LOCUS       sequence-191125-        6894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            SH3D21-1/3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6894)
  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 6894)
  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_SH3D21-1/3
FEATURES             Location/Qualifiers
     source          1..6894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..33
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        56..133
                     /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             134..1216
                     /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(153..172)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1459..1464,1465..1480,1481..1487)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1495..1511
                     /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     1500..1522
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1596..1613
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1629..2282
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2240..2262
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2289..2312
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2325..2342
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2547..2594
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4187..4642)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4324..4345)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4536..4555
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4684..4706)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4792..4932
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4847..4866
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4947..4964)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5118..5705)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5876..6667,6668..6736))
                     /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     6499..6518
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6737..6841)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa ttcccgacac
       61 catcgaatgg tgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt
      121 cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc
      181 ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga
      241 aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact
      301 ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc
      361 gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt
      421 gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc
      481 gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt
      541 ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat
      601 caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc
      661 attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct
      721 gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg
      781 ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg
      841 catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc
      901 cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac
      961 cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct
     1021 ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa
     1081 tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac
     1141 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     1201 ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcacaat
     1261 tctcatgttt gacagcttat catcgactgc acggtgcacc aatgcttctg gcgtcaggca
     1321 gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct
     1381 caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa
     1441 atattctgaa atgagctgtt gacaattaat catcggctcg tataatgtgt ggaattgtga
     1501 gcggataaca atttcacaca ggaaacagcc agtccgttta ggtgttttca cgagcacttc
     1561 accaacaagg accatagatt atgaaaatcg aagaacacca tcaccatcac cattccagcg
     1621 gtaagcttat gtcccctata ctaggttatt ggaaaattaa gggccttgtg caacccactc
     1681 gacttctttt ggaatatctt gaagaaaaat atgaagagca tttgtatgag cgcgatgaag
     1741 gtgataaatg gcgaaacaaa aagtttgaat tgggtttgga gtttcccaat cttccttatt
     1801 atattgatgg tgatgttaaa ttaacacagt ctatggccat catacgttat atagctgaca
     1861 agcacaacat gttgggtggt tgtccaaaag agcgtgcaga gatttcaatg cttgaaggag
     1921 cggttttgga tattagatac ggtgtttcga gaattgcata tagtaaagac tttgaaactc
     1981 tcaaagttga ttttcttagc aagctacctg aaatgctgaa aatgttcgaa gatcgtttat
     2041 gtcataaaac atatttaaat ggtgatcatg taacccatcc tgacttcatg ttgtatgacg
     2101 ctcttgatgt tgttttatac atggacccaa tgtgcctgga tgcgttccca aaattagttt
     2161 gttttaaaaa acgtattgaa gctatcccac aaattgataa gtacttgaaa tccagcaagt
     2221 atatagcatg gcctttgcag ggctggcaag ccacgtttgg tggtggcgac catcctccaa
     2281 aatcggatct ggaagttctg ttccaggggc ccctgtccag cggtctggtt ccgcgtggtt
     2341 ccggtaccgc ggcccagccg gccgaagtcc tggtcctggc gggttatcgt gctcagaaag
     2401 aagatgaact gtccctggct ccgggtgatg tggtccgtca agtccgttgg gtgccggcac
     2461 gtggttggct gcgtggtgaa tttggcggtc gttatggcct gttcccggaa cgcctggttc
     2521 aggaaattcc ggcggccgca taataaaccg atacaattaa aggctccttt tggagccttt
     2581 tttttggaga ttttcaacgt gaaaaaatta ttattcgcaa ttcctttagt tgttcctttc
     2641 tattctcact ccgctgaaac tgttgaaagt tgtttagcaa aaccccatac agaaaattca
     2701 tttactaacg tctggaaaga cgacaaaact ttagatcgtt acgctaacta tgagggctgt
     2761 ctgtggaatg ctacaggcgt tgtagtttgt actggtgacg aaactcagtg tctagctaga
     2821 gtggcggtgg ctctggttcc ggtgattttg attatgaaaa gatggcaaac gctaataagg
     2881 gggctatgac cgaaaatgcc gatgaaaacg cgctacagtc tgacgctaaa ggcaaacttg
     2941 attctgtcgc tactgattac ggtgctgcta tcgatggttt cattggtgac gtttccggcc
     3001 ttgctaatgg taatggtgct actggtgatt ttgctggctc taattcccaa atggctcaag
     3061 tcggtgacgg tgataattca cctttaatga ataatttccg tcaatattta ccttccctcc
     3121 ctcaatcggt tgaatgtcgc ccttttgtct ttggcgctgg taaaccatat gaattttcta
     3181 ttgattgtga caaaataaac ttattccgtg gtgtctttgc gtttctttta tatgttgcca
     3241 cctttatgta tgtattttct acgtttgcta acatactgcg taataaggag tcttaatcat
     3301 gccagttctt ttggctagcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg
     3361 tgcatggagc cgggccacct cgacctgaat ggaagccggc ggcacctcgc taacggattc
     3421 accactccaa gaattggagc caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac
     3481 ccttggcaga acatatccat cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg
     3541 ggcagcgttg ggtcctggcc acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg
     3601 ctaggctggc ggggttgcct tactggttag cagaatgaat caccgatacg cgagcgaacg
     3661 tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa caacatgaat ggtcttcggt
     3721 ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg
     3781 atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt attaacgaag
     3841 cgctggcatt gaccctgagt gatttttctc tggtcccgcc gcatccatac cgccagttgt
     3901 ttaccctcac aacgttccag taaccgggca tgttcatcat cagtaacccg tatcgtgagc
     3961 atcctctctc gtttcatcgg tatcattacc cccatgaaca gaaatccccc ttacacggag
     4021 gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat cagaagccag
     4081 acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc agacatctgt
     4141 gaatcgcttc acgaccacgc tgatgagctt taccgcagga tccgcgaaat tgtaagcgtt
     4201 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4261 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4321 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     4381 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     4441 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     4501 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     4561 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4621 aatgcgccgc tacagggcgc gtcgcggatc ctgcctcgcg cgtttcggtg atgacggtga
     4681 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     4741 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     4801 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag
     4861 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     4921 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4981 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5041 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5101 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5161 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggg tttccccctg
     5221 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5281 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5341 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5401 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5461 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5521 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     5581 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5641 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5701 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5761 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5821 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5881 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5941 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6001 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6061 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6121 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6181 ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6241 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6301 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6361 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6421 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6481 gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6541 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6601 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6661 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6721 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6781 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6841 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atga
//
