LOCUS V026134 6722 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V026134 VERSION V026134 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 202..581 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 582..785 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 830..848 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 956..979 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 980..2242 /label="Htr1a(NM_008308)" /note="Htr1a(NM_008308)" /gene="Htr1a" polyA_signal 2289..2513 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2559..2987 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3001..3330 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3397..4191 /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 4365..4486 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4535..4551) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4559..4575 /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(4583..4613) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4628..4649 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4937..5522) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5693..6553) /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(6554..6658) /gene="bla" /label="AmpR promoter" ORIGIN 1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg 181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta 241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc 841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc 901 cgccaccatg aagaccatca tcgctctgag ctacatcttc tgcctggtct tcgctgacta 961 caaggacgac gacgacaaga tggatatgtt cagtcttggc cagggcaaca acaccacaac 1021 gtccctggag cccttcggga caggcggcaa cgatactggc ctctccaacg tgaccttcag 1081 ctaccaagtg atcacctctc ttttgctggg gacgctcatt ttctgcgcgg tgctcggcaa 1141 tgcctgcgtg gttgctgcca tcgccctgga gcgctccctt cagaatgttg ccaactatct 1201 catcggctcc ttggcggtca ccgatctcat ggtgtcagtg ctggtgctgc ccatggctgc 1261 tctgtaccag gtgctcaaca agtggactct gggtcaggtc acctgtgacc tgtttatcgc 1321 cctggatgtg ctgtgctgca cttcgtccat cctgcacctg tgcgccatcg cgctagacag 1381 gtactgggca atcaccgacc ctatagacta cgtgaacaag aggacgcccc ggcgcgccgc 1441 tgcgctgatc tcgctcactt ggctcattgg ctttctcatc tccatcccgc ctatgctggg 1501 ctggcgcacc ccggaagacc gctcgaaccc caacgagtgc accatcagca aggaccacgg 1561 gtacaccatc tactccactt tcggcgcttt ctatattccg ctgctgctca tgctggtcct 1621 ctatgggcgc atcttcagag ccgcgcgctt ccgaatccgc aagacggtca agaaggtgga 1681 aaagaaggga gcgggcacca gcttcggaac atcgtcggcc ccgcccccca agaagagcct 1741 gaatggtcag ccaggtagtg gggactgcag gcgcagtgct gagaacaggg cggtggggac 1801 tccgtgcgct aatggggcgg tgagacaggg tgaggacgac gccaccctgg aggtgatcga 1861 ggtgcatcga gtgggcaact ccaaagggca ccttcctctg cccagcgaat caggagctac 1921 ctcctatgtc cccgcctgct tggagagaaa aaatgagcgc actgctgagg caaagcgcaa 1981 gatggccttg gcccgtgaga ggaagacagt gaagacactg ggcatcatca tgggcacctt 2041 catcctctgc tggctgccct ttttcattgt ggccctggtc ctacctttct gtgagagcag 2101 ttgccacatg cctgagttgt tgggtgccat aattaactgg ctaggctact ccaactccct 2161 gctcaaccca gttatttatg cttatttcaa caaagacttt caaaacgctt ttaagaagat 2221 catcaagtgc aagttctgcc gctgactcga gtctagaggg cccgtttaaa cccgctgatc 2281 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc 2341 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc 2401 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg 2461 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga 2521 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt 2581 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc 2641 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca 2701 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc 2761 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt 2821 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac 2881 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc 2941 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat 3001 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 3061 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga 3121 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc 3181 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt 3241 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga 3301 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc 3361 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac 3421 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca 3481 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt 3541 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg 3601 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga 3661 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct 3721 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg 3781 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg 3841 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc 3901 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat 3961 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac 4021 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt 4081 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct 4141 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc 4201 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca 4261 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga 4321 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag 4381 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 4441 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac 4501 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt 4561 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg 4621 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt 4681 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt 4741 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc 4801 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg 4861 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg 4921 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac 4981 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 5041 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 5101 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg 5161 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct 5221 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac 5281 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt 5341 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc 5401 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca 5461 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 5521 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 5581 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 5641 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 5701 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 5761 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 5821 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 5881 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 5941 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 6001 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 6061 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 6121 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 6181 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 6241 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 6301 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 6361 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 6421 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 6481 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 6541 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 6601 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 6661 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg 6721 gt //