LOCUS V024758 7084 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024758 VERSION V024758 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..2577 /label="Kcnn1(NM_001363407)" /note="Kcnn1(NM_001363407)" /gene="Kcnn1" CDS 2587..2616 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" CDS 2623..2646 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" polyA_signal 2684..2908 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2954..3382 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3396..3725 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3792..4586 /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 4760..4881 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4930..4946) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4954..4970 /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(4978..5008) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5023..5044 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5332..5917) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6088..6948) /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(6949..7053) /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 accatgagta gccacagcca caatggcagc gtggggcagc ctctgggcag cgggcctgga 1021 ttcctgggct gggagcctgt ggaccctgag gcaggccgcc ccctgcagcc cacccaaggc 1081 ccaggcctgc agatggtggc caagggtcag cctgtcaggc tgtcacccgg tggttccagg 1141 ggccaccccc aggagcagga ggaggaagag gaagaggagg aggaggagga ggacaagaca 1201 ggctcaggga agcccccaac agtcagccac cgcctgggac accgcagggc cctctttgaa 1261 aagcgtaaac ggctcagtga ctatgcgctc atctttggca tgtttgggat tgtcgtcatg 1321 gtgacagaaa cagagctgtc ctggggtgta tacaccaagg agtcactctg ctcttttgct 1381 ctgaaatgcc tcatcagcct gtccactgtc atcttgcttg gccttgtcat cctgtaccac 1441 gcccgagaga tccagctgtt cttggtggac aatggtgccg acgactggcg tatcgccatg 1501 acgtgggagc gcgtgtccct gatctcgctg gagttggtcg tgtgtgccat ccacccggtg 1561 cccggccact atcgcttcac gtggacggca cgactggcct tctctctggt gccgtcggca 1621 gccgaggcag acctggatgt gctgctgtcc atccccatgt tcctgcgcct ctacctgctg 1681 gctcgggtca tgctcctgca cagccgcatc ttcaccgatg catccagccg cagcatcggg 1741 gcccttaacc gcgtcacctt caacacacgc ttcgtcacca agacgctcat gaccatctgc 1801 cccggcactg tgttgttggt cttcagcgtc tcctcctgga tcgttgctgc gtggacagtg 1861 cgcgtgtgtg agaggtacca cgataagcag gaagtgacca gcaacttcct gggagccatg 1921 tggctcatct ccatcacctt cttgtccatt ggctatggag acatggtgcc gcatacctac 1981 tgtgggaagg gtgtgtgtct gctcactggc atcatgggag caggctgcac tgcactcgtg 2041 gtggccgtcg tggctcggaa gttggaactc accaaggctg agaaacacgt gcacaacttc 2101 atgatggaca cacagctcac caagcgggtc aaaaatgctg ctgcaaacgt tctcagggag 2161 acatggctca tctacaaaca caccaggctg gtgaagaagc cagaccaagg ccgggttcgg 2221 aaacaccagc gtaagttcct tcaggccatc catcaggctc agaagctccg aagtgtgaag 2281 attgaacaag ggaaggtgaa cgatcaggcc aacacgctgg ctgagctggc caaggcacag 2341 agcatcgcat atgaggtggt gtcagagctg caggcccagc aggaggagtt ggaggcacgc 2401 ctagccgcct tggagagccg actggatgtc ctgggtgcct ccctgcaggc tctaccaggc 2461 cttatagccc aagccatatg ccctctacca ccaccctggc ctgggcctgg tcacctggcc 2521 acagccaccc agagcccaca aagccactgg ctgcccacca tgggatcaga ctgtgggtga 2581 gatatcgagc agaaactcat ctcagaagag gatctgctcg aggattacaa ggatgacgac 2641 gataagtagt gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc 2701 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc 2761 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct 2821 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg 2881 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct 2941 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 3001 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 3061 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 3121 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 3181 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 3241 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 3301 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 3361 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt 3421 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 3481 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 3541 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 3601 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 3661 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 3721 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag 3781 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 3841 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 3901 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 3961 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 4021 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 4081 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 4141 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 4201 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 4261 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc 4321 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 4381 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 4441 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 4501 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 4561 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc 4621 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg 4681 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct 4741 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa 4801 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 4861 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc 4921 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 4981 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 5041 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 5101 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 5161 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 5221 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 5281 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 5341 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 5401 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 5461 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 5521 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 5581 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 5641 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 5701 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 5761 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 5821 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg 5881 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 5941 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 6001 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 6061 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 6121 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 6181 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 6241 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 6301 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 6361 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 6421 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 6481 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 6541 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 6601 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 6661 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 6721 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 6781 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 6841 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 6901 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 6961 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 7021 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga 7081 cgtc //