LOCUS V020434 6947 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020434 VERSION V020434 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..2437 /label="CLK2(NM_001294338)" /note="CLK2(NM_001294338)" /gene="CLK2" CDS 2444..2509 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2547..2771 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2817..3245 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3259..3588 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3655..4449 /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 4623..4744 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4793..4809) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4817..4833 /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(4841..4871) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4886..4907 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5195..5780) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5951..6811) /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(6812..6916) /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 atgccgcatc ctcgaaggta 961 ccactcctca gagcgaggca gccgggggag ttaccgtgaa cactatcgga gccgaaagca 1021 taagcgacga agaagtcgct cctggtcaag tagtagtgac cggacacgac ggcgtcggcg 1081 agaggacagc taccatgtcc gttctcgaag cagttatgat gatcgttcgt ccgaccggag 1141 ggtgtatgac cggcgatact gtggcagcta cagacgcaac gattatagcc gggatcgggg 1201 agatgcctac tatgacacag actatcggca ttcctatgaa tatcagcggg agaacagcag 1261 ttaccgcagc cagcgcagca gccggaggaa gcacagacgg cggaggaggc gcagccggac 1321 atttagccgc tcatcttcgc agcacagcag ccggagagcc aagagtgtag aggacgacgc 1381 tgagggccac ctcatctacc acgtcgggga ctggctacaa gagcgatatg aaatcgttag 1441 caccttagga gaggggacct tcggccgagt tgtacaatgt gttgaccatc gcaggggtgg 1501 ggctcgagtt gccctgaaga tcattaagaa tgtggagaag tacaaggaag cagctcgact 1561 tgagatcaac gtgctagaga aaatcaatga gaaagaccct gacaacaaga acctctgtgt 1621 ccagatgttt gactggtttg actaccatgg ccacatgtgt atctcctttg agcttctggg 1681 ccttagcacc ttcgatttcc tcaaagacaa caactacctg ccctacccca tccaccaagt 1741 gcgccacatg gccttccagc tgtgccaggc tgtcaagttc ctccatgata acaagctgac 1801 acatacagac ctcaagcctg aaaatattct gtttgtgaat tcagactatg agctcaccta 1861 caacctagag aagaagcgag atgagcgcag tgtgaagagc acagctgtgc gggtggtaga 1921 ctttggcagt gccacctttg accatgagca ccatagcacc attgtctcca ctcgccatta 1981 ccgagcacca gaagtcatcc ttgagttggg ctggtcacag ccttgtgatg tgtggagtat 2041 aggctgcatc atctttgaat actatgtggg attcaccctc ttccagaccc atgacaacag 2101 agagcatcta gccatgatgg aaaggatctt gggtcctatc ccttcccgga tgatccgaaa 2161 gacaagaaag cagaaatatt tttaccgggg tcgcctggat tgggatgaga acacatcagc 2221 tgggcgctat gttcgtgaga actgcaaacc gctgcggcgg tatctgacct cagaggcaga 2281 ggaacaccac cagctcttcg atctgattga aagcatgcta gagtatgaac cagctaagcg 2341 gctgaccttg ggtgaagccc ttcagcatcc tttcttcgcc cgccttcggg ctgagccgcc 2401 caacaagttg tgggactcca gtcgggatat cagtcggctc gaggactaca aagaccatga 2461 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc 2521 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg 2581 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 2641 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 2701 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt 2761 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc 2821 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 2881 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 2941 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 3001 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc 3061 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 3121 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg 3181 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 3241 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca 3301 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca 3361 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc 3421 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 3481 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta 3541 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga 3601 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt 3661 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat 3721 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag 3781 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac 3841 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac 3901 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc 3961 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg 4021 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag 4081 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat 4141 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag 4201 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc 4261 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg 4321 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg 4381 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag 4441 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat 4501 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 4561 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc 4621 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 4681 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 4741 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 4801 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 4861 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 4921 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 4981 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 5041 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 5101 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 5161 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 5221 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 5281 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 5341 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 5401 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 5461 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 5521 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 5581 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 5641 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 5701 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc 5761 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 5821 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 5881 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 5941 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 6001 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 6061 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 6121 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 6181 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 6241 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 6301 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 6361 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 6421 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 6481 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 6541 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 6601 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 6661 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 6721 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 6781 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 6841 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 6901 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc //