LOCUS V024479 6524 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024479 VERSION V024479 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..2014 /label="Mapk1(NM_011949)" /note="Mapk1(NM_011949)" /gene="Mapk1" CDS 2021..2086 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2124..2348 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2394..2822 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2836..3165 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3232..4026 /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 4200..4321 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4370..4386) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4394..4410 /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(4418..4448) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4463..4484 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4772..5357) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5528..6388) /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(6389..6493) /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 atggcggcgg cggcggcggc 961 gggcccggag atggtccgcg ggcaggtgtt cgacgtaggg ccgcgctaca ccaacctctc 1021 gtacatcgga gaaggcgcct acggcatggt ttgctctgct tatgataatc tcaacaaagt 1081 tcgagttgct atcaagaaaa tcagtccttt tgagcaccag acctactgtc aaagaaccct 1141 aagagagata aaaatcttac tgcgcttcag acatgagaac atcattggca tcaatgacat 1201 catccgggca ccaaccattg agcaaatgaa agatgtatat atagtacagg acctcatgga 1261 gacggacctt tacaagctct tgaagacaca gcacctcagc aatgaccaca tctgctattt 1321 tctttatcag atcctgagag ggctaaagta tatccattca gctaacgttc tgcaccgtga 1381 cctcaagcct tccaacctcc tgctgaacac cacttgtgat ctcaagatct gtgactttgg 1441 ccttgcccgt gttgcagatc cagatcatga tcacacaggg ttcttgacag agtacgtagc 1501 cacacgttgg tacagagctc cagaaattat gttgaattcc aagggttata ccaagtccat 1561 tgatatttgg tctgtgggct gcatcctggc agagatgcta tccaacaggc ctatcttccc 1621 aggaaagcat taccttgacc agctgaatca catcctgggt attcttggat ctccatcaca 1681 ggaagatctg aattgtataa taaatttaaa agctagaaac tatttgcttt ctctcccgca 1741 caaaaataag gtgccatgga acaggttgtt cccaaatgct gactccaaag ctctggattt 1801 actggataaa atgttgacat ttaaccctca caagaggatt gaagttgaac aggctctggc 1861 ccacccatac ctggagcagt attatgaccc aagtgatgag cccattgctg aagcgccatt 1921 caagtttgac atggagttgg acgacttacc taaggagaag ctcaaagaac tcatttttga 1981 agagactgct agattccagc caggatacag atctctcgag gactacaaag accatgacgg 2041 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt 2101 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc 2161 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa 2221 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg 2281 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg 2341 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc 2401 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 2461 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 2521 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 2581 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 2641 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 2701 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 2761 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 2821 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc 2881 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc 2941 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg 3001 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 3061 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc 3121 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct 3181 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa 3241 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac 3301 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg 3361 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag 3421 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt 3481 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg 3541 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg 3601 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga 3661 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag 3721 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat 3781 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt 3841 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg 3901 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt 3961 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc 4021 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac 4081 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 4141 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 4201 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 4261 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 4321 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 4381 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 4441 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 4501 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 4561 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 4621 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 4681 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 4741 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 4801 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 4861 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 4921 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 4981 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 5041 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 5101 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 5161 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 5221 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 5281 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca 5341 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 5401 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 5461 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 5521 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 5581 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 5641 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 5701 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 5761 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 5821 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 5881 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 5941 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 6001 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 6061 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 6121 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 6181 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 6241 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 6301 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 6361 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 6421 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 6481 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc //