LOCUS V024929 8017 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024929 VERSION V024929 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 964..3510 /label="Kcnn2(NM_001312905)" /note="Kcnn2(NM_001312905)" /gene="Kcnn2" CDS 3520..3549 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" CDS 3556..3579 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" polyA_signal 3617..3841 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3887..4315 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4329..4658 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4725..5519 /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 5693..5814 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5863..5879) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5887..5903 /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(5911..5941) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5956..5977 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6265..6850) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7021..7881) /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(7882..7986) /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 atccactagt ccagtgtggt ggaattcgcc 961 accatgttcg ggagaacgtg caaatgtgat gtcatgccca ttgttttggt gcgcccaacc 1021 aatcggactc gccgcctgga ttctaccgga gccggcatgg gcccttcctc gcaccagcag 1081 caggagtccc cgctcccgac cataacgcat tgcgcagggt gcaccaccgc ctggtctccc 1141 tgcagcttta acagctctga catggaaacc ccattgcagt tccagcgcgg cttcttccca 1201 gagcagccgc cgccaccgcc gcgctcctca cacctgcatt gccagcagca gcaacagagc 1261 caggacaagc cgtgcgcgcc cttcgcgccc ctcccgcacc ctcaccacca cccgcacctc 1321 gcgcaccagc agccgggcag cggtggcagc agcccatgcc tccggtgcaa cagctgcgcc 1381 tcctccggtg ccccggcagc gggggcgggg gcgggggata acctgtccct gctgctccgc 1441 acctcatcgc ccggcggcgc cttccggacc cgcacctcct cgccactgtc gggctcttcg 1501 tgctgttgct gctgctcgtc gcgccggggc agccagctca atgtgagcga gctgacgccg 1561 tccagccatg ccagtgcgct ccggcagcag tacgcgcagc agcccgcgtc cgcctcccag 1621 taccaccagt gccacagcct gcagcccgcc accagcccca caggcagcct gggcagcctg 1681 ggctccgggc ccccgctctc gcaccaccac caccacccgc acccggcgca ccaccagcat 1741 caccaacccc aggcgcgccg cgagagcaac cccttcaccg aaatagccat gagcagctgc 1801 aggtacaacg ggggcgtcat gcgtccgctc agcaacttga gctcgtcccg ccggaacctg 1861 caggagatgg actccgaggc tcagcccctg cagccccccg cgtccgtcgt aggaggaggt 1921 ggtggtgcgt cctccccgtc tgctgccgcc gccgcctcat cctcagcccc ggagatcgtg 1981 gtgtctaagc cggagcacaa caattctaac aacctggcgc tctacgggac tggcggcgga 2041 ggcagcaccg gaggcggcgg cggcggcagc gggcatggca gcagcagcgg caccaagtcc 2101 agcaaaaaga agaaccagaa catcggctac aagctgggcc atcggcgcgc cctgtttgag 2161 aagcgcaagc ggctcagcga ctatgcgctt atcttcggca tgttcggcat cgtggttatg 2221 gtcattgaga ccgagctgtc ttggggcgcc tacgacaagg cgtcgctgta ttctttagct 2281 ctgaaatgcc ttatcagtct ctccacgatc atcctgcttg gtctgatcat cgtgtaccac 2341 gccagggaaa tacagttatt catggtggac aatggagcag atgactggag aatagccatg 2401 acttatgagc gtattttctt catctgcttg gaaatactgg tgtgtgctat ccatcccata 2461 cctgggaatt atacgttcac atggacagcc cggcttgcct tctcctatgc cccctccaca 2521 accaccgcag atgtggatat tattttatct ataccaatgt tcttaaggct ctatctgatt 2581 gccagagtca tgctattaca tagcaaactt ttcaccgatg cctcgtctag aagcattggg 2641 gcacttaata agataaactt caatacgcgt tttgttatga agactttaat gactatctgc 2701 ccaggaactg tgctcttggt ttttagtatc tcattatgga taattgccgc atggactgtc 2761 cgagcttgtg aaaggtacca tgatcaacag gatgtcacta gcaacttcct tggagcaatg 2821 tggttgatat caataacttt tctctccatt ggttatggtg acatggtacc caacacatac 2881 tgtgggaaag gagtctgctt gcttactgga atcatgggtg caggttgcac agccctggtg 2941 gtagccgtag tggcaagaaa gctagaactt accaaagcag aaaagcatgt gcacaatttc 3001 atgatggata ctcagctgac caaaagagtg aaaaacgcag ccgccaatgt actcagggaa 3061 acgtggttaa tctacaaaaa cacaaagcta gtgaaaaaga tcgaccacgc aaaagtaaga 3121 aagcatcaac ggaaattctt gcaagctatt catcaattaa gaagtgtgaa gatggaacag 3181 aggaaactga atgaccaagc aaatacccta gtggatctgg caaagaccca gaatatcatg 3241 tatgacatga tttccgactt gaatgaaagg agtgaagact ttgagaaaag gatcgtcacc 3301 ctggaaacaa aactagagac tttgattggt agcattcatg ccctccctgg gctaatcagc 3361 cagaccatca gacagcagca aagggacttc attgagacac agatggagaa ctacgacaag 3421 catgtctcct acaatgctga gcgctcccgg tcctcgtcca ggaggcggcg gtcctcctcc 3481 acagcgccac caacttcatc agagagtagc taggatatcg agcagaaact catctcagaa 3541 gaggatctgc tcgaggatta caaggatgac gacgataagt agtgagggcc cgtttaaacc 3601 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 3661 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 3721 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 3781 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 3841 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc 3901 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 3961 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 4021 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac 4081 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 4141 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 4201 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg 4261 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc 4321 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 4381 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 4441 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 4501 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 4561 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 4621 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat 4681 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg 4741 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 4801 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 4861 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 4921 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 4981 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 5041 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 5101 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 5161 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 5221 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 5281 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 5341 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 5401 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 5461 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 5521 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt 5581 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg 5641 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt 5701 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 5761 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 5821 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 5881 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 5941 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 6001 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 6061 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 6121 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 6181 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 6241 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 6301 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 6361 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 6421 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 6481 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 6541 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 6601 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 6661 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 6721 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 6781 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 6841 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 6901 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 6961 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 7021 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 7081 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 7141 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 7201 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 7261 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 7321 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 7381 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 7441 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 7501 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 7561 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 7621 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 7681 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 7741 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 7801 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 7861 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 7921 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 7981 tccgcgcaca tttccccgaa aagtgccacc tgacgtc //