LOCUS       sequence-271744-        5620 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SH3BP1-CTD-GST fusion protein in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5620)
  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 5620)
  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-CTD
FEATURES             Location/Qualifiers
     source          1..5620
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             23..676
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     634..656
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             683..700
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1435..1457)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1602..1620)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1688..1792
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1793..1861,1862..2653)
                     /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(2011..2030)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2824..3412
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3313..3332
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3566..3583
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3656..3733
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3734..4816
                     /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(3753..3772)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4865..4870,4871..4888,4889..4895)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4903..4919
                     /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     4927..4943
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4927..4943
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4939..5466
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4959..4976)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4959..4975)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4968..4990)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(5568..5573,5574..5589,5590..5596)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    5604..5620
                     /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 tttcacacag gaaacagtat tcatgtcccc tatactaggt tattggaaaa ttaagggcct
       61 tgtgcaaccc actcgacttc ttttggaata tcttgaagaa aaatatgaag agcatttgta
      121 tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt gaattgggtt tggagtttcc
      181 caatcttcct tattatattg atggtgatgt taaattaaca cagtctatgg ccatcatacg
      241 ttatatagct gacaagcaca acatgttggg tggttgtcca aaagagcgtg cagagatttc
      301 aatgcttgaa ggagcggttt tggatattag atacggtgtt tcgagaattg catatagtaa
      361 agactttgaa actctcaaag ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt
      421 cgaagatcgt ttatgtcata aaacatattt aaatggtgat catgtaaccc atcctgactt
      481 catgttgtat gacgctcttg atgttgtttt atacatggac ccaatgtgcc tggatgcgtt
      541 cccaaaatta gtttgtttta aaaaacgtat tgaagctatc ccacaaattg ataagtactt
      601 gaaatccagc aagtatatag catggccttt gcagggctgg caagccacgt ttggtggtgg
      661 cgaccatcct ccaaaatcgg atctggttcc gcgtggatcc atgcaggacc tgccaggcaa
      721 cgacaacagc accgccggcc tcgaggaact tccgtccact gccgtgccca ccccagccac
      781 caccccggct ccggctccgg ctccagctcc agctccggcc ccagccttgg cttcagcagc
      841 taccaaggaa aggacagagt ctgaggtgcc tcccagacca gcctccccca aggtcaccag
      901 gagtcccccg gagacagctg ccccagtgga ggacatggct cggaggacca agcgcccggc
      961 gccagcccgg cccaccatgc cgccccccca ggtctccggc tcccgctcct cccctccagc
     1021 cccgcccttg ccccctggct ctggcagccc tgggaccccc caagccctgc cccgacgtct
     1081 ggttggcagc agcctccgag cccccacagt gccacccccg ttacccccca caccccctca
     1141 gcctgcccgg cgccaaagcc ggcgttcacc agcctccccc agcccggcct ccccaggtcc
     1201 agcctccccc agcccagtct ctttgagtaa ccctgcacag gtggacctgg gggctgccac
     1261 agcagaggga ggagcccctg aggctatcag tggggtcccc actcccccag ctatcccccc
     1321 gcagccccgc cccaggagcc ttgcctcaga gaccaactga tctagtcgac tcgagcggcc
     1381 gcatcgtgac tgactgacga tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg
     1441 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     1501 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc
     1561 acgtagcgat agcggagtgt ataattcttg aagacgaaag ggcctcgtga tacgcctatt
     1621 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     1681 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     1741 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1801 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1861 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1921 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1981 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     2041 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     2101 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     2161 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     2221 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     2281 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc
     2341 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     2401 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     2461 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     2521 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     2581 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     2641 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     2701 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2761 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2821 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2881 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2941 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     3001 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3061 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3121 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3181 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3241 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3301 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3361 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3421 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3481 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3541 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     3601 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcataa attccgacac
     3661 catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt
     3721 cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc
     3781 ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga
     3841 aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact
     3901 ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc
     3961 gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt
     4021 gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc
     4081 gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt
     4141 ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat
     4201 caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc
     4261 attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct
     4321 gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg
     4381 ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg
     4441 catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc
     4501 cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac
     4561 cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct
     4621 ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa
     4681 tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac
     4741 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     4801 ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcacccc
     4861 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     4921 ttcacacagg aaacagctat gaccatgatt acggattcac tggccgtcgt tttacaacgt
     4981 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     5041 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     5101 ctgaatggcg aatggcgctt tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg
     5161 ctggagtgcg atcttcctga ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac
     5221 ggttacgatg cgcccatcta caccaacgtg acctatccca ttacggtcaa tccgccgttt
     5281 gttcccacgg agaatccgac gggttgttac tcgctcacat ttaatgttga tgaaagctgg
     5341 ctacaggaag gccagacgcg aattattttt gatggcgttg gaattagctt atcgactgca
     5401 cggtgcacca atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg gctgtgcagg
     5461 tcgtaaatca ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg ataatgtttt
     5521 ttgcgccgac atcataacgg ttctggcaaa tattctgaaa tgagctgttg acaattaatc
     5581 atcggctcgt ataatgtgtg gaattgtgag cggataacaa
//
