LOCUS       sequence-271602-        5800 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SH3BP1-NTD-GST fusion protein in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5800)
  AUTHORS   Elbediwy A, Zihni C, Terry SJ, Clark P, Matter K, Balda MS
  TITLE     Epithelial junction formation requires confinement of Cdc42 activity
            by a novel SH3BP1 complex.
  JOURNAL   J Cell Biol. 2012 Aug 20;198(4):677-93. doi: 10.1083/jcb.201202094. 
            Epub 2012 Aug 13.
  PUBMED    22891260
REFERENCE   2  (bases 1 to 5800)
  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.201202094"; journalName: "J Cell Biol"; date: 
            "2012-08-20- 20"; volume: "198"; issue: "4"; pages: "677-93"
COMMENT     Sequence Label: pGEX-4T-3-SH3BP1-NTD
FEATURES             Location/Qualifiers
     source          1..5800
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    8..24
                     /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             47..700
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     658..680
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             707..724
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1639..1661)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1806..1824)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1892..1996
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1997..2065,2066..2857)
                     /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(2215..2234)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3028..3616
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3517..3536
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3770..3787
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3860..3937
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3938..5020
                     /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(3957..3976)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5069..5074,5075..5092,5093..5099)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5107..5123
                     /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     5131..5147
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5131..5147
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5143..5670
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5163..5180)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5163..5179)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5172..5194)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(5772..5777,5778..5793,5794..5800)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 tgtggaattg tgagcggata acaatttcac acaggaaaca gtattcatgt cccctatact
       61 aggttattgg aaaattaagg gccttgtgca acccactcga cttcttttgg aatatcttga
      121 agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt gataaatggc gaaacaaaaa
      181 gtttgaattg ggtttggagt ttcccaatct tccttattat attgatggtg atgttaaatt
      241 aacacagtct atggccatca tacgttatat agctgacaag cacaacatgt tgggtggttg
      301 tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg gttttggata ttagatacgg
      361 tgtttcgaga attgcatata gtaaagactt tgaaactctc aaagttgatt ttcttagcaa
      421 gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt cataaaacat atttaaatgg
      481 tgatcatgta acccatcctg acttcatgtt gtatgacgct cttgatgttg ttttatacat
      541 ggacccaatg tgcctggatg cgttcccaaa attagtttgt tttaaaaaac gtattgaagc
      601 tatcccacaa attgataagt acttgaaatc cagcaagtat atagcatggc ctttgcaggg
      661 ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa tcggatctgg ttccgcgtgg
      721 atccatgcag gacctgccag gcaacgacaa cagcaccgcc ggcctcgaga tgatgaagag
      781 gcagctgcac cgcatgcggc agctggccca gacgggcagc ttgggacgca ccccggagac
      841 cgctgagttc ctgggtgagg acctgctgca ggtagaacag cggctggagc cggccaagcg
      901 ggcagcccac aacatccaca agcggctgca ggcctgtctg cagggccaga gcggggcaga
      961 catggacaag cgggtgaaga agcttcccct catggctctg tccaccacga tggctgagag
     1021 cttcaaggag ctggaccctg attccagcat ggggaaggcc ttggagatga gctgtgccat
     1081 ccagaatcag ctggcccgca tcctggccga gtttgagatg accctggaga gggacgtcct
     1141 gcagccactc agcaggctga gtgaggagga gctgccagcc atcctcaaac acaagaaaag
     1201 cctccagaag ctcgtgtccg actggaacac actcaagagc aggctcagtc aggcaaccaa
     1261 gaattcaggc agcagtcaag gcctaggagg cagcccgggt agtcacagcc atacgaccat
     1321 ggccaacaag gtggagacgc tgaaggagga ggaggaggag ctgaagagga aagtggagca
     1381 atgcagggac gagtacttgg ctgacctgta ccactttgtt accaaggagg actcctatgc
     1441 caactacttc attcgtctcc tggagattca ggccgattac catcgcaggt cactgagctc
     1501 gctggacaca gccctggctg agctgaggga gaaccacggc caagcagacc actccccttc
     1561 gtagtctagt cgactcgagc ggccgcatcg tgactgactg acgatctgcc tcgcgcgttt
     1621 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     1681 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     1741 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtataatt cttgaagacg
     1801 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     1861 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1921 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1981 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     2041 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     2101 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     2161 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     2221 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     2281 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2341 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     2401 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     2461 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     2521 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
     2581 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     2641 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     2701 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     2761 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     2821 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2881 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     2941 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3001 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3061 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3121 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     3181 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3241 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3301 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3361 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3421 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3481 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3541 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3601 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3661 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3721 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3781 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3841 ttcacaccgc ataaattccg acaccatcga atggcgcaaa acctttcgcg gtatggcatg
     3901 atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga
     3961 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
     4021 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
     4081 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
     4141 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
     4201 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
     4261 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     4321 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     4381 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
     4441 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     4501 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
     4561 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
     4621 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
     4681 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat
     4741 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac
     4801 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
     4861 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac
     4921 caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     4981 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa
     5041 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     5101 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacggat
     5161 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     5221 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     5281 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gctttgcctg gtttccggca
     5341 ccagaagcgg tgccggaaag ctggctggag tgcgatcttc ctgaggccga tactgtcgtc
     5401 gtcccctcaa actggcagat gcacggttac gatgcgccca tctacaccaa cgtgacctat
     5461 cccattacgg tcaatccgcc gtttgttccc acggagaatc cgacgggttg ttactcgctc
     5521 acatttaatg ttgatgaaag ctggctacag gaaggccaga cgcgaattat ttttgatggc
     5581 gttggaatta gcttatcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg
     5641 gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg
     5701 cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct
     5761 gaaatgagct gttgacaatt aatcatcggc tcgtataatg
//
