LOCUS       sequence-271747-        7114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SHBP1-GST fusion protein in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7114)
  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 7114)
  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
FEATURES             Location/Qualifiers
     source          1..7114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..654
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     612..634
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             661..678
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2907..2929)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3074..3092)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3160..3264
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3265..3333,3334..4125)
                     /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(3483..3502)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4296..4884
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4785..4804
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5038..5055
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5128..5205
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5206..6288
                     /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(5225..5244)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6337..6342,6343..6360,6361..6367)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6375..6391
                     /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     6399..6415
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6399..6415
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             6411..6938
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6431..6448)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6431..6447)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6440..6462)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(7040..7045,7046..7061,7062..7068)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    7076..7092
                     /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)."
ORIGIN
        1 atgtccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt
       61 ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa
      121 tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta ttatattgat
      181 ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac
      241 atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg
      301 gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt
      361 gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa
      421 acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat
      481 gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa
      541 aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca
      601 tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc aaaatcggat
      661 ctggttccgc gtggatccat gcaggacctg ccaggcaacg acaacagcac cgccggcctc
      721 gagatgatga agaggcagct gcaccgcatg cggcagctgg cccagacggg cagcttggga
      781 cgcaccccgg agaccgctga gttcctgggt gaggacctgc tgcaggtaga acagcggctg
      841 gagccggcca agcgggcagc ccacaacatc cacaagcggc tgcaggcctg tctgcagggc
      901 cagagcgggg cagacatgga caagcgggtg aagaagcttc ccctcatggc tctgtccacc
      961 acgatggctg agagcttcaa ggagctggac cctgattcca gcatggggaa ggccttggag
     1021 atgagctgtg ccatccagaa tcagctggcc cgcatcctgg ccgagtttga gatgaccctg
     1081 gagagggacg tcctgcagcc actcagcagg ctgagtgagg aggagctgcc agccatcctc
     1141 aaacacaaga aaagcctcca gaagctcgtg tccgactgga acacactcaa gagcaggctc
     1201 agtcaggcaa ccaagaattc aggcagcagt caaggcctag gaggcagccc gggtagtcac
     1261 agccatacga ccatggccaa caaggtggag acgctgaagg aggaggagga ggagctgaag
     1321 aggaaagtgg agcaatgcag ggacgagtac ttggctgacc tgtaccactt tgttaccaag
     1381 gaggactcct atgccaacta cttcattcgt ctcctggaga ttcaggccga ttaccatcgc
     1441 aggtcactga gctcgctgga cacagccctg gctgagctga gggagaacca cggccaagca
     1501 gaccactccc cttcgatgac agccacccac ttccccaggg tgtatggggt gtcgctggca
     1561 acccacctgc aagagctggg ccgggagatt gccctgccca tcgaggcctg cgtcatgatg
     1621 ctgctttctg agggcatgaa ggaagagggt ctcttccgtc tggctgctgg ggcctcggtg
     1681 ctgaagcgtc tcaagcagac aatggcctcg gacccccaca gcctggagga gttctgctcc
     1741 gacccgcacg ctgtggcagg tgccctcaag tcctatctgc gggagctgcc agagcctctg
     1801 atgaccttcg acctctatga tgactggatg agggcagcca gcctgaagga gccaggggcc
     1861 cggctgcagg ccctccaaga ggtgtgcagc cgcctacccc ccgagaacct cagcaacctc
     1921 aggtacctga tgaagttcct ggcacggctg gccgaggagc aggaggtgaa caagatgaca
     1981 cccagcaaca tcgccatagt cctgggaccc aacttgctgt ggccacctga gaaagaaggg
     2041 gaccaggccc agctggatgc agcctccgtg tcttccatcc aggtggtggg cgtcgtcgag
     2101 gcgctgatcc agagcgcaga caccctcttc cctggagaca tcaacttcaa cgtgtcaggc
     2161 ctcttctcag ctgttaccct ccaggacaca gtcagtgaca ggctggcctc tgaggaactt
     2221 ccgtccactg ccgtgcccac cccagccacc accccggctc cggctccggc tccagctcca
     2281 gctccggccc cagccttggc ttcagcagct accaaggaaa ggacagagtc tgaggtgcct
     2341 cccagaccag cctcccccaa ggtcaccagg agtcccccgg agacagctgc cccagtggag
     2401 gacatggctc ggaggaccaa gcgcccggcg ccagcccggc ccaccatgcc gcccccccag
     2461 gtctccggct cccgctcctc ccctccagcc ccgcccttgc cccctggctc tggcagccct
     2521 gggacccccc aagccctgcc ccgacgtctg gttggcagca gcctccgagc ccccacagtg
     2581 ccacccccgt taccccccac accccctcag cctgcccggc gccaaagccg gcgttcacca
     2641 gcctccccca gcccggcctc cccaggtcca gcctccccca gcccagtctc tttgagtaac
     2701 cctgcacagg tggacctggg ggctgccaca gcagagggag gagcccctga ggctatcagt
     2761 ggggtcccca ctcccccagc tatccccccg cagccccgcc ccaggagcct tgcctcagag
     2821 accaactgat ctaggggtcg actcgagcgg ccgcatcgtg actgactgac gatctgcctc
     2881 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     2941 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     3001 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtataattct
     3061 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     3121 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3181 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3241 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3301 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3361 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3421 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     3481 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     3541 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3601 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3661 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3721 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3781 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     3841 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     3901 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     3961 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     4021 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4081 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4141 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4201 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4261 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4321 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4381 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4441 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4501 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4561 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4621 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4681 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4741 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4801 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4861 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4921 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4981 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     5041 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     5101 tgcggtattt cacaccgcat aaattccgac accatcgaat ggcgcaaaac ctttcgcggt
     5161 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg
     5221 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
     5281 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
     5341 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     5401 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     5461 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     5521 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     5581 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     5641 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     5701 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     5761 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     5821 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     5881 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     5941 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt
     6001 gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg
     6061 ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg
     6121 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa
     6181 agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     6241 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     6301 taatgtaagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     6361 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     6421 ttacggattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     6481 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     6541 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc tttgcctggt
     6601 ttccggcacc agaagcggtg ccggaaagct ggctggagtg cgatcttcct gaggccgata
     6661 ctgtcgtcgt cccctcaaac tggcagatgc acggttacga tgcgcccatc tacaccaacg
     6721 tgacctatcc cattacggtc aatccgccgt ttgttcccac ggagaatccg acgggttgtt
     6781 actcgctcac atttaatgtt gatgaaagct ggctacagga aggccagacg cgaattattt
     6841 ttgatggcgt tggaattagc ttatcgactg cacggtgcac caatgcttct ggcgtcaggc
     6901 agccatcgga agctgtggta tggctgtgca ggtcgtaaat cactgcataa ttcgtgtcgc
     6961 tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg acatcataac ggttctggca
     7021 aatattctga aatgagctgt tgacaattaa tcatcggctc gtataatgtg tggaattgtg
     7081 agcggataac aatttcacac aggaaacagt attc
//
