LOCUS       sequence-258859-        5797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GST::dCrk (based on crk isoform RA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5797)
  AUTHORS   Spracklen AJ, Thornton-Kolbe EM, Bonner AN, Florea A, Compton PJ, 
            Fernandez-Gonzalez R, Peifer M
  TITLE     The Crk adapter protein is essential for Drosophila embryogenesis, 
            where it regulates multiple actin-dependent morphogenic events.
  JOURNAL   Mol Biol Cell. 2019 Jul 18:mbcE19050302. doi: 
            10.1091/mbc.E19-05-0302.
  PUBMED    31318326
REFERENCE   2  (bases 1 to 5797)
  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"; journalName: "Mol Biol 
            Cell. 2019 Jul 18:mbcE19050302. doi: 10.1091/mbc.E19-05-0302."
COMMENT     Sequence Label: pGEX-6P-2_dCrk
FEATURES             Location/Qualifiers
     source          1..5797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..78
                     /gene="lacI"
                     /label=lacI promoter
     CDS             79..1161
                     /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(98..117)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1210..1215,1216..1233,1234..1240)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1248..1264
                     /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     1272..1288
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1272..1288
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1284..1811
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1304..1321)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1304..1320)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1313..1335)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1913..1918,1919..1934,1935..1941)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1949..1965
                     /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             1988..2641
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2599..2621
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2648..2671
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3577..3599)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3744..3762)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3830..3934
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3935..4003,4004..4795)
                     /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(4153..4172)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4966..5554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5455..5474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5708..5725
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
       61 caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt
      121 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
      181 cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa
      241 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac
      301 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg
      361 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt
      421 ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt
      481 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca
      541 cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg
      601 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg
      661 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg
      721 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
      781 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
      841 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
      901 gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
      961 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
     1021 ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccacgctagc gcccaatacg
     1081 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     1141 cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc
     1201 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     1261 acaatttcac acaggaaaca gctatgacca tgattacgga ttcactggcc gtcgttttac
     1321 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     1381 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     1441 gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc accagaagcg gtgccggaaa
     1501 gctggctgga gtgcgatctt cctgaggccg atactgtcgt cgtcccctca aactggcaga
     1561 tgcacggtta cgatgcgccc atctacacca acgtgaccta tcccattacg gtcaatccgc
     1621 cgtttgttcc cacggagaat ccgacgggtt gttactcgct cacatttaat gttgatgaaa
     1681 gctggctaca ggaaggccag acgcgaatta tttttgatgg cgttggaatt agcttatcga
     1741 ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg gtatggctgt
     1801 gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat
     1861 gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc tgttgacaat
     1921 taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     1981 agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc aacccactcg
     2041 acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg
     2101 tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc ttccttatta
     2161 tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata tagctgacaa
     2221 gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc
     2281 ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact ttgaaactct
     2341 caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag atcgtttatg
     2401 tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt tgtatgacgc
     2461 tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa aattagtttg
     2521 ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat ccagcaagta
     2581 tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc atcctccaaa
     2641 atcggatctg gaagttctgt tccaggggcc cctgggatcc atggatacat ttgacgtttc
     2701 tgataggaac agctggtact ttggtcccat gtctagacag gatgctactg aagttttgat
     2761 gaacgaacgc gagcggggag tgtttttagt ccgtgatagt aactcgatag caggggatta
     2821 tgtactttgt gtaagagaag atacaaaagt tagcaactac atcattaaca aagttcaaca
     2881 acaggatcaa atcgtttacc gcattgggga tcagtctttt gacaatctac cgaaactctt
     2941 aactttttac actcttcatt atttggatac aaccccttta aaacggcctg cgtgtagaag
     3001 ggtggaaaaa gtaataggaa agttcgattt cgttggcagc gatcaagatg atttaccttt
     3061 tcaaagaggt gaagttttaa caatagttcg aaaagacgag gatcaatggt ggactgcgcg
     3121 taactcctcg gggaaaattg gtcaaatacc ggttccctat atacaacagt atgacgatta
     3181 tatggatgaa gatgctattg ataaaaacga accttccatt tcgggatcta gcaatgtatt
     3241 tgaaagtact cttaaaagga cagatttaaa tcgaaaacta cctgcatacg cccgcgtaaa
     3301 acagtcaagg gtccctaacg catacgataa gactgcatta aaattggaaa taggtgacat
     3361 tattaaagtc actaaaacaa acattaatgg gcaatgggag ggagaattaa atggaaaaaa
     3421 tggtcatttt cccttcacgc acgttgaatt tgtcgatgat tgtgatttaa gcaaaaactc
     3481 cacagaaata tgctaagaat tcccgggtcg actcgagcgg ccgcatcgtg actgactgac
     3541 gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     3601 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     3661 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     3721 gtataattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     3781 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3841 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3901 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3961 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4021 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4081 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4141 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     4201 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4261 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4321 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4381 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4441 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     4501 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4561 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4621 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4681 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4741 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4801 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4861 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4921 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4981 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5041 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5101 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5161 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5221 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5281 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5341 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5401 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5461 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5521 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5581 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5641 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     5701 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     5761 tacgcatctg tgcggtattt cacaccgcat aaattcc
//
