LOCUS       sequence-191124-        6690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            C1orf168-1/1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6690)
  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 6690)
  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_C1orf168-1/1
FEATURES             Location/Qualifiers
     source          1..6690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(133..511)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     458..477
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(696..800)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     868..886
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        909..986
                     /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             987..2069
                     /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(1006..1025)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2312..2317,2318..2333,2334..2340)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2348..2364
                     /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     2353..2375
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             2449..2466
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2482..3135
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3093..3115
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             3142..3165
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3178..3195
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      3454..3501
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(5094..5549)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5231..5252)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5443..5462
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(5591..5613)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5699..5839
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5754..5773
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5854..5871)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6025..6612)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
       61 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
      121 aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt
      181 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
      241 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
      301 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
      361 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
      421 cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga
      481 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
      541 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
      601 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
      661 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
      721 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
      781 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
      841 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa
      901 gaattcccga caccatcgaa tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg
      961 aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt
     1021 atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg
     1081 cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg
     1141 tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg
     1201 ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg
     1261 ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc
     1321 acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg
     1381 atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg
     1441 accagacacc catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg
     1501 agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg
     1561 tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc
     1621 cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa
     1681 tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg
     1741 gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg
     1801 gatacgacga taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg
     1861 attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg
     1921 cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc
     1981 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2041 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact
     2101 cattaggcac aattctcatg tttgacagct tatcatcgac tgcacggtgc accaatgctt
     2161 ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat
     2221 aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata
     2281 acggttctgg caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg
     2341 tgtggaattg tgagcggata acaatttcac acaggaaaca gccagtccgt ttaggtgttt
     2401 tcacgagcac ttcaccaaca aggaccatag attatgaaaa tcgaagaaca ccatcaccat
     2461 caccattcca gcggtaagct tatgtcccct atactaggtt attggaaaat taagggcctt
     2521 gtgcaaccca ctcgacttct tttggaatat cttgaagaaa aatatgaaga gcatttgtat
     2581 gagcgcgatg aaggtgataa atggcgaaac aaaaagtttg aattgggttt ggagtttccc
     2641 aatcttcctt attatattga tggtgatgtt aaattaacac agtctatggc catcatacgt
     2701 tatatagctg acaagcacaa catgttgggt ggttgtccaa aagagcgtgc agagatttca
     2761 atgcttgaag gagcggtttt ggatattaga tacggtgttt cgagaattgc atatagtaaa
     2821 gactttgaaa ctctcaaagt tgattttctt agcaagctac ctgaaatgct gaaaatgttc
     2881 gaagatcgtt tatgtcataa aacatattta aatggtgatc atgtaaccca tcctgacttc
     2941 atgttgtatg acgctcttga tgttgtttta tacatggacc caatgtgcct ggatgcgttc
     3001 ccaaaattag tttgttttaa aaaacgtatt gaagctatcc cacaaattga taagtacttg
     3061 aaatccagca agtatatagc atggcctttg cagggctggc aagccacgtt tggtggtggc
     3121 gaccatcctc caaaatcgga tctggaagtt ctgttccagg ggcccctgtc cagcggtctg
     3181 gttccgcgtg gttccggtac cgcggcccag ccggccaaac gtgaagaaaa actgttccgt
     3241 gaacgcttca aatacgataa agaaatcatt gtcatcaaca ccgccgtcgc ctgctcgaat
     3301 aacagccgta acggcatttt tgatctgccg atctctccgg gtgaagaact ggaagtgatt
     3361 gacaccacgg aacagaacct ggttatctgc cgcaatagta aaggcaaata tggttacgtc
     3421 ctgattgaac atctggatgc ggccgcataa taaaccgata caattaaagg ctccttttgg
     3481 agcctttttt ttggagattt tcaacgtgaa aaaattatta ttcgcaattc ctttagttgt
     3541 tcctttctat tctcactccg ctgaaactgt tgaaagttgt ttagcaaaac cccatacaga
     3601 aaattcattt actaacgtct ggaaagacga caaaacttta gatcgttacg ctaactatga
     3661 gggctgtctg tggaatgcta caggcgttgt agtttgtact ggtgacgaaa ctcagtgtct
     3721 agctagagtg gcggtggctc tggttccggt gattttgatt atgaaaagat ggcaaacgct
     3781 aataaggggg ctatgaccga aaatgccgat gaaaacgcgc tacagtctga cgctaaaggc
     3841 aaacttgatt ctgtcgctac tgattacggt gctgctatcg atggtttcat tggtgacgtt
     3901 tccggccttg ctaatggtaa tggtgctact ggtgattttg ctggctctaa ttcccaaatg
     3961 gctcaagtcg gtgacggtga taattcacct ttaatgaata atttccgtca atatttacct
     4021 tccctccctc aatcggttga atgtcgccct tttgtctttg gcgctggtaa accatatgaa
     4081 ttttctattg attgtgacaa aataaactta ttccgtggtg tctttgcgtt tcttttatat
     4141 gttgccacct ttatgtatgt attttctacg tttgctaaca tactgcgtaa taaggagtct
     4201 taatcatgcc agttcttttg gctagcgccg ccctatacct tgtctgcctc cccgcgttgc
     4261 gtcgcggtgc atggagccgg gccacctcga cctgaatgga agccggcggc acctcgctaa
     4321 cggattcacc actccaagaa ttggagccaa tcaattcttg cggagaactg tgaatgcgca
     4381 aaccaaccct tggcagaaca tatccatcgc gtccgccatc tccagcagcc gcacgcggcg
     4441 catctcgggc agcgttgggt cctggccacg ggtgcgcatg atcgtgctcc tgtcgttgag
     4501 gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga
     4561 gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt
     4621 cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct gcaccattat
     4681 gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt
     4741 aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca tccataccgc
     4801 cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag taacccgtat
     4861 cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa atccccctta
     4921 cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag
     4981 aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga
     5041 catctgtgaa tcgcttcacg accacgctga tgagctttac cgcaggatcc gcgaaattgt
     5101 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     5161 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     5221 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     5281 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     5341 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     5401 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     5461 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     5521 cgcgcttaat gcgccgctac agggcgcgtc gcggatcctg cctcgcgcgt ttcggtgatg
     5581 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     5641 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg
     5701 cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc
     5761 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     5821 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5881 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5941 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6001 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6061 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accagggttt
     6121 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6181 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6241 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6301 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6361 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6421 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     6481 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6541 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6601 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6661 aactcacgtt aagggatttt ggtcatgaga
//
