LOCUS       sequence-236086-        5924 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Bacterial expression plasmid containing a GST tag with 4 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 5924)
  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 5924)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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)-4R
FEATURES             Location/Qualifiers
     source          1..5924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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             76..729
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     687..709
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             736..753
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             754..774
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1792..1814)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1959..1977)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2045..2149
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2150..2218,2219..3010)
                     /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(2368..2387)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3181..3769
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3670..3689
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3923..3940
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4013..4090
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4091..5173
                     /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(4110..4129)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5222..5227,5228..5245,5246..5252)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5260..5276
                     /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     5284..5300
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5284..5300
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5296..5823
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5316..5333)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5316..5332)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5325..5347)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa
      121 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc
      181 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt
      241 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata
      301 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt
      361 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt
      421 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat
      481 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg
      541 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa
      601 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc
      661 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat
      721 cctccaaaat cggatctggt tccgcgtgga tctgaaaacc tgtattttca gggccatatg
      781 gacctcaaca tgcccgctta tgtgaaattt aactacatgg ctgagagaga ggatgaatta
      841 tcattgataa aggggacaaa ggtgatcgtc atggagaaaa gcagtgatgg gtggtggcgt
      901 ggtagctaca atggacaagt tggatggttc ccttcaaact atgtaactga agaaggtgac
      961 agtcctttgg ccagcggcgc cggaggcagc gagggcggcg gaagcgaggg cggcacgtca
     1021 ggcgccacgc atatggacct caacatgccc gcttatgtga aatttaacta catggctgag
     1081 agagaggatg aattatcatt gataaagggg acaaaggtga tcgtcatgga gaaaagcagt
     1141 gatgggtggt ggcgtggtag ctacaatgga caagttggat ggttcccttc aaactatgta
     1201 actgaagaag gtgacagtcc tttggcgtct ggtgcgggtg gttctgaagg tggtggttct
     1261 gaaggtggta cctctggtgc gaccgacctc aacatgcccg cttatgtgaa atttaactac
     1321 atggctgaga gagaggatga attatcattg ataaagggga caaaggtgat cgtcatggag
     1381 aaaagcagtg atgggtggtg gcgtggtagc tacaatggac aagttggatg gttcccttca
     1441 aactatgtaa ctgaagaagg tgacagtcct ttggcatccg gagcaggagg ctccgaagga
     1501 ggaggctccg agggaggaac atccggagca acagacctca acatgcccgc ttatgtgaaa
     1561 tttaactaca tggctgagag agaggatgaa ttatcattga taaaggggac aaaggtgatc
     1621 gtcatggaga aaagcagtga tgggtggtgg cgtggtagct acaatggaca agttggatgg
     1681 ttcccttcaa actatgtaac tgaagaaggt gacagtcctt tgtgaggatc cgcggccgca
     1741 tcgtgactga ctgacgatct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     1801 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     1861 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
     1921 tagcgatagc ggagtgtata attcttgaag acgaaagggc ctcgtgatac gcctattttt
     1981 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     2041 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2101 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     2161 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     2221 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     2281 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     2341 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     2401 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     2461 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     2521 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     2581 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     2641 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     2701 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     2761 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     2821 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2881 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     2941 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3001 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     3061 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3121 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3181 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3241 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3301 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     3361 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3421 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3481 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3541 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3601 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3661 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3721 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3781 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3841 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3901 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     3961 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataaatt ccgacaccat
     4021 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
     4081 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
     4141 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
     4201 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
     4261 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
     4321 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
     4381 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
     4441 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
     4501 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
     4561 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
     4621 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
     4681 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
     4741 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
     4801 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
     4861 taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga
     4921 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
     4981 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
     5041 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
     5101 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     5161 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcaccccagg
     5221 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     5281 acacaggaaa cagctatgac catgattacg gattcactgg ccgtcgtttt acaacgtcgt
     5341 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     5401 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     5461 aatggcgaat ggcgctttgc ctggtttccg gcaccagaag cggtgccgga aagctggctg
     5521 gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct caaactggca gatgcacggt
     5581 tacgatgcgc ccatctacac caacgtgacc tatcccatta cggtcaatcc gccgtttgtt
     5641 cccacggaga atccgacggg ttgttactcg ctcacattta atgttgatga aagctggcta
     5701 caggaaggcc agacgcgaat tatttttgat ggcgttggaa ttagcttatc gactgcacgg
     5761 tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg
     5821 taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg
     5881 cgccgacatc ataacggttc tggcaaatat tctgaaatga gctg
//
