LOCUS       sequence-191116-        6975 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            MYO15A-2/2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6975)
  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 6975)
  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_MYO15A-2/2
FEATURES             Location/Qualifiers
     source          1..6975
                     /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      1033..1080
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(2673..3128)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2810..2831)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3022..3041
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3170..3192)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3278..3418
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3333..3352
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3433..3450)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3604..4191)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4362..5153,5154..5222))
                     /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     4985..5004
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5223..5327)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5395..5413
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5436..5513
                     /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             5514..6596
                     /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(5533..5552)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6839..6844,6845..6860,6861..6867)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    6875..6891
                     /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     6880..6902
                     /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 agccggccga ctacgtcgtc
      781 gctgttcgca attttctgcc ggaagacccg gctctgctgg cgtttcataa aggtgacatc
      841 attcatctgc aaccgctgga accgccgcgt gtgggttata gcgcaggttg cgtggttcgt
      901 cgcaaagtcg tgtacctgga agaactgcgt cgccgtggcc cggattttgg ttggcgtttc
      961 ggcaccattc atggccgcgt tggtcgtttt ccgtctgaac tggtccagcc ggcggccgcg
     1021 gccgcataat aaaccgatac aattaaaggc tccttttgga gccttttttt tggagatttt
     1081 caacgtgaaa aaattattat tcgcaattcc tttagttgtt cctttctatt ctcactccgc
     1141 tgaaactgtt gaaagttgtt tagcaaaacc ccatacagaa aattcattta ctaacgtctg
     1201 gaaagacgac aaaactttag atcgttacgc taactatgag ggctgtctgt ggaatgctac
     1261 aggcgttgta gtttgtactg gtgacgaaac tcagtgtcta gctagagtgg cggtggctct
     1321 ggttccggtg attttgatta tgaaaagatg gcaaacgcta ataagggggc tatgaccgaa
     1381 aatgccgatg aaaacgcgct acagtctgac gctaaaggca aacttgattc tgtcgctact
     1441 gattacggtg ctgctatcga tggtttcatt ggtgacgttt ccggccttgc taatggtaat
     1501 ggtgctactg gtgattttgc tggctctaat tcccaaatgg ctcaagtcgg tgacggtgat
     1561 aattcacctt taatgaataa tttccgtcaa tatttacctt ccctccctca atcggttgaa
     1621 tgtcgccctt ttgtctttgg cgctggtaaa ccatatgaat tttctattga ttgtgacaaa
     1681 ataaacttat tccgtggtgt ctttgcgttt cttttatatg ttgccacctt tatgtatgta
     1741 ttttctacgt ttgctaacat actgcgtaat aaggagtctt aatcatgcca gttcttttgg
     1801 ctagcgccgc cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg
     1861 ccacctcgac ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat
     1921 tggagccaat caattcttgc ggagaactgt gaatgcgcaa accaaccctt ggcagaacat
     1981 atccatcgcg tccgccatct ccagcagccg cacgcggcgc atctcgggca gcgttgggtc
     2041 ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg
     2101 ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg
     2161 ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta
     2221 aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg
     2281 atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc
     2341 ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg
     2401 ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt
     2461 catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa
     2521 acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat taacgcttct
     2581 ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga
     2641 ccacgctgat gagctttacc gcaggatccg cgaaattgta agcgttaata ttttgttaaa
     2701 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     2761 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     2821 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     2881 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     2941 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3001 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3061 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3121 gggcgcgtcg cggatcctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     3181 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     3241 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
     3301 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt
     3361 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     3421 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3481 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3541 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3601 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3661 gtggcgaaac ccgacaggac tataaagata ccagggtttc cccctggaag ctccctcgtg
     3721 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3781 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3841 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3901 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3961 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4021 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     4081 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4141 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4201 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4261 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4321 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4381 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4441 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4501 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4561 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4621 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca
     4681 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4741 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4801 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4861 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4921 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca
     4981 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5041 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5101 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5161 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5221 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5281 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5341 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     5401 cgtatcacga ggccctttcg tcttcaagaa ttcccgacac catcgaatgg tgcaaaacct
     5461 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
     5521 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
     5581 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
     5641 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
     5701 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
     5761 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
     5821 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
     5881 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
     5941 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
     6001 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
     6061 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
     6121 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca
     6181 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc
     6241 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc
     6301 gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata
     6361 tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc
     6421 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac
     6481 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg
     6541 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     6601 aacgcaatta atgtaagtta gctcactcat taggcacaat tctcatgttt gacagcttat
     6661 catcgactgc acggtgcacc aatgcttctg gcgtcaggca gccatcggaa gctgtggtat
     6721 ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct caaggcgcac tcccgttctg
     6781 gataatgttt tttgcgccga catcataacg gttctggcaa atattctgaa atgagctgtt
     6841 gacaattaat catcggctcg tataatgtgt ggaattgtga gcggataaca atttcacaca
     6901 ggaaacagcc agtccgttta ggtgttttca cgagcacttc accaacaagg accatagatt
     6961 atgaaaatcg aagaa
//
