LOCUS Exported 7181 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses HA-tagged human Src-Y419F in mammalian cells for structure-function studies. ACCESSION . VERSION . KEYWORDS pDEST40-2XHA-Src-Y419F SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7181) 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 7181) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7181 /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(2562..2586) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2597..2623 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2627..2653 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2679..2720 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 2730..2747 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(2770..2787) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2776..3000 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3046..3474 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3133..3152) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3343..3364 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3483..3503) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3488..3817 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3668..3803 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3730..3749 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3884..4678 /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(3938..3957) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4548..4567 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4854..4975 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4891..4910) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4945..4964 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5024..5040) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5024..5040) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5037..5059) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5048..5064 /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(5072..5102) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5117..5138 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5255..5272) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5426..6014) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5506..5525) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6185..7045) /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 6808..6827 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7046..7150) /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 agcctggagc ccgccgagaa 1021 cgtgcacggc gctggcgggg gcgctttccc cgcctcgcag acccccagca agccagcctc 1081 ggccgacggc caccgcggcc ccagcgcggc cttcgccccc gcggccgccg agcccaagct 1141 gttcggaggc ttcaactcct cggacaccgt cacctccccg cagagggcgg gcccgctggc 1201 cggtggagtg accacctttg tggccctcta tgactatgag tctaggacgg agacagacct 1261 gtccttcaag aaaggcgagc ggctccagat tgtcaacaac acagagggag actggtggct 1321 ggcccactcg ctcagcacag gacagacagg ctacatcccc agcaactacg tggcgccctc 1381 cgactccatc caggctgagg agtggtattt tggcaagatc accagacggg agtcagagcg 1441 gttactgctc aatgcagaga acccgagagg gaccttcctc gtgcgagaaa gtgagaccac 1501 gaaaggtgcc tactgcctct cagtgtctga cttcgacaac gccaagggcc tcaacgtgaa 1561 gcactacaag atccgcaagc tggacagcgg cggcttctac atcacctccc gcacccagtt 1621 caacagcctg cagcagctgg tggcctacta ctccaaacac gccgatggcc tgtgccaccg 1681 cctcaccacc gtgtgcccca cgtccaagcc gcagactcag ggcctggcca aggatgcctg 1741 ggagatccct cgggagtcgc tgcggctgga ggtcaagctg ggccagggct gctttggcga 1801 ggtgtggatg gggacctgga acggtaccac cagggtggcc atcaaaaccc tgaagcctgg 1861 cacgatgtct ccagaggcct tcctgcagga ggcccaggtc atgaagaagc tgaggcatga 1921 gaagctggtg cagttgtatg ctgtggtttc agaggagccc atttacatcg tcacggagta 1981 catgagcaag gggagtttgc tggactttct caagggggag acaggcaagt acctgcggct 2041 gcctcagctg gtggacatgg ctgctcagat cgcctcaggc atggcgtacg tggagcggat 2101 gaactacgtc caccgggacc ttcgtgcagc caacatcctg gtgggagaga acctggtgtg 2161 caaagtggcc gactttgggc tggctcggct cattgaagac aatgagttca cggcgcggca 2221 aggtgccaaa ttccccatca agtggacggc tccagaagct gccctctatg gccgcttcac 2281 catcaagtcg gacgtgtggt ccttcgggat cctgctgact gagctcacca caaagggacg 2341 ggtgccctac cctgggatgg tgaaccgcga ggtgctggac caggtggagc ggggctaccg 2401 gatgccctgc ccgccggagt gtcccgagtc cctgcacgac ctcatgtgcc agtgctggcg 2461 gaaggagcct gaggagcggc ccaccttcga gtacctgcag gccttcctgg aggactactt 2521 cacgtccacc gagccccagt accagcccgg ggagaacctc gacccagctt tcttgtacaa 2581 agtggttgat ctagagtacc catacgatgt tccagattac gctggatacc catacgatgt 2641 tccagattac gcttgatcta gagggcccgc ggttcgaagg taagcctatc cctaaccctc 2701 tcctcggtct cgattctacg cgtaccggtc atcatcacca tcaccattga gtttaaaccc 2761 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg 2821 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa 2881 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca 2941 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg 3001 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg 3061 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg 3121 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc 3181 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc 3241 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga 3301 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa 3361 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga 3421 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct 3481 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 3541 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc 3601 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac 3661 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact 3721 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta 3781 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc 3841 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg 3901 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca 3961 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt 4021 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg 4081 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga 4141 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca 4201 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct 4261 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac 4321 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc 4381 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt 4441 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt 4501 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg 4561 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat 4621 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc 4681 gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt 4741 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg 4801 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt 4861 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag 4921 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg 4981 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg 5041 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 5101 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 5161 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 5221 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 5281 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 5341 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 5401 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 5461 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 5521 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 5581 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 5641 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 5701 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 5761 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 5821 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 5881 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 5941 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 6001 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 6061 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 6121 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 6181 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 6241 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 6301 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 6361 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 6421 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 6481 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 6541 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 6601 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 6661 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 6721 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 6781 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 6841 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 6901 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 6961 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 7021 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 7081 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 7141 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c //