LOCUS       sequence-191003-        6906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            FCHSD1-2/2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6906)
  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 6906)
  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_FCHSD1-2/2
FEATURES             Location/Qualifiers
     source          1..6906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..18
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             34..687
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     645..667
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             694..717
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             730..747
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      964..1011
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(2604..3059)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2741..2762)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2953..2972
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3101..3123)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3209..3349
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3264..3283
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3364..3381)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3535..4122)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4293..5084,5085..5153))
                     /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     4916..4935
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5154..5258)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5326..5344
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5367..5444
                     /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             5445..6527
                     /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(5464..5483)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6770..6775,6776..6791,6792..6798)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    6806..6822
                     /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     6811..6833
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caccatcacc atcaccattc cagcggtaag cttatgtccc ctatactagg ttattggaaa
       61 attaagggcc ttgtgcaacc cactcgactt cttttggaat atcttgaaga aaaatatgaa
      121 gagcatttgt atgagcgcga tgaaggtgat aaatggcgaa acaaaaagtt tgaattgggt
      181 ttggagtttc ccaatcttcc ttattatatt gatggtgatg ttaaattaac acagtctatg
      241 gccatcatac gttatatagc tgacaagcac aacatgttgg gtggttgtcc aaaagagcgt
      301 gcagagattt caatgcttga aggagcggtt ttggatatta gatacggtgt ttcgagaatt
      361 gcatatagta aagactttga aactctcaaa gttgattttc ttagcaagct acctgaaatg
      421 ctgaaaatgt tcgaagatcg tttatgtcat aaaacatatt taaatggtga tcatgtaacc
      481 catcctgact tcatgttgta tgacgctctt gatgttgttt tatacatgga cccaatgtgc
      541 ctggatgcgt tcccaaaatt agtttgtttt aaaaaacgta ttgaagctat cccacaaatt
      601 gataagtact tgaaatccag caagtatata gcatggcctt tgcagggctg gcaagccacg
      661 tttggtggtg gcgaccatcc tccaaaatcg gatctggaag ttctgttcca ggggcccctg
      721 tccagcggtc tggttccgcg tggttccggt accgcggccc agccggcctt tctggctcaa
      781 gctctgtact cctatacggg tcaaagcgct gaagaactgt cgtttccgga aggtgccctg
      841 attcgcctgc tgccgcgtgc gcaggatggc gtcgatgacg gtttttggcg tggtgaattc
      901 ggcggtcgcg tgggtgtttt tccgagcctg ctggttgaag aactgctggc ggccgcataa
      961 taaaccgata caattaaagg ctccttttgg agcctttttt ttggagattt tcaacgtgaa
     1021 aaaattatta ttcgcaattc ctttagttgt tcctttctat tctcactccg ctgaaactgt
     1081 tgaaagttgt ttagcaaaac cccatacaga aaattcattt actaacgtct ggaaagacga
     1141 caaaacttta gatcgttacg ctaactatga gggctgtctg tggaatgcta caggcgttgt
     1201 agtttgtact ggtgacgaaa ctcagtgtct agctagagtg gcggtggctc tggttccggt
     1261 gattttgatt atgaaaagat ggcaaacgct aataaggggg ctatgaccga aaatgccgat
     1321 gaaaacgcgc tacagtctga cgctaaaggc aaacttgatt ctgtcgctac tgattacggt
     1381 gctgctatcg atggtttcat tggtgacgtt tccggccttg ctaatggtaa tggtgctact
     1441 ggtgattttg ctggctctaa ttcccaaatg gctcaagtcg gtgacggtga taattcacct
     1501 ttaatgaata atttccgtca atatttacct tccctccctc aatcggttga atgtcgccct
     1561 tttgtctttg gcgctggtaa accatatgaa ttttctattg attgtgacaa aataaactta
     1621 ttccgtggtg tctttgcgtt tcttttatat gttgccacct ttatgtatgt attttctacg
     1681 tttgctaaca tactgcgtaa taaggagtct taatcatgcc agttcttttg gctagcgccg
     1741 ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga
     1801 cctgaatgga agccggcggc acctcgctaa cggattcacc actccaagaa ttggagccaa
     1861 tcaattcttg cggagaactg tgaatgcgca aaccaaccct tggcagaaca tatccatcgc
     1921 gtccgccatc tccagcagcc gcacgcggcg catctcgggc agcgttgggt cctggccacg
     1981 ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac
     2041 tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac
     2101 gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga
     2161 aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg
     2221 gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat
     2281 ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa
     2341 ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat
     2401 cattaccccc atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa
     2461 aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact
     2521 caacgagctg gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga
     2581 tgagctttac cgcaggatcc gcgaaattgt aagcgttaat attttgttaa aattcgcgtt
     2641 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     2701 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     2761 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     2821 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     2881 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     2941 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3001 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3061 gcggatcctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     3121 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     3181 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     3241 gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     3301 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc
     3361 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3421 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3481 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3541 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3601 cccgacagga ctataaagat accagggttt ccccctggaa gctccctcgt gcgctctcct
     3661 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3721 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3781 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3841 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3901 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3961 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4021 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4081 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4141 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4201 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4261 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4321 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4381 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4441 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4501 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4561 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg
     4621 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4681 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4741 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4801 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4861 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat
     4921 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4981 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5041 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5101 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5161 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5221 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5281 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     5341 aggccctttc gtcttcaaga attcccgaca ccatcgaatg gtgcaaaacc tttcgcggta
     5401 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
     5461 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     5521 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     5581 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     5641 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     5701 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     5761 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     5821 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     5881 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
     5941 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     6001 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
     6061 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
     6121 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     6181 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     6241 cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
     6301 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     6361 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     6421 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6481 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     6541 aatgtaagtt agctcactca ttaggcacaa ttctcatgtt tgacagctta tcatcgactg
     6601 cacggtgcac caatgcttct ggcgtcaggc agccatcgga agctgtggta tggctgtgca
     6661 ggtcgtaaat cactgcataa ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt
     6721 ttttgcgccg acatcataac ggttctggca aatattctga aatgagctgt tgacaattaa
     6781 tcatcggctc gtataatgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     6841 cagtccgttt aggtgttttc acgagcactt caccaacaag gaccatagat tatgaaaatc
     6901 gaagaa
//
