LOCUS       sequence-192273-        6936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            RIMBP2-3/3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6936)
  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 6936)
  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_RIMBP2-3/3
FEATURES             Location/Qualifiers
     source          1..6936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     28..46
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        69..146
                     /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             147..1229
                     /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(166..185)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1472..1477,1478..1493,1494..1500)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1508..1524
                     /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     1513..1535
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1609..1626
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1642..2295
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2253..2275
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2302..2325
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2338..2355
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2602..2649
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4242..4697)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4379..4400)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4591..4610
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4739..4761)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4847..4987
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4902..4921
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5002..5019)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5173..5760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5931..6722,6723..6791))
                     /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     6554..6573
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6792..6896)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa
       61 gaattcccga caccatcgaa tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg
      121 aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt
      181 atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg
      241 cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg
      301 tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg
      361 ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg
      421 ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc
      481 acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg
      541 atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg
      601 accagacacc catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg
      661 agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg
      721 tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc
      781 cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa
      841 tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg
      901 gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg
      961 gatacgacga taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg
     1021 attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg
     1081 cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc
     1141 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     1201 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact
     1261 cattaggcac aattctcatg tttgacagct tatcatcgac tgcacggtgc accaatgctt
     1321 ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat
     1381 aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata
     1441 acggttctgg caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg
     1501 tgtggaattg tgagcggata acaatttcac acaggaaaca gccagtccgt ttaggtgttt
     1561 tcacgagcac ttcaccaaca aggaccatag attatgaaaa tcgaagaaca ccatcaccat
     1621 caccattcca gcggtaagct tatgtcccct atactaggtt attggaaaat taagggcctt
     1681 gtgcaaccca ctcgacttct tttggaatat cttgaagaaa aatatgaaga gcatttgtat
     1741 gagcgcgatg aaggtgataa atggcgaaac aaaaagtttg aattgggttt ggagtttccc
     1801 aatcttcctt attatattga tggtgatgtt aaattaacac agtctatggc catcatacgt
     1861 tatatagctg acaagcacaa catgttgggt ggttgtccaa aagagcgtgc agagatttca
     1921 atgcttgaag gagcggtttt ggatattaga tacggtgttt cgagaattgc atatagtaaa
     1981 gactttgaaa ctctcaaagt tgattttctt agcaagctac ctgaaatgct gaaaatgttc
     2041 gaagatcgtt tatgtcataa aacatattta aatggtgatc atgtaaccca tcctgacttc
     2101 atgttgtatg acgctcttga tgttgtttta tacatggacc caatgtgcct ggatgcgttc
     2161 ccaaaattag tttgttttaa aaaacgtatt gaagctatcc cacaaattga taagtacttg
     2221 aaatccagca agtatatagc atggcctttg cagggctggc aagccacgtt tggtggtggc
     2281 gaccatcctc caaaatcgga tctggaagtt ctgttccagg ggcccctgtc cagcggtctg
     2341 gttccgcgtg gttccggtac cgcggcccag ccggccatgg cccagggcgc gctgcgtcgt
     2401 atggttgccc tgtatgatta tgatccgcgt gaatcctccc caaacgttga tgttgaagcc
     2461 gaactgacct tttgcaccgg tgacattatc accgtttttg gtgaaattga tgaagatggt
     2521 ttctactacg gtgaactgaa cggtcaaaaa ggtctggttc cgtccaactt tctggaagaa
     2581 gttccagcgg ccgcataata aaccgataca attaaaggct ccttttggag cctttttttt
     2641 ggagattttc aacgtgaaaa aattattatt cgcaattcct ttagttgttc ctttctattc
     2701 tcactccgct gaaactgttg aaagttgttt agcaaaaccc catacagaaa attcatttac
     2761 taacgtctgg aaagacgaca aaactttaga tcgttacgct aactatgagg gctgtctgtg
     2821 gaatgctaca ggcgttgtag tttgtactgg tgacgaaact cagtgtctag ctagagtggc
     2881 ggtggctctg gttccggtga ttttgattat gaaaagatgg caaacgctaa taagggggct
     2941 atgaccgaaa atgccgatga aaacgcgcta cagtctgacg ctaaaggcaa acttgattct
     3001 gtcgctactg attacggtgc tgctatcgat ggtttcattg gtgacgtttc cggccttgct
     3061 aatggtaatg gtgctactgg tgattttgct ggctctaatt cccaaatggc tcaagtcggt
     3121 gacggtgata attcaccttt aatgaataat ttccgtcaat atttaccttc cctccctcaa
     3181 tcggttgaat gtcgcccttt tgtctttggc gctggtaaac catatgaatt ttctattgat
     3241 tgtgacaaaa taaacttatt ccgtggtgtc tttgcgtttc ttttatatgt tgccaccttt
     3301 atgtatgtat tttctacgtt tgctaacata ctgcgtaata aggagtctta atcatgccag
     3361 ttcttttggc tagcgccgcc ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat
     3421 ggagccgggc cacctcgacc tgaatggaag ccggcggcac ctcgctaacg gattcaccac
     3481 tccaagaatt ggagccaatc aattcttgcg gagaactgtg aatgcgcaaa ccaacccttg
     3541 gcagaacata tccatcgcgt ccgccatctc cagcagccgc acgcggcgca tctcgggcag
     3601 cgttgggtcc tggccacggg tgcgcatgat cgtgctcctg tcgttgagga cccggctagg
     3661 ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag
     3721 cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct tcggtttccg
     3781 tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt tccggatctg
     3841 catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgctg
     3901 gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca gttgtttacc
     3961 ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg tgagcatcct
     4021 ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca cggaggcatc
     4081 agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa gccagacatt
     4141 aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca tctgtgaatc
     4201 gcttcacgac cacgctgatg agctttaccg caggatccgc gaaattgtaa gcgttaatat
     4261 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4321 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4381 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4441 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4501 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4561 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4621 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4681 gccgctacag ggcgcgtcgc ggatcctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
     4741 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     4801 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
     4861 agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt
     4921 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     4981 catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5041 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5101 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5161 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5221 aagtcagagg tggcgaaacc cgacaggact ataaagatac cagggtttcc ccctggaagc
     5281 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5341 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5401 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5461 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5521 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5581 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     5641 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5701 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5761 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5821 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5881 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5941 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6001 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6061 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6121 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6181 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6241 attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6301 tcccaacgat caaggcgagt tacatgatcc cccatattgt gcaaaaaagc ggttagctcc
     6361 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6421 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6481 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6541 gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6601 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6661 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6721 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6781 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6841 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6901 tttccccgaa aagtgccacc tgacgtctaa gaaacc
//
