LOCUS       sequence-192404-        6990 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            ZDHHC6-1/1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6990)
  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 6990)
  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_ZDHHC6-1/1
FEATURES             Location/Qualifiers
     source          1..6990
                     /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      2643..2690
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4283..4738)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4420..4441)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4632..4651
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4780..4802)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4888..5028
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4943..4962
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5043..5060)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5214..5801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5972..6763,6764..6832))
                     /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     6595..6614
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6833..6937)
                     /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 ggtccgttac aaagtcattg
     2401 aagattactc cggtgcatgt tgtccactga acaaaggtat caaaaccttc tttacctccc
     2461 cgtgtactga agaaccacgt attcagctgc aaaaaggtga atttatcctg gcaacccgtg
     2521 gtctgcgtta ttggctgtat ggtgacaaaa tcctggatga ttccttcatt gaaggtgtta
     2581 gccgtattcg tggttggttt ccgcgtaaat gcgttgaaaa atgcccggcg gccgcataat
     2641 aaaccgatac aattaaaggc tccttttgga gccttttttt tggagatttt caacgtgaaa
     2701 aaattattat tcgcaattcc tttagttgtt cctttctatt ctcactccgc tgaaactgtt
     2761 gaaagttgtt tagcaaaacc ccatacagaa aattcattta ctaacgtctg gaaagacgac
     2821 aaaactttag atcgttacgc taactatgag ggctgtctgt ggaatgctac aggcgttgta
     2881 gtttgtactg gtgacgaaac tcagtgtcta gctagagtgg cggtggctct ggttccggtg
     2941 attttgatta tgaaaagatg gcaaacgcta ataagggggc tatgaccgaa aatgccgatg
     3001 aaaacgcgct acagtctgac gctaaaggca aacttgattc tgtcgctact gattacggtg
     3061 ctgctatcga tggtttcatt ggtgacgttt ccggccttgc taatggtaat ggtgctactg
     3121 gtgattttgc tggctctaat tcccaaatgg ctcaagtcgg tgacggtgat aattcacctt
     3181 taatgaataa tttccgtcaa tatttacctt ccctccctca atcggttgaa tgtcgccctt
     3241 ttgtctttgg cgctggtaaa ccatatgaat tttctattga ttgtgacaaa ataaacttat
     3301 tccgtggtgt ctttgcgttt cttttatatg ttgccacctt tatgtatgta ttttctacgt
     3361 ttgctaacat actgcgtaat aaggagtctt aatcatgcca gttcttttgg ctagcgccgc
     3421 cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac
     3481 ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat tggagccaat
     3541 caattcttgc ggagaactgt gaatgcgcaa accaaccctt ggcagaacat atccatcgcg
     3601 tccgccatct ccagcagccg cacgcggcgc atctcgggca gcgttgggtc ctggccacgg
     3661 gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact
     3721 ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg
     3781 tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa
     3841 acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg
     3901 ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt
     3961 tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac
     4021 cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc
     4081 attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa
     4141 accgccctta acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc
     4201 aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat
     4261 gagctttacc gcaggatccg cgaaattgta agcgttaata ttttgttaaa attcgcgtta
     4321 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     4381 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     4441 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     4501 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     4561 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     4621 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     4681 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg
     4741 cggatcctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     4801 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     4861 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
     4921 agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     4981 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
     5041 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5101 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5161 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5221 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5281 ccgacaggac tataaagata ccagggtttc cccctggaag ctccctcgtg cgctctcctg
     5341 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5401 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5461 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5521 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5581 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5641 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5701 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5761 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5821 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5881 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5941 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6001 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6061 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6121 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6181 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6241 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg
     6301 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6361 ttacatgatc ccccatattg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6421 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6481 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6541 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata
     6601 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6661 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6721 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6781 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6841 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6901 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6961 ctgacgtcta agaaaccatt attatcatga
//
