LOCUS Exported 7612 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pUSE-RapR-Src(KD)-myc SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7612) AUTHORS Karginov AV, Ding F, Kota P, Dokholyan NV, Hahn KM TITLE Engineered allosteric activation of kinases in living cells. JOURNAL Nat Biotechnol. 2010 Jun 27. ():. PUBMED 20581846 REFERENCE 2 (bases 1 to 7612) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7612 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 2841..2870 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" primer_bind complement(3202..3219) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3208..3432 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3478..3906 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3565..3584) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3775..3796 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3915..3935) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3920..4250 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4101..4236 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4163..4182 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4317..5111 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4371..4390) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4981..5000 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5285..5406 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5322..5341) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5376..5395 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5455..5471) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5455..5471) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5468..5490) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5479..5495 /label=lac operator /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-beta-D-thiogalactopyranoside (IPTG)." promoter complement(5503..5533) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5548..5569 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5686..5703) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5857..6445) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5937..5956) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6616..7476) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 7239..7258 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7477..7581) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gtttaaacgg gccctctaga ctcgagcggc cgccactgtg ctggatcatg ggcagcaaca 961 agagcaagcc caaggacgcc agccagcggc gccgcagcct ggagccctcg gaaaacgtgc 1021 acggggcagg gggcgccttc ccggcctcac agacaccgag caagcccgcc tccgccgacg 1081 gccaccgcgg gcccagcgcc gccttcgtgc cgcccgcggc cgagcccaag ctcttcggag 1141 gcttcaactc ctcggacacc gtcacctccc cgcagagggc ggggcctctg gcaggtgggg 1201 tgaccacctt tgtggccctc tatgactatg agtcacggac agagactgac ctgtccttca 1261 agaaagggga gcggctgcag attgttaaca acacggaggg agactggtgg ctggcacact 1321 cgctgagcac gggacagacc ggttacatcc ccagcaacta tgtggcgccc tccgactcca 1381 tccaggctga ggagtggtac tttggcaaga tcactagacg ggaatcagag cggctgctgc 1441 tcaacgccga gaacccgaga gggaccttcc tcgtgaggga gagtgagacc acaaaaggtg 1501 cctactgcct ctctgtatcc gacttcgaca atgccaaggg cctaaatgtg aaacactaca 1561 agatccgcaa gctggacagc ggcggtttct acatcacctc ccgcacccag ttcaacagcc 1621 tgcagcagct cgtggcttac tactccaaac atgctgatgg cctgtgtcac cgcctcacta 1681 ccgtatgtcc cacatccaag cctcagaccc agggattggc caaggatgcg tgggagatcc 1741 cccgggagtc cctgcggctg gaggtcaagc tgggccaggg ttgcttcgga gaggtgtgga 1801 tggggacctg gaacggacca ggtacctgcg tggtgcacta caccgggatg cttgaagatg 1861 gaaagaaatt tgattcctcc cgggacagaa acaagccctt taagtttatg ctaggcaagc 1921 aggaggtgat ccgaggctgg gaagaagggg ttgcccagat gagtgtgggt cagagagcca 1981 aactgactat atctccagat tatgcctatg gtgccactgg gcacccaggc atcatcccac 2041 cacatgccac tctcgtcttc gatgtggagc ttctaaaact ggaaggtccc ggaaccacga 2101 gggttgccat caaaactctg aagccaggca ccatgtcccc agaggccttc ctgcaggagg 2161 cccaagtcat gaagaaactg aggcacgaga aactggtgca gctgtatgct gtggtgtcgg 2221 aagaacccat ttacattgtg acagagtaca tgaacaaggg gagtctgctg gactttctca 2281 agggggaaac gggcaaatat ttgcggctac cccagctggt ggacatgtct gctcagatcg 2341 cttcaggcat ggcctatgtg gagcggatga actatgtgca ccgggacctt cgagccgcca 2401 atatcctagt aggggagaac ctggtgtgca aagtggccga ctttgggttg gcccggctca 2461 tagaagacaa cgaatacaca gcccggcaag gtgccaaatt ccccatcaag tggaccgccc 2521 ctgaagctgc tctgtacggc aggttcacca tcaagtcgga tgtgtggtcc tttgggattc 2581 tgctgaccga gctcaccact aagggaagag tgccctatcc tgggatggtg aaccgtgagg 2641 ttctggacca ggtggagcgg ggctaccgga tgccttgtcc ccccgagtgc cccgagtccc 2701 tgcatgacct tatgtgccag tgctggcgga aggagcccga ggagcggccc accttcgagt 2761 acctgcaggc cttcctggaa gactacttta cgtccactga gccacagtac cagcccgggg 2821 agaacctagg cggaccaggg gagcagaagc tgatctccga ggaggacctg tgaggactgt 2881 gtgcgcagaa ctgtcacctc tagagccaca gtaccagccc ggggagaacc tatagggact 2941 gtgtgcgcag aactgtcacc tctaggcttc cgggtgttgc ctccgatgca cctgcctccc 3001 ttcatcctct ctctggggct gcgttgcctc ggcaatgccc cttttggagc atggaagggc 3061 ttcagggatg gggcagcctg gaaggacagg tggaggctgg tacagtgact gtcccatgcc 3121 tgccatgatc tgcagaattc caccacactg gactagtgga tccgagctcg gtaccaagct 3181 taagtttaaa ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt 3241 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat 3301 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg 3361 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg 3421 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg 3481 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta 3541 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt 3601 tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg 3661 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat 3721 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac 3781 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag 3841 ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg 3901 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagg 3961 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 4021 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 4081 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 4141 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc 4201 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg 4261 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga 4321 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct 4381 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc 4441 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg 4501 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg 4561 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc 4621 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc 4681 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg 4741 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc 4801 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg 4861 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc 4921 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag 4981 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg 5041 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg 5101 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc 5161 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt 5221 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca 5281 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 5341 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 5401 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata 5461 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag 5521 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg 5581 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca 5641 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 5701 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 5761 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 5821 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 5881 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 5941 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 6001 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg 6061 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 6121 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 6181 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 6241 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac 6301 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 6361 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat 6421 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 6481 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 6541 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 6601 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 6661 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg 6721 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga 6781 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt 6841 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 6901 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt 6961 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 7021 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc 7081 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 7141 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 7201 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag 7261 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt 7321 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 7381 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 7441 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg 7501 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa 7561 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc //