LOCUS Exported 7140 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3-MTS-CA-c-Src-FLAG SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7140) AUTHORS Ogura M, Yamaki J, Homma MK, Homma Y TITLE Mitochondrial c-Src regulates cell survival through phosphorylation of respiratory chain components. JOURNAL Biochem J. 2012 Oct 15;447(2):281-9. PUBMED 22823520 REFERENCE 2 (bases 1 to 7140) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7140 /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 2659..2682 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter complement(2692..2710) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2693..2710) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2730..2747) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2736..2960 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3006..3434 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3093..3112) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3303..3324 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3443..3463) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3448..3778 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3629..3764 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3691..3710 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3845..4639 /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(3899..3918) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4509..4528 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4813..4934 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4850..4869) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4904..4923 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4983..4999) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4983..4999) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4996..5018) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5007..5023 /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(5031..5061) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5076..5097 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5214..5231) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5385..5973) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5465..5484) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6144..7004) /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 6767..6786 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7005..7109) /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 gcttgccacc 901 atgcaggcgg ctcggatggc cgcgagcttg gggcggcagc tgctgaggct cgggggcgga 961 agctcgcggc tcacggcgct cctggggcag ccccggcccg gccctgcccg gcggccctat 1021 gccgggggta ccgagctcgg atctatgggt agcaacaaga gcaagcccaa ggatgccagc 1081 cagcggcgcc gcagcctgga gcccgccgag aacgtgcacg gcgctggcgg gggcgctttc 1141 cccgcctcgc agacccccag caagccagcc tcggccgacg gccaccgcgg ccccagcgcg 1201 gccttcgccc ccgcggccgc cgagcccaag ctgttcggag gcttcaactc ctcggacacc 1261 gtcacctccc cgcagagggc gggcccgctg gccggtggag tgaccacctt tgtggccctc 1321 tatgactatg agtctaggac ggagacagac ctgtccttca agaaaggcga gcggctccag 1381 attgtcaaca acacagaggg agactggtgg ctggcccact cgctcagcac aggacagaca 1441 ggctacatcc ccagcaacta cgtggcgccc tccgactcca tccaggctga ggagtggtat 1501 tttggcaaga tcaccagacg ggagtcagag cggttactgc tcaatgcaga gaacccgaga 1561 gggaccttcc tcgtgcgaga aagtgagacc acgaaaggtg cctactgcct ctcagtgtct 1621 gacttcgaca acgccaaggg cctcaacgtg aagcactaca agatccgcaa gctggacagc 1681 ggcggcttct acatcacctc ccgcacccag ttcaacagcc tgcagcagct ggtggcctac 1741 tactccaaac acgccgatgg cctgtgccac cgcctcacca ccgtgtgccc cacgtccaag 1801 ccgcagactc agggcctggc caaggatgcc tgggagatcc ctcgggagtc gctgcggctg 1861 gaggtcaagc tgggccaggg ctgctttggc gaggtgtgga tggggacctg gaacggtacc 1921 accagggtgg ccatcaaaac cctgaagcct ggcacgatgt ctccagaggc cttcctgcag 1981 gaggcccagg tcatgaagaa gctgaggcat gagaagctgg tgcagttgta tgctgtggtt 2041 tcagaggagc ccatttacat cgtcacggag tacatgagca aggggagttt gctggacttt 2101 ctcaaggggg agacaggcaa gtacctgcgg ctgcctcagc tggtggacat ggctgctcag 2161 atcgcctcag gcatggcgta cgtggagcgg atgaactacg tccaccggga ccttcgtgca 2221 gccaacatcc tggtgggaga gaacctggtg tgcaaagtgg ccgactttgg gctggctcgg 2281 ctcattgaag acaatgagta cacggcgcgg caaggtgcca aattccccat caagtggacg 2341 gctccagaag ctgccctcta tggccgcttc accatcaagt cggacgtgtg gtccttcggg 2401 atcctgctga ctgagctcac cacaaaggga cgggtgccct accctgggat ggtgaaccgc 2461 gaggtgctgg accaggtgga gcggggctac cggatgccct gcccgccgga gtgtcccgag 2521 tccctgcacg acctcatgtg ccagtgctgg cggaaggagc ctgaggagcg gcccaccttc 2581 gagtacctgc aggccttcct ggaggactac ttcacgtcca ccgagcccca gttccagccc 2641 ggggagaacc tcctcgagga ctacaaggat gacgatgaca aggggcccta ttctatagtg 2701 tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc 2761 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct 2821 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 2881 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg 2941 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta 3001 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 3061 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 3121 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg 3181 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 3241 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 3301 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 3361 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 3421 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 3481 tccccaggca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg 3541 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 3601 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca 3661 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc 3721 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa 3781 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt 3841 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct 3901 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct 3961 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga 4021 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc 4081 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg 4141 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc 4201 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca 4261 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga 4321 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc 4381 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga 4441 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca 4501 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg 4561 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct 4621 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc 4681 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga 4741 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc 4801 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 4861 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 4921 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca 4981 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 5041 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 5101 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 5161 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 5221 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 5281 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 5341 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 5401 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 5461 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 5521 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa 5581 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 5641 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 5701 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 5761 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 5821 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 5881 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 5941 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 6001 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 6061 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 6121 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 6181 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 6241 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 6301 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 6361 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 6421 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 6481 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 6541 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 6601 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 6661 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 6721 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 6781 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 6841 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 6901 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 6961 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 7021 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 7081 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc //