LOCUS       sequence-192247-        6912 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            SH3D19-3/5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6912)
  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 6912)
  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_SH3D19-3/5
FEATURES             Location/Qualifiers
     source          1..6912
                     /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      2565..2612
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4205..4660)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4342..4363)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4554..4573
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4702..4724)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4810..4950
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4865..4884
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4965..4982)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5136..5723)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5894..6685,6686..6754))
                     /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     6517..6536
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6755..6859)
                     /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 gccatggccc agggcgcgct gtctcgttgt gttgcccgtt
     2401 tcgaatacat tggtgaacag aaagatgaac tgtcctttag cgaaggtgaa atcattatcc
     2461 tgaaagaata tgtcaacgaa gaatgggcac gtggtgaagt tcgtggtcgt accggtatct
     2521 ttccgctgaa ctttgttgaa ccggtcgaag cggccgcata ataaaccgat acaattaaag
     2581 gctccttttg gagccttttt tttggagatt ttcaacgtga aaaaattatt attcgcaatt
     2641 cctttagttg ttcctttcta ttctcactcc gctgaaactg ttgaaagttg tttagcaaaa
     2701 ccccatacag aaaattcatt tactaacgtc tggaaagacg acaaaacttt agatcgttac
     2761 gctaactatg agggctgtct gtggaatgct acaggcgttg tagtttgtac tggtgacgaa
     2821 actcagtgtc tagctagagt ggcggtggct ctggttccgg tgattttgat tatgaaaaga
     2881 tggcaaacgc taataagggg gctatgaccg aaaatgccga tgaaaacgcg ctacagtctg
     2941 acgctaaagg caaacttgat tctgtcgcta ctgattacgg tgctgctatc gatggtttca
     3001 ttggtgacgt ttccggcctt gctaatggta atggtgctac tggtgatttt gctggctcta
     3061 attcccaaat ggctcaagtc ggtgacggtg ataattcacc tttaatgaat aatttccgtc
     3121 aatatttacc ttccctccct caatcggttg aatgtcgccc ttttgtcttt ggcgctggta
     3181 aaccatatga attttctatt gattgtgaca aaataaactt attccgtggt gtctttgcgt
     3241 ttcttttata tgttgccacc tttatgtatg tattttctac gtttgctaac atactgcgta
     3301 ataaggagtc ttaatcatgc cagttctttt ggctagcgcc gccctatacc ttgtctgcct
     3361 ccccgcgttg cgtcgcggtg catggagccg ggccacctcg acctgaatgg aagccggcgg
     3421 cacctcgcta acggattcac cactccaaga attggagcca atcaattctt gcggagaact
     3481 gtgaatgcgc aaaccaaccc ttggcagaac atatccatcg cgtccgccat ctccagcagc
     3541 cgcacgcggc gcatctcggg cagcgttggg tcctggccac gggtgcgcat gatcgtgctc
     3601 ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca
     3661 ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca
     3721 acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc
     3781 tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct
     3841 acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc
     3901 atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca
     3961 gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga
     4021 aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc
     4081 cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat
     4141 gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcaggatc
     4201 cgcgaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     4261 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     4321 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     4381 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     4441 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     4501 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     4561 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     4621 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cgcggatcct gcctcgcgcg
     4681 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     4741 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     4801 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac
     4861 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     4921 agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg
     4981 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5041 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5101 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5161 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5221 taccagggtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5281 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5341 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5401 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5461 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5521 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     5581 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5641 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5701 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5761 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5821 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5881 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5941 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6001 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6061 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6121 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6181 atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg
     6241 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatat
     6301 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6361 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6421 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6481 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     6541 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6601 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6661 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6721 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6781 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6841 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     6901 ttattatcat ga
//
