LOCUS       sequence-191138-        6972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            MPP1-1/1-IS2/2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6972)
  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 6972)
  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_MPP1-1/1-IS2/2
FEATURES             Location/Qualifiers
     source          1..6972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     13..31
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        54..131
                     /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             132..1214
                     /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(151..170)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1457..1462,1463..1478,1479..1485)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1493..1509
                     /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     1498..1520
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1594..1611
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1627..2280
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2238..2260
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2287..2310
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2323..2340
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2623..2670
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4263..4718)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4400..4421)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4612..4631
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4760..4782)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4868..5008
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4923..4942
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5023..5040)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5194..5781)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5952..6743,6744..6812))
                     /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     6575..6594
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6813..6917)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt cccgacacca
       61 tcgaatggtg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
      121 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
      181 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
      241 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
      301 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
      361 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
      421 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
      481 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
      541 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
      601 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
      661 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
      721 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
      781 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
      841 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
      901 ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg
      961 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
     1021 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
     1081 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
     1141 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     1201 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcacaattc
     1261 tcatgtttga cagcttatca tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc
     1321 catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca
     1381 aggcgcactc ccgttctgga taatgttttt tgcgccgaca tcataacggt tctggcaaat
     1441 attctgaaat gagctgttga caattaatca tcggctcgta taatgtgtgg aattgtgagc
     1501 ggataacaat ttcacacagg aaacagccag tccgtttagg tgttttcacg agcacttcac
     1561 caacaaggac catagattat gaaaatcgaa gaacaccatc accatcacca ttccagcggt
     1621 aagcttatgt cccctatact aggttattgg aaaattaagg gccttgtgca acccactcga
     1681 cttcttttgg aatatcttga agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt
     1741 gataaatggc gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct tccttattat
     1801 attgatggtg atgttaaatt aacacagtct atggccatca tacgttatat agctgacaag
     1861 cacaacatgt tgggtggttg tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg
     1921 gttttggata ttagatacgg tgtttcgaga attgcatata gtaaagactt tgaaactctc
     1981 aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt
     2041 cataaaacat atttaaatgg tgatcatgta acccatcctg acttcatgtt gtatgacgct
     2101 cttgatgttg ttttatacat ggacccaatg tgcctggatg cgttcccaaa attagtttgt
     2161 tttaaaaaac gtattgaagc tatcccacaa attgataagt acttgaaatc cagcaagtat
     2221 atagcatggc ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa
     2281 tcggatctgg aagttctgtt ccaggggccc ctgtccagcg gtctggttcc gcgtggttcc
     2341 ggtaccgcgg cccagccggc caaagcaatg aaagaaacga aaggcatgat ctccctgaaa
     2401 gttatcccga atcaacagtc ccgtctgccg gctctgcaag aagctggcct gaaatttgct
     2461 accggcgata tcatccagat catcaacaaa gatgactcta attggtggca aggccgtgtg
     2521 gaaggtagct ctaaagaatc agccggtctg attccgtcgc cggaactgca ggaatggcgc
     2581 gttgcgagta tggcccaatc cgcaccggcg gccgcataat aaaccgatac aattaaaggc
     2641 tccttttgga gccttttttt tggagatttt caacgtgaaa aaattattat tcgcaattcc
     2701 tttagttgtt cctttctatt ctcactccgc tgaaactgtt gaaagttgtt tagcaaaacc
     2761 ccatacagaa aattcattta ctaacgtctg gaaagacgac aaaactttag atcgttacgc
     2821 taactatgag ggctgtctgt ggaatgctac aggcgttgta gtttgtactg gtgacgaaac
     2881 tcagtgtcta gctagagtgg cggtggctct ggttccggtg attttgatta tgaaaagatg
     2941 gcaaacgcta ataagggggc tatgaccgaa aatgccgatg aaaacgcgct acagtctgac
     3001 gctaaaggca aacttgattc tgtcgctact gattacggtg ctgctatcga tggtttcatt
     3061 ggtgacgttt ccggccttgc taatggtaat ggtgctactg gtgattttgc tggctctaat
     3121 tcccaaatgg ctcaagtcgg tgacggtgat aattcacctt taatgaataa tttccgtcaa
     3181 tatttacctt ccctccctca atcggttgaa tgtcgccctt ttgtctttgg cgctggtaaa
     3241 ccatatgaat tttctattga ttgtgacaaa ataaacttat tccgtggtgt ctttgcgttt
     3301 cttttatatg ttgccacctt tatgtatgta ttttctacgt ttgctaacat actgcgtaat
     3361 aaggagtctt aatcatgcca gttcttttgg ctagcgccgc cctatacctt gtctgcctcc
     3421 ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac ctgaatggaa gccggcggca
     3481 cctcgctaac ggattcacca ctccaagaat tggagccaat caattcttgc ggagaactgt
     3541 gaatgcgcaa accaaccctt ggcagaacat atccatcgcg tccgccatct ccagcagccg
     3601 cacgcggcgc atctcgggca gcgttgggtc ctggccacgg gtgcgcatga tcgtgctcct
     3661 gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga atgaatcacc
     3721 gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac
     3781 atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt cagcgccctg
     3841 caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg gaacacctac
     3901 atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt cccgccgcat
     3961 ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt catcatcagt
     4021 aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca tgaacagaaa
     4081 tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta acatggcccg
     4141 ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg acgcggatga
     4201 acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc gcaggatccg
     4261 cgaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     4321 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     4381 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     4441 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     4501 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     4561 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     4621 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     4681 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg cggatcctgc ctcgcgcgtt
     4741 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     4801 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     4861 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta
     4921 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     4981 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct
     5041 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5101 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5161 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5221 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5281 ccagggtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5341 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5401 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5461 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5521 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5581 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     5641 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5701 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5761 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5821 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5881 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5941 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6001 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6061 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6121 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6181 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6241 agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt
     6301 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6361 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6421 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6481 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6541 cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact
     6601 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6661 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6721 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6781 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6841 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6901 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     6961 attatcatga ca
//
