LOCUS Exported 6473 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses Flag-tagged human Src-S4-KD in mammalian cells for structure-function studies. ACCESSION . VERSION . KEYWORDS pDEST40-2XFL-Src-S4-KD SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6473) AUTHORS Spassov DS, Ruiz-Saenz A, Piple A, Moasser MM TITLE A Dimerization Function in the Intrinsically Disordered N-Terminal Region of Src. JOURNAL Cell Rep. 2018 Oct 9;25(2):449-463.e4. doi: 10.1016/j.celrep.2018.09.035. PUBMED 30304684 REFERENCE 2 (bases 1 to 6473) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6473 /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" protein_bind 911..935 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(1860..1884) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 1895..1918 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 1922..1945 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 1971..2012 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 2022..2039 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(2062..2079) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2068..2292 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2338..2766 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2425..2444) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2635..2656 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2775..2795) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2780..3109 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2960..3095 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3022..3041 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3176..3970 /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(3230..3249) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3840..3859 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4146..4267 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4183..4202) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4237..4256 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4316..4332) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4316..4332) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4329..4351) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4340..4356 /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(4364..4394) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4409..4430 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4547..4564) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4718..5306) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4798..4817) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5477..6337) /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 6100..6119 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6338..6442) /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 gctggctagt 901 taagctatca acaagtttgt acaaaaaagc aggcttcgaa ggagatagaa ccatgggtag 961 caacaagagc aagcccaagg atgccagcca gcggcgccgc agcaagccgc agactcaggg 1021 cctggccaag gatgcctggg agatccctcg ggagtcgctg cggctggagg tcaagctggg 1081 ccagggctgc tttggcgagg tgtggatggg gacctggaac ggtaccacca gggtggccat 1141 caaaaccctg aagcctggca cgatgtctcc agaggccttc ctgcaggagg cccaggtcat 1201 gaagaagctg aggcatgaga agctggtgca gttgtatgct gtggtttcag aggagcccat 1261 ttacatcgtc acggagtaca tgagcaaggg gagtttgctg gactttctca agggggagac 1321 aggcaagtac ctgcggctgc ctcagctggt ggacatggct gctcagatcg cctcaggcat 1381 ggcgtacgtg gagcggatga actacgtcca ccgggacctt cgtgcagcca acatcctggt 1441 gggagagaac ctggtgtgca aagtggccga ctttgggctg gctcggctca ttgaagacaa 1501 tgagtacacg gcgcggcaag gtgccaaatt ccccatcaag tggacggctc cagaagctgc 1561 cctctatggc cgcttcacca tcaagtcgga cgtgtggtcc ttcgggatcc tgctgactga 1621 gctcaccaca aagggacggg tgccctaccc tgggatggtg aaccgcgagg tgctggacca 1681 ggtggagcgg ggctaccgga tgccctgccc gccggagtgt cccgagtccc tgcacgacct 1741 catgtgccag tgctggcgga aggagcctga ggagcggccc accttcgagt acctgcaggc 1801 cttcctggag gactacttca cgtccaccga gccccagtac cagcccgggg agaacctcga 1861 cccagctttc ttgtacaaag tggttgatct agaggattac aaggatgacg acgataaggg 1921 agattacaag gatgacgacg ataagtgatc tagagggccc gcggttcgaa ggtaagccta 1981 tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg tcatcatcac catcaccatt 2041 gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt 2101 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat 2161 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg 2221 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg 2281 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg 2341 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta 2401 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt 2461 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg 2521 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat 2581 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac 2641 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag 2701 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg 2761 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc 2821 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc 2881 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt 2941 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc 3001 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct 3061 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg 3121 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat 3181 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta 3241 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca 3301 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga 3361 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga 3421 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct 3481 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg 3541 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga 3601 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca 3661 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga 3721 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg 3781 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc 3841 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt 3901 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga 3961 gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc 4021 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt 4081 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc 4141 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 4201 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 4261 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat 4321 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa 4381 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc 4441 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc 4501 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact 4561 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac 4621 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa 4681 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg 4741 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa 4801 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc 4861 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac 4921 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac 4981 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg 5041 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt 5101 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa 5161 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct 5221 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga 5281 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 5341 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 5401 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 5461 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 5521 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg 5581 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag 5641 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt 5701 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag 5761 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt 5821 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca 5881 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg 5941 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat 6001 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta 6061 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca 6121 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct 6181 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat 6241 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa 6301 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt 6361 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa 6421 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc //