LOCUS V026532 6851 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V026532 VERSION V026532 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 941..2341 /label="Rora(NM_001289916)" /note="Rora(NM_001289916)" /gene="Rora" CDS 2348..2413 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2451..2675 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2721..3149 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3163..3492 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3559..4353 /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 4527..4648 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4697..4713) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4721..4737 /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(4745..4775) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4790..4811 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5099..5684) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5855..6715) /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(6716..6820) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgtattttg tgatcgcagc 961 gatgaaagct caaattgaaa ttattccatg caagatctgt ggagacaaat cgtcaggaat 1021 ccattatggt gtcattacgt gtgaaggctg caagggcttt ttcaggagaa gtcagcagag 1081 caatgccacc tactcctgtc ctcgtcagaa gaactgtttg attgatcgga ccagcagaaa 1141 ccgctgccag cattgtcggc tgcagaaatg cctggccgtg gggatgtctc gagatgctgt 1201 caagtttggt cggatgtcca agaagcagag agacagcttg tacgccgagg tgcagaagca 1261 ccggatgcag cagcagcagc gagaccacca gcagcagcct ggggaggcgg agccgctgac 1321 gcccacctac aacatctcag ccaatgggct gacggaactg catgatgacc tcagcaccta 1381 tatggatggg cacacccccg agggcagcaa ggccgactca gccgtcagca gcttctacct 1441 ggacatccag ccctccccag accagtcggg attggacatc aatgggatca aacccgaacc 1501 catatgtgac tacacaccag catctggctt cttcccctac tgttccttca ccaacggaga 1561 gacttcccca accgtgtcca tggcagaact agaacacctt gcccagaaca tatccaaatc 1621 ccacctggaa acctgccagt acttgcggga agagctccag cagataacgt ggcagacctt 1681 cctgcaggag gagattgaaa actaccagaa caagcagaga gaggtgatgt ggcagctgtg 1741 tgccatcaag attacagaag ctatccagta tgtggtggag tttgccaaac gcattgatgg 1801 atttatggag ctgtgtcaaa atgatcaaat tgtgcttcta aaagcaggct cgctagaggt 1861 ggtgtttatt aggatgtgcc gtgcctttga ctctcagaac aacaccgtgt actttgacgg 1921 gaagtatgcg agccccgatg tcttcaagtc cctaggttgt gaagacttca tcagctttgt 1981 gtttgaattt gggaagagtt tgtgttctat gcacctgacc gaagacgaaa tcgcgttatt 2041 ttctgcattc gtactgatgt cagcggatcg ctcgtggctt caggaaaagg taaaaataga 2101 aaagctgcaa cagaaaattc agctggccct tcagcacgtc ctacagaaga accaccgaga 2161 agatggaatt ctaaccaagc taatatgcaa ggtgtctacg ttaagagccc tatgtggacg 2221 acatacggaa aagctaatgg catttaaagc aatataccca gacattgtgc gactccattt 2281 tcctccatta tacaaggaat tgttcacttc agaatttgag ccagccatgc aaatcgatgg 2341 gctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga 2401 tgacgatgac aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc 2461 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc 2521 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg 2581 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa 2641 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg 2701 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt 2761 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt 2821 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct 2881 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg 2941 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga 3001 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc 3061 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga 3121 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt 3181 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 3241 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 3301 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 3361 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 3421 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 3481 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca 3541 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct 3601 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc 3661 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc 3721 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc 3781 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg 3841 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc 3901 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac 3961 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat 4021 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc 4081 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg 4141 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg 4201 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc 4261 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc 4321 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg 4381 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa 4441 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc 4501 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat 4561 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 4621 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga 4681 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 4741 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 4801 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 4861 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 4921 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 4981 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 5041 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 5101 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 5161 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 5221 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 5281 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 5341 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 5401 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 5461 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 5521 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct 5581 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt 5641 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 5701 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 5761 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 5821 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 5881 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 5941 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 6001 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 6061 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 6121 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 6181 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 6241 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 6301 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 6361 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 6421 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 6481 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 6541 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 6601 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 6661 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 6721 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 6781 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 6841 cacctgacgt c //