgactggc tccatggaga tcatccgggg ctgggcagag 2221 aagatccctg gcttcgcaga cctgcccaaa gccgaccaag acctgctttt tgaatcagct 2281 ttcttagaac tgtttgtcct tcgattagca tacaggtcca acccagtgga gggtaaactc 2341 atcttttgca atggggtggt cttgcacagg ttgcaatgcg ttcgtggctt tggggaatgg 2401 attgattcca ttgttgaatt ctcctccaac ttgcagaata tgaacatcga catttctgcc 2461 ttctcctgca ttgctgccct ggctatggtc acagagagac acgggctcaa ggaacccaag 2521 agagtggaag aactgcaaaa caagattgta aattgtctca aagaccacgt gactttcaac 2581 aatggggggt tgaaccgccc caattatttg tccaaactgt tggggaagct cccagaactt 2641 cgtacccttt gcacacaggg gctacagcgc attttctacc tgaaattgga agacttggtg 2701 ccaccgccag caataattga caaacttttc ctggacactt tacctttcct cgagtctaga 2761 gggcccttcg aacaaaaact catctcagaa gaggatctga atatgcatac cggtcatcat 2821 caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc tagttgccag 2881 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact 2941 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt 3001 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat 3061 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg 3121 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 3181 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 3241 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg 3301 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca 3361 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc 3421 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct 3481 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa 3541 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc 3601 caggctcccc aggcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg 3661 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 3721 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 3781 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 3841 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc 3901 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt 3961 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg 4021 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt 4081 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg 4141 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca 4201 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg 4261 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt 4321 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg 4381 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg 4441 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc 4501 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca 4561 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 4621 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg 4681 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 4741 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 4801 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 4861 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 4921 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 4981 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 5041 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 5101 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 5161 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 5221 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 5281 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 5341 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 5401 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 5461 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 5521 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 5581 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 5641 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 5701 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 5761 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 5821 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 5881 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 5941 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 6001 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 6061 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 6121 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 6181 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 6241 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 6301 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 6361 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 6421 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 6481 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 6541 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 6601 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 6661 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 6721 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 6781 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 6841 ctgacgtc //