LOCUS       sequence-236093-        5669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid containing a GST tag with 3 repeats of 
            the second SH3 domain from human NCK1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5669)
  AUTHORS   Li P, Banjade S, Cheng HC, Kim S, Chen B, Guo L, Llaguno M, 
            Hollingsworth JV, King DS, Banani SF, Russo PS, Jiang QX, Nixon BT, 
            Rosen MK
  TITLE     Phase transitions in the assembly of multivalent signalling 
            proteins.
  JOURNAL   Nature. 2012 Mar 7;483(7389):336-40. doi: 10.1038/nature10879.
  PUBMED    22398450
REFERENCE   2  (bases 1 to 5669)
  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"; journalName: 
            "Nature"; date: "2012-03-7- 7"; volume: "483"; issue: "7389"; pages:
            "336-40"
COMMENT     Sequence Label: pGEX SH3(2)-3R
FEATURES             Location/Qualifiers
     source          1..5669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     38..60
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             87..104
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             105..125
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(888..910)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1055..1073)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1141..1245
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1246..1314,1315..2106)
                     /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(1464..1483)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2277..2865
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2766..2785
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3019..3036
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3109..3186
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3187..4269
                     /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(3206..3225)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4318..4323,4324..4341,4342..4348)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4356..4372
                     /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     4380..4396
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4380..4396
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4392..4919
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4412..4429)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4412..4428)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4421..4443)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(5021..5026,5027..5042,5043..5049)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    5057..5073
                     /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             join(5096..5669,1..80)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
ORIGIN
        1 acttgaaatc cagcaagtat atagcatggc ctttgcaggg ctggcaagcc acgtttggtg
       61 gtggcgacca tcctccaaaa tcggatctgg ttccgcgtgg atctgaaaac ctgtattttc
      121 agggccatat ggacctcaac atgcccgctt atgtgaaatt taactacatg gctgagagag
      181 aggatgaatt atcattgata aaggggacaa aggtgatcgt catggagaaa agcagtgatg
      241 ggtggtggcg tggtagctac aatggacaag ttggatggtt cccttcaaac tatgtaactg
      301 aagaaggtga cagtcctttg gcgtctggtg cgggtggttc tgaaggtggt ggttctgaag
      361 gtggtacctc tggtgcgacc gacctcaaca tgcccgctta tgtgaaattt aactacatgg
      421 ctgagagaga ggatgaatta tcattgataa aggggacaaa ggtgatcgtc atggagaaaa
      481 gcagtgatgg gtggtggcgt ggtagctaca atggacaagt tggatggttc ccttcaaact
      541 atgtaactga agaaggtgac agtcctttgg catccggagc aggaggctcc gaaggaggag
      601 gctccgaggg aggaacatcc ggagcaacag acctcaacat gcccgcttat gtgaaattta
      661 actacatggc tgagagagag gatgaattat cattgataaa ggggacaaag gtgatcgtca
      721 tggagaaaag cagtgatggg tggtggcgtg gtagctacaa tggacaagtt ggatggttcc
      781 cttcaaacta tgtaactgaa gaaggtgaca gtcctttgtg aggatccgcg gccgcatcgt
      841 gactgactga cgatctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
      901 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
      961 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
     1021 gatagcggag tgtataattc ttgaagacga aagggcctcg tgatacgcct atttttatag
     1081 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     1141 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     1201 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     1261 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     1321 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     1381 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     1441 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     1501 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     1561 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     1621 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     1681 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     1741 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     1801 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     1861 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     1921 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     1981 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     2041 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     2101 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     2161 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     2221 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2281 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2341 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2401 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     2461 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2521 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2581 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2641 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2701 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2761 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2821 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2881 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2941 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3001 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     3061 tattttctcc ttacgcatct gtgcggtatt tcacaccgca taaattccga caccatcgaa
     3121 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
     3181 gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
     3241 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
     3301 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc
     3361 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa
     3421 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg
     3481 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc
     3541 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct
     3601 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt
     3661 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt
     3721 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg
     3781 gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc
     3841 gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt
     3901 cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc
     3961 gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac
     4021 agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa
     4081 accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg
     4141 ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct
     4201 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     4261 gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cccaggcttt
     4321 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     4381 aggaaacagc tatgaccatg attacggatt cactggccgt cgttttacaa cgtcgtgact
     4441 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     4501 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     4561 gcgaatggcg ctttgcctgg tttccggcac cagaagcggt gccggaaagc tggctggagt
     4621 gcgatcttcc tgaggccgat actgtcgtcg tcccctcaaa ctggcagatg cacggttacg
     4681 atgcgcccat ctacaccaac gtgacctatc ccattacggt caatccgccg tttgttccca
     4741 cggagaatcc gacgggttgt tactcgctca catttaatgt tgatgaaagc tggctacagg
     4801 aaggccagac gcgaattatt tttgatggcg ttggaattag cttatcgact gcacggtgca
     4861 ccaatgcttc tggcgtcagg cagccatcgg aagctgtggt atggctgtgc aggtcgtaaa
     4921 tcactgcata attcgtgtcg ctcaaggcgc actcccgttc tggataatgt tttttgcgcc
     4981 gacatcataa cggttctggc aaatattctg aaatgagctg ttgacaatta atcatcggct
     5041 cgtataatgt gtggaattgt gagcggataa caatttcaca caggaaacag tattcatgtc
     5101 ccctatacta ggttattgga aaattaaggg ccttgtgcaa cccactcgac ttcttttgga
     5161 atatcttgaa gaaaaatatg aagagcattt gtatgagcgc gatgaaggtg ataaatggcg
     5221 aaacaaaaag tttgaattgg gtttggagtt tcccaatctt ccttattata ttgatggtga
     5281 tgttaaatta acacagtcta tggccatcat acgttatata gctgacaagc acaacatgtt
     5341 gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt gaaggagcgg ttttggatat
     5401 tagatacggt gtttcgagaa ttgcatatag taaagacttt gaaactctca aagttgattt
     5461 tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat cgtttatgtc ataaaacata
     5521 tttaaatggt gatcatgtaa cccatcctga cttcatgttg tatgacgctc ttgatgttgt
     5581 tttatacatg gacccaatgt gcctggatgc gttcccaaaa ttagtttgtt ttaaaaaacg
     5641 tattgaagct atcccacaaa ttgataagt
//
