LOCUS       sequence-192594-        6946 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            PIK3R2-1/1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6946)
  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 6946)
  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_PIK3R2-1/1
FEATURES             Location/Qualifiers
     source          1..6946
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      67..114
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(1707..2162)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1844..1865)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2056..2075
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(2204..2226)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2312..2452
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2367..2386
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2467..2484)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2638..3225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(3396..4187,4188..4256))
                     /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     4019..4038
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4257..4361)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4429..4447
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4470..4547
                     /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             4548..5630
                     /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(4567..4586)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5873..5878,5879..5894,5895..5901)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    5909..5925
                     /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     5914..5936
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             6010..6027
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6043..6696
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     6654..6676
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             6703..6726
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6739..6756
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
ORIGIN
        1 gtacccgtca gcgtggtgat tttcccggtn acttacgtcg aatacattgg tgcggccgca
       61 taataaaccg atacaattaa aggctccttt tggagccttt tttttggaga ttttcaacgt
      121 gaaaaaatta ttattcgcaa ttcctttagt tgttcctttc tattctcact ccgctgaaac
      181 tgttgaaagt tgtttagcaa aaccccatac agaaaattca tttactaacg tctggaaaga
      241 cgacaaaact ttagatcgtt acgctaacta tgagggctgt ctgtggaatg ctacaggcgt
      301 tgtagtttgt actggtgacg aaactcagtg tctagctaga gtggcggtgg ctctggttcc
      361 ggtgattttg attatgaaaa gatggcaaac gctaataagg gggctatgac cgaaaatgcc
      421 gatgaaaacg cgctacagtc tgacgctaaa ggcaaacttg attctgtcgc tactgattac
      481 ggtgctgcta tcgatggttt cattggtgac gtttccggcc ttgctaatgg taatggtgct
      541 actggtgatt ttgctggctc taattcccaa atggctcaag tcggtgacgg tgataattca
      601 cctttaatga ataatttccg tcaatattta ccttccctcc ctcaatcggt tgaatgtcgc
      661 ccttttgtct ttggcgctgg taaaccatat gaattttcta ttgattgtga caaaataaac
      721 ttattccgtg gtgtctttgc gtttctttta tatgttgcca cctttatgta tgtattttct
      781 acgtttgcta acatactgcg taataaggag tcttaatcat gccagttctt ttggctagcg
      841 ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc cgggccacct
      901 cgacctgaat ggaagccggc ggcacctcgc taacggattc accactccaa gaattggagc
      961 caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac ccttggcaga acatatccat
     1021 cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg ggcagcgttg ggtcctggcc
     1081 acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct
     1141 tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa
     1201 aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct
     1261 ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg
     1321 ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt
     1381 gatttttctc tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag
     1441 taaccgggca tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg
     1501 tatcattacc cccatgaaca gaaatccccc ttacacggag gcatcagtga ccaaacagga
     1561 aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa
     1621 actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc
     1681 tgatgagctt taccgcagga tccgcgaaat tgtaagcgtt aatattttgt taaaattcgc
     1741 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     1801 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     1861 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     1921 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     1981 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     2041 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     2101 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     2161 gtcgcggatc ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     2221 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     2281 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
     2341 gcggagtgta tactggctta actatgcggc atcagagcag attgtactga gagtgcacca
     2401 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc
     2461 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2521 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2581 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2641 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2701 aaacccgaca ggactataaa gataccaggg tttccccctg gaagctccct cgtgcgctct
     2761 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2821 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2881 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2941 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3001 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3061 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3121 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3181 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3241 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3301 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3361 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3421 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3481 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3541 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3601 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3661 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc
     3721 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3781 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3841 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3901 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3961 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat
     4021 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4081 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4141 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4201 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4261 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4321 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4381 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     4441 acgaggccct ttcgtcttca agaattcccg acaccatcga atggtgcaaa acctttcgcg
     4501 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
     4561 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     4621 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
     4681 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
     4741 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
     4801 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
     4861 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
     4921 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg
     4981 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag
     5041 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
     5101 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc
     5161 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg
     5221 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg
     5281 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg
     5341 gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc
     5401 cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
     5461 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga
     5521 aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     5581 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     5641 attaatgtaa gttagctcac tcattaggca caattctcat gtttgacagc ttatcatcga
     5701 ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg gtatggctgt
     5761 gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat
     5821 gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc tgttgacaat
     5881 taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     5941 agccagtccg tttaggtgtt ttcacgagca cttcaccaac aaggaccata gattatgaaa
     6001 atcgaagaac accatcacca tcaccattcc agcggtaagc ttatgtcccc tatactaggt
     6061 tattggaaaa ttaagggcct tgtgcaaccc actcgacttc ttttggaata tcttgaagaa
     6121 aaatatgaag agcatttgta tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt
     6181 gaattgggtt tggagtttcc caatcttcct tattatattg atggtgatgt taaattaaca
     6241 cagtctatgg ccatcatacg ttatatagct gacaagcaca acatgttggg tggttgtcca
     6301 aaagagcgtg cagagatttc aatgcttgaa ggagcggttt tggatattag atacggtgtt
     6361 tcgagaattg catatagtaa agactttgaa actctcaaag ttgattttct tagcaagcta
     6421 cctgaaatgc tgaaaatgtt cgaagatcgt ttatgtcata aaacatattt aaatggtgat
     6481 catgtaaccc atcctgactt catgttgtat gacgctcttg atgttgtttt atacatggac
     6541 ccaatgtgcc tggatgcgtt cccaaaatta gtttgtttta aaaaacgtat tgaagctatc
     6601 ccacaaattg ataagtactt gaaatccagc aagtatatag catggccttt gcagggctgg
     6661 caagccacgt ttggtggtgg cgaccatcct ccaaaatcgg atctggaagt tctgttccag
     6721 gggcccctgt ccagcggtct ggttccgcgt ggttccggta ccgcggccca gccggcctat
     6781 cagtatcgtg ccctgtatcc atttcgtcgt gaacgtccgg aagatttaga actgctgccg
     6841 ggtgatgttt tagttgtttc tcgtgcagca ctgcaagcat taggtgttgc agaaggtggt
     6901 gaacgttgtc cacaatccgt tggttggatg ccaggtctga atgaac
//
