LOCUS       sequence-202048-        6911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            SORBS2-1/3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6911)
  AUTHORS   Teyra J, Huang H, Jain S, Guan X, Dong A, Liu Y, Tempel W, Min J, 
            Tong Y, Kim PM, Bader GD, Sidhu SS
  TITLE     Comprehensive Analysis of the Human SH3 Domain Family Reveals a Wide
            Variety of Non-canonical Specificities.
  JOURNAL   Structure. 2017 Oct 3;25(10):1598-1610.e3. doi: 
            10.1016/j.str.2017.07.017. Epub 2017 Sep 7.
  PUBMED    28890361
REFERENCE   2  (bases 1 to 6911)
  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: 
            "10.1016/j.str.2017.07.017"; journalName: "Structure"; date: 
            "2017-10-3- 3"; volume: "25"; issue: "10"; pages: "1598-1610.e3"
COMMENT     Sequence Label: pHH0103_SORBS2-1/3
FEATURES             Location/Qualifiers
     source          1..6911
                     /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)."
     primer_bind     42..64
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             138..155
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             171..824
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     782..804
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             831..854
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             867..884
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      1106..1153
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(2746..3201)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2883..2904)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3095..3114
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3243..3265)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3351..3491
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3406..3425
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3506..3523)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3677..4264)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4435..5226,5227..5295))
                     /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     5058..5077
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5296..5400)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5468..5486
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5509..5586
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             5587..6669
                     /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(5606..5625)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag ccagtccgtt taggtgtttt cacgagcact tcaccaacaa ggaccataga
      121 ttatgaaaat cgaagaacac catcaccatc accattccag cggtaagctt atgtccccta
      181 tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt ttggaatatc
      241 ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa tggcgaaaca
      301 aaaagtttga attgggtttg gagtttccca atcttcctta ttatattgat ggtgatgtta
      361 aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac atgttgggtg
      421 gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg gatattagat
      481 acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt gattttctta
      541 gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa acatatttaa
      601 atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat gttgttttat
      661 acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa aaacgtattg
      721 aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca tggcctttgc
      781 agggctggca agccacgttt ggtggtggcg accatcctcc aaaatcggat ctggaagttc
      841 tgttccaggg gcccctgtcc agcggtctgg ttccgcgtgg ttccggtacc gcggcccagc
      901 cggccatggc ccagggcgcg ctgctgccag caaaagccgt ttacgatttc aaagcccaga
      961 cctccaaaga actgtccttt aaaaaaggtg ataccgttta cattctgcgt aaaattgatc
     1021 agaactggta tgaaggtgaa catcatggtc gtgttggtat cttcccgatt tcctacgtcg
     1081 aaaactgacc gcggccgcat aataaaccga tacaattaaa ggctcctttt ggagcctttt
     1141 ttttggagat tttcaacgtg aaaaaattat tattcgcaat tcctttagtt gttcctttct
     1201 attctcactc cgctgaaact gttgaaagtt gtttagcaaa accccataca gaaaattcat
     1261 ttactaacgt ctggaaagac gacaaaactt tagatcgtta cgctaactat gagggctgtc
     1321 tgtggaatgc tacaggcgtt gtagtttgta ctggtgacga aactcagtgt ctagctagag
     1381 tggcggtggc tctggttccg gtgattttga ttatgaaaag atggcaaacg ctaataaggg
     1441 ggctatgacc gaaaatgccg atgaaaacgc gctacagtct gacgctaaag gcaaacttga
     1501 ttctgtcgct actgattacg gtgctgctat cgatggtttc attggtgacg tttccggcct
     1561 tgctaatggt aatggtgcta ctggtgattt tgctggctct aattcccaaa tggctcaagt
     1621 cggtgacggt gataattcac ctttaatgaa taatttccgt caatatttac cttccctccc
     1681 tcaatcggtt gaatgtcgcc cttttgtctt tggcgctggt aaaccatatg aattttctat
     1741 tgattgtgac aaaataaact tattccgtgg tgtctttgcg tttcttttat atgttgccac
     1801 ctttatgtat gtattttcta cgtttgctaa catactgcgt aataaggagt cttaatcatg
     1861 ccagttcttt tggctagcgc cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt
     1921 gcatggagcc gggccacctc gacctgaatg gaagccggcg gcacctcgct aacggattca
     1981 ccactccaag aattggagcc aatcaattct tgcggagaac tgtgaatgcg caaaccaacc
     2041 cttggcagaa catatccatc gcgtccgcca tctccagcag ccgcacgcgg cgcatctcgg
     2101 gcagcgttgg gtcctggcca cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc
     2161 taggctggcg gggttgcctt actggttagc agaatgaatc accgatacgc gagcgaacgt
     2221 gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg gtcttcggtt
     2281 tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga
     2341 tctgcatcgc aggatgctgc tggctaccct gtggaacacc tacatctgta ttaacgaagc
     2401 gctggcattg accctgagtg atttttctct ggtcccgccg catccatacc gccagttgtt
     2461 taccctcaca acgttccagt aaccgggcat gttcatcatc agtaacccgt atcgtgagca
     2521 tcctctctcg tttcatcggt atcattaccc ccatgaacag aaatccccct tacacggagg
     2581 catcagtgac caaacaggaa aaaaccgccc ttaacatggc ccgctttatc agaagccaga
     2641 cattaacgct tctggagaaa ctcaacgagc tggacgcgga tgaacaggca gacatctgtg
     2701 aatcgcttca cgaccacgct gatgagcttt accgcaggat ccgcgaaatt gtaagcgtta
     2761 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     2821 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     2881 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     2941 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3001 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3061 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3121 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3181 atgcgccgct acagggcgcg tcgcggatcc tgcctcgcgc gtttcggtga tgacggtgaa
     3241 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     3301 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
     3361 acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga
     3421 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     3481 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3541 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3601 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3661 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3721 gctcaagtca gaggtggcga aacccgacag gactataaag ataccagggt ttccccctgg
     3781 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3841 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3901 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3961 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4021 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4081 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     4141 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4201 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4261 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4321 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4381 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4441 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4501 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4561 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4621 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4681 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4741 tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4801 cggttcccaa cgatcaaggc gagttacatg atcccccata ttgtgcaaaa aagcggttag
     4861 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4921 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4981 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5041 cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5101 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5161 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5221 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5281 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5341 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5401 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     5461 ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaattcccga caccatcgaa
     5521 tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
     5581 gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
     5641 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
     5701 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc
     5761 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa
     5821 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg
     5881 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc
     5941 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct
     6001 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt
     6061 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt
     6121 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg
     6181 gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc
     6241 gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt
     6301 cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc
     6361 gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac
     6421 agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa
     6481 accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg
     6541 ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct
     6601 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     6661 gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac aattctcatg
     6721 tttgacagct tatcatcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg
     6781 gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg
     6841 cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct
     6901 gaaatgagct g
//
