LOCUS       sequence-251762-        6515 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GST-Src42a for purification from E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6515)
  AUTHORS   Williamson AP, Vale RD
  TITLE     Spatial control of Draper receptor signaling initiates apoptotic 
            cell engulfment.
  JOURNAL   J Cell Biol. 2018 Nov 5;217(11):3977-3992. doi: 
            10.1083/jcb.201711175. Epub 2018 Aug 23.
  PUBMED    30139739
REFERENCE   2  (bases 1 to 6515)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1083/jcb.201711175"; journalName: "J Cell Biol"; date: 
            "2018-11-5- 5"; volume: "217"; issue: "11"; pages: "3977-3992"
COMMENT     Sequence Label: pGEX Src42a
FEATURES             Location/Qualifiers
     source          1..6515
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      134..722
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     623..642
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     876..893
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        966..1043
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1044..2126
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1063..1082)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2175..2180,2181..2198,2199..2205)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2213..2229
                     /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)."
     primer_bind     2237..2253
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2237..2253
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2249..2776
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(2269..2286)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2269..2285)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2278..2300)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2878..2883,2884..2899,2900..2906)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2914..2930
                     /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)."
     CDS             2953..3606
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3564..3586
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             3613..3636
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(5260..5282)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5427..5445)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5513..5617
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5618..5686,5687..6478)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5836..5855)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
       61 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
      121 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
      181 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      241 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
      301 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      361 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      421 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      481 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      541 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      601 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      661 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      721 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      781 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      841 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      901 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcataa
      961 attccgacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
     1021 agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg
     1081 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
     1141 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
     1201 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
     1261 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
     1321 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
     1381 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
     1441 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
     1501 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
     1561 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
     1621 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
     1681 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
     1741 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
     1801 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
     1861 acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
     1921 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
     1981 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacg ctagcgccca
     2041 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     2101 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat
     2161 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     2221 ggataacaat ttcacacagg aaacagctat gaccatgatt acggattcac tggccgtcgt
     2281 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     2341 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     2401 gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt ccggcaccag aagcggtgcc
     2461 ggaaagctgg ctggagtgcg atcttcctga ggccgatact gtcgtcgtcc cctcaaactg
     2521 gcagatgcac ggttacgatg cgcccatcta caccaacgtg acctatccca ttacggtcaa
     2581 tccgccgttt gttcccacgg agaatccgac gggttgttac tcgctcacat ttaatgttga
     2641 tgaaagctgg ctacaggaag gccagacgcg aattattttt gatggcgttg gaattagctt
     2701 atcgactgca cggtgcacca atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg
     2761 gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg
     2821 ataatgtttt ttgcgccgac atcataacgg ttctggcaaa tattctgaaa tgagctgttg
     2881 acaattaatc atcggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag
     2941 gaaacagtat tcatgtcccc tatactaggt tattggaaaa ttaagggcct tgtgcaaccc
     3001 actcgacttc ttttggaata tcttgaagaa aaatatgaag agcatttgta tgagcgcgat
     3061 gaaggtgata aatggcgaaa caaaaagttt gaattgggtt tggagtttcc caatcttcct
     3121 tattatattg atggtgatgt taaattaaca cagtctatgg ccatcatacg ttatatagct
     3181 gacaagcaca acatgttggg tggttgtcca aaagagcgtg cagagatttc aatgcttgaa
     3241 ggagcggttt tggatattag atacggtgtt tcgagaattg catatagtaa agactttgaa
     3301 actctcaaag ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt cgaagatcgt
     3361 ttatgtcata aaacatattt aaatggtgat catgtaaccc atcctgactt catgttgtat
     3421 gacgctcttg atgttgtttt atacatggac ccaatgtgcc tggatgcgtt cccaaaatta
     3481 gtttgtttta aaaaacgtat tgaagctatc ccacaaattg ataagtactt gaaatccagc
     3541 aagtatatag catggccttt gcagggctgg caagccacgt ttggtggtgg cgaccatcct
     3601 ccaaaatcgg atctggaagt tctgttccag gggcccctgg gatccatggg taactgcctc
     3661 accacacaga agggcgaacc cgacaagccc gcagatcgaa tcaagctgga cgacccgccc
     3721 accatcggag tcggagtggg cgtgccacaa atccccatgc cctcacacgc cggacagcca
     3781 ccggagcaga tacgtccggt tccccagatc ccggagagcg aaacggcagg tgccaacgcc
     3841 aagatttttg tcgccctcta cgactacgac gcccgcaccg acgaggattt gagcttccgc
     3901 aagggagagc acttggagat actgaatgac acgcagggtg actggtggct ggcgcggagc
     3961 aagaagacac gttcggaagg ctacattcca tccaattatg tggccaagtt gaaatcaatc
     4021 gaagcagaac cctggtactt ccgcaaaatc aaacgcattg aggctgagaa aaaacttcta
     4081 ctgccagaga acgagcacgg tgcattttta attcgcgatt ccgaaagccg tcacaacgac
     4141 tactcgctat cagtgcgcga tggcgatacg gttaagcatt atcgcatcag acaattggac
     4201 gaaggcggct tcttcatcgc caggcgcacg acattcagaa cccttcagga gctggtggaa
     4261 cactattcga aggactctga tggcctatgc gtcaacctct gcaagccgtg tgtccagatc
     4321 gagaagcctg taactgaggg gctttcgcac cgcactcgcg atcagtggga gatcgacaga
     4381 acatccttga aattcgtgcg caaactgggc tccggacagt ttggcgatgt ctgggaggga
     4441 ttgtggaaca acacaacacc tgtggcaatt aaaactctga aatctggcac aatggacccc
     4501 aaggatttct tagcggaagc ccagatcatg aagaaactgc gccacaccaa gcttatacag
     4561 ttgtacgctg tctgcactgt tgaggagcct atctatatta tcacagagtt aatgaagcac
     4621 ggttcactgt tggaatatct ccaagccatt gcaggcaagg gtcgtagcct taaaatgcaa
     4681 actctgattg atatggcagc gcaaatagct gctggcatgg cttacttgga gtcccagaat
     4741 tatattcata gggatttagc ggcgcgcaat gtactggtag gcgatggaaa catcgtcaaa
     4801 atcgccgact ttggtttagc taggctcatc aaggaggacg aatacgaggc gcgggtaggc
     4861 gccagatttc ccataaaatg gaccgctcca gaggctgcta actacagcaa attctcaata
     4921 aaatcggatg tttggagctt tggcattctt ctcacagaac tggtcaccta cggacgcata
     4981 ccatatccag gcatgaccaa cgctgaggtg ctaacgcaag tggagcacgg ctatcgaatg
     5041 ccgcaacctc ccaactgcga gccgcgcctg tatgagatta tgctggaatg ttggcacaag
     5101 gaccccatgc gcagacccac gtttgagacg ctacaatgga aactggaaga cttctataca
     5161 tctgatcaga gcgactacaa agaggcgcag gcctactgag cggccgcatc gtgactgact
     5221 gacgatctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     5281 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     5341 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
     5401 agtgtataat tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     5461 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     5521 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5581 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5641 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5701 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5761 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5821 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     5881 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5941 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6001 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6061 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6121 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     6181 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6241 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6301 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6361 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6421 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6481 gtcagaccaa gtttactcat atatacttta gattg
//
