LOCUS Exported 6320 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses Flag-tagged human Src-S4-UD-S32 in mammalian cells for structure-function studies. ACCESSION . VERSION . KEYWORDS pDEST40-2XFL-Src-S4-UD-S32 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6320) 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 6320) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6320 /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(1707..1731) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 1742..1765 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 1769..1792 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 1818..1859 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 1869..1886 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(1909..1926) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1915..2139 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2185..2613 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2272..2291) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2482..2503 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2622..2642) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2627..2956 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2807..2942 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2869..2888 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3023..3817 /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(3077..3096) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3687..3706 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3993..4114 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4030..4049) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4084..4103 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4163..4179) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4163..4179) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4176..4198) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4187..4203 /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(4211..4241) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4256..4277 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4394..4411) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4565..5153) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4645..4664) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5324..6184) /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 5947..5966 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6185..6289) /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 cgtccgaccc agctttcttg tacaaagtgg ttgatctaga 1741 ggattacaag gatgacgacg ataagggaga ttacaaggat gacgacgata agtgatctag 1801 agggcccgcg gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc 1861 gtaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc 1921 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag 1981 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta 2041 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag 2101 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca 2161 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg 2221 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg 2281 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg 2341 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt 2401 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt 2461 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta 2521 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa 2581 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg 2641 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta 2701 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 2761 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac 2821 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga 2881 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg 2941 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga 3001 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc 3061 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga 3121 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct 3181 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac 3241 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct 3301 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt 3361 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt 3421 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt 3481 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag 3541 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt 3601 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg 3661 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg 3721 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg 3781 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa 3841 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct 3901 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 3961 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt 4021 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 4081 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta 4141 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 4201 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 4261 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 4321 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 4381 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 4441 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 4501 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 4561 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 4621 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 4681 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 4741 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 4801 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 4861 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 4921 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 4981 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 5041 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 5101 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 5161 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 5221 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 5281 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 5341 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 5401 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 5461 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 5521 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 5581 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 5641 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 5701 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 5761 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 5821 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 5881 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 5941 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 6001 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 6061 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 6121 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 6181 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 6241 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 6301 gaaaagtgcc acctgacgtc //