LOCUS       sequence-191134-        6954 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            DOCK5-1/1-IS1/2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6954)
  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 6954)
  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_DOCK5-1/1-IS1/2
FEATURES             Location/Qualifiers
     source          1..6954
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..23
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        46..123
                     /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             124..1206
                     /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(143..162)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1449..1454,1455..1470,1471..1477)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1485..1501
                     /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     1490..1512
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1586..1603
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1619..2272
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2230..2252
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2279..2302
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2315..2332
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2597..2644
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4237..4692)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4374..4395)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4586..4605
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4734..4756)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4842..4982
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4897..4916
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4997..5014)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5168..5755)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5926..6717,6718..6786))
                     /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     6549..6568
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6787..6891)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taaaaatagg cgtatcacga ggccctttcg tcttcaagaa ttcccgacac catcgaatgg
       61 tgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
      121 aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc
      181 gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa
      241 gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa
      301 cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt
      361 gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta
      421 gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc
      481 agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc
      541 tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt
      601 attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac
      661 cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct
      721 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
      781 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
      841 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
      901 tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc
      961 tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
     1021 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
     1081 cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc
     1141 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     1201 cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcacaat tctcatgttt
     1261 gacagcttat catcgactgc acggtgcacc aatgcttctg gcgtcaggca gccatcggaa
     1321 gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct caaggcgcac
     1381 tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa atattctgaa
     1441 atgagctgtt gacaattaat catcggctcg tataatgtgt ggaattgtga gcggataaca
     1501 atttcacaca ggaaacagcc agtccgttta ggtgttttca cgagcacttc accaacaagg
     1561 accatagatt atgaaaatcg aagaacacca tcaccatcac cattccagcg gtaagcttat
     1621 gtcccctata ctaggttatt ggaaaattaa gggccttgtg caacccactc gacttctttt
     1681 ggaatatctt gaagaaaaat atgaagagca tttgtatgag cgcgatgaag gtgataaatg
     1741 gcgaaacaaa aagtttgaat tgggtttgga gtttcccaat cttccttatt atattgatgg
     1801 tgatgttaaa ttaacacagt ctatggccat catacgttat atagctgaca agcacaacat
     1861 gttgggtggt tgtccaaaag agcgtgcaga gatttcaatg cttgaaggag cggttttgga
     1921 tattagatac ggtgtttcga gaattgcata tagtaaagac tttgaaactc tcaaagttga
     1981 ttttcttagc aagctacctg aaatgctgaa aatgttcgaa gatcgtttat gtcataaaac
     2041 atatttaaat ggtgatcatg taacccatcc tgacttcatg ttgtatgacg ctcttgatgt
     2101 tgttttatac atggacccaa tgtgcctgga tgcgttccca aaattagttt gttttaaaaa
     2161 acgtattgaa gctatcccac aaattgataa gtacttgaaa tccagcaagt atatagcatg
     2221 gcctttgcag ggctggcaag ccacgtttgg tggtggcgac catcctccaa aatcggatct
     2281 ggaagttctg ttccaggggc ccctgtccag cggtctggtt ccgcgtggtt ccggtaccgc
     2341 ggcccagccg gcctggattc cgacgaaacg tcaaaaatat ggcgtggcga tctataacta
     2401 caatgcgtca caagatgttg aactgagcct gcaaattggc gataccgtgc atattctgga
     2461 aatgtatgaa ggctggtatc gtggttacac gctgcagaac aaaagcaaaa aaggcatttt
     2521 tccggaaacc tacatccatc tgaaagaagc gacggttgaa gacctgggtc aacacgaaac
     2581 cgcggccgca taataaaccg atacaattaa aggctccttt tggagccttt tttttggaga
     2641 ttttcaacgt gaaaaaatta ttattcgcaa ttcctttagt tgttcctttc tattctcact
     2701 ccgctgaaac tgttgaaagt tgtttagcaa aaccccatac agaaaattca tttactaacg
     2761 tctggaaaga cgacaaaact ttagatcgtt acgctaacta tgagggctgt ctgtggaatg
     2821 ctacaggcgt tgtagtttgt actggtgacg aaactcagtg tctagctaga gtggcggtgg
     2881 ctctggttcc ggtgattttg attatgaaaa gatggcaaac gctaataagg gggctatgac
     2941 cgaaaatgcc gatgaaaacg cgctacagtc tgacgctaaa ggcaaacttg attctgtcgc
     3001 tactgattac ggtgctgcta tcgatggttt cattggtgac gtttccggcc ttgctaatgg
     3061 taatggtgct actggtgatt ttgctggctc taattcccaa atggctcaag tcggtgacgg
     3121 tgataattca cctttaatga ataatttccg tcaatattta ccttccctcc ctcaatcggt
     3181 tgaatgtcgc ccttttgtct ttggcgctgg taaaccatat gaattttcta ttgattgtga
     3241 caaaataaac ttattccgtg gtgtctttgc gtttctttta tatgttgcca cctttatgta
     3301 tgtattttct acgtttgcta acatactgcg taataaggag tcttaatcat gccagttctt
     3361 ttggctagcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc
     3421 cgggccacct cgacctgaat ggaagccggc ggcacctcgc taacggattc accactccaa
     3481 gaattggagc caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac ccttggcaga
     3541 acatatccat cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg ggcagcgttg
     3601 ggtcctggcc acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc
     3661 ggggttgcct tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact
     3721 gctgctgcaa aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt
     3781 cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg atctgcatcg
     3841 caggatgctg ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctggcatt
     3901 gaccctgagt gatttttctc tggtcccgcc gcatccatac cgccagttgt ttaccctcac
     3961 aacgttccag taaccgggca tgttcatcat cagtaacccg tatcgtgagc atcctctctc
     4021 gtttcatcgg tatcattacc cccatgaaca gaaatccccc ttacacggag gcatcagtga
     4081 ccaaacagga aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc
     4141 ttctggagaa actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc
     4201 acgaccacgc tgatgagctt taccgcagga tccgcgaaat tgtaagcgtt aatattttgt
     4261 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4321 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4381 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4441 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4501 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4561 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4621 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4681 tacagggcgc gtcgcggatc ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     4741 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     4801 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
     4861 cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag attgtactga
     4921 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     4981 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5041 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5101 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5161 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5221 agaggtggcg aaacccgaca ggactataaa gataccaggg tttccccctg gaagctccct
     5281 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5341 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5401 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5461 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5521 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5581 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     5641 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5701 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5761 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5821 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5881 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5941 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6001 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6061 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6121 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6181 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6241 tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6301 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6361 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6421 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6481 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6541 aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6601 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6661 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6721 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6781 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6841 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6901 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa ccta
//
