LOCUS       sequence-192610-        6894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            CTTN-1/1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6894)
  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 6894)
  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_CTTN-1/1
FEATURES             Location/Qualifiers
     source          1..6894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     10..28
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        51..128
                     /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             129..1211
                     /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(148..167)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1454..1459,1460..1475,1476..1482)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1490..1506
                     /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     1495..1517
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1591..1608
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1624..2277
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2235..2257
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2284..2307
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2320..2337
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2542..2589
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4182..4637)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4319..4340)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4531..4550
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4679..4701)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4787..4927
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4842..4861
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4942..4959)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5113..5700)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5871..6662,6663..6731))
                     /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     6494..6513
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6732..6836)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acctataaaa ataggcgtat cacgaggccc tttcgtcttc aagaattccc gacaccatcg
       61 aatggtgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg
      121 tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc
      181 agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag
      241 tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg
      301 gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc
      361 aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga
      421 tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac
      481 gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag
      541 ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca
      601 gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg
      661 gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc
      721 tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag
      781 gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg
      841 ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta
      901 ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag
      961 acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc
     1021 aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc
     1081 tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct
     1141 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     1201 gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc acaattctca
     1261 tgtttgacag cttatcatcg actgcacggt gcaccaatgc ttctggcgtc aggcagccat
     1321 cggaagctgt ggtatggctg tgcaggtcgt aaatcactgc ataattcgtg tcgctcaagg
     1381 cgcactcccg ttctggataa tgttttttgc gccgacatca taacggttct ggcaaatatt
     1441 ctgaaatgag ctgttgacaa ttaatcatcg gctcgtataa tgtgtggaat tgtgagcgga
     1501 taacaatttc acacaggaaa cagccagtcc gtttaggtgt tttcacgagc acttcaccaa
     1561 caaggaccat agattatgaa aatcgaagaa caccatcacc atcaccattc cagcggtaag
     1621 cttatgtccc ctatactagg ttattggaaa attaagggcc ttgtgcaacc cactcgactt
     1681 cttttggaat atcttgaaga aaaatatgaa gagcatttgt atgagcgcga tgaaggtgat
     1741 aaatggcgaa acaaaaagtt tgaattgggt ttggagtttc ccaatcttcc ttattatatt
     1801 gatggtgatg ttaaattaac acagtctatg gccatcatac gttatatagc tgacaagcac
     1861 aacatgttgg gtggttgtcc aaaagagcgt gcagagattt caatgcttga aggagcggtt
     1921 ttggatatta gatacggtgt ttcgagaatt gcatatagta aagactttga aactctcaaa
     1981 gttgattttc ttagcaagct acctgaaatg ctgaaaatgt tcgaagatcg tttatgtcat
     2041 aaaacatatt taaatggtga tcatgtaacc catcctgact tcatgttgta tgacgctctt
     2101 gatgttgttt tatacatgga cccaatgtgc ctggatgcgt tcccaaaatt agtttgtttt
     2161 aaaaaacgta ttgaagctat cccacaaatt gataagtact tgaaatccag caagtatata
     2221 gcatggcctt tgcagggctg gcaagccacg tttggtggtg gcgaccatcc tccaaaatcg
     2281 gatctggaag ttctgttcca ggggcccctg tccagcggtc tggttccgcg tggttccggt
     2341 accgcggccc agccggccta caccgcagtt gccctgtatg attatcaagc cgccggtgat
     2401 gatgaaatct cctttgatcc ggatgacatt atcaccaaca ttgaaatgat tgatgatggt
     2461 tggtggcgtg gtgtttgtaa aggtcgttac ggtctctttc cagccaacta tgttgaactg
     2521 cgtcaggcgg ccgcataata aaccgataca attaaaggct ccttttggag cctttttttt
     2581 ggagattttc aacgtgaaaa aattattatt cgcaattcct ttagttgttc ctttctattc
     2641 tcactccgct gaaactgttg aaagttgttt agcaaaaccc catacagaaa attcatttac
     2701 taacgtctgg aaagacgaca aaactttaga tcgttacgct aactatgagg gctgtctgtg
     2761 gaatgctaca ggcgttgtag tttgtactgg tgacgaaact cagtgtctag ctagagtggc
     2821 ggtggctctg gttccggtga ttttgattat gaaaagatgg caaacgctaa taagggggct
     2881 atgaccgaaa atgccgatga aaacgcgcta cagtctgacg ctaaaggcaa acttgattct
     2941 gtcgctactg attacggtgc tgctatcgat ggtttcattg gtgacgtttc cggccttgct
     3001 aatggtaatg gtgctactgg tgattttgct ggctctaatt cccaaatggc tcaagtcggt
     3061 gacggtgata attcaccttt aatgaataat ttccgtcaat atttaccttc cctccctcaa
     3121 tcggttgaat gtcgcccttt tgtctttggc gctggtaaac catatgaatt ttctattgat
     3181 tgtgacaaaa taaacttatt ccgtggtgtc tttgcgtttc ttttatatgt tgccaccttt
     3241 atgtatgtat tttctacgtt tgctaacata ctgcgtaata aggagtctta atcatgccag
     3301 ttcttttggc tagcgccgcc ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat
     3361 ggagccgggc cacctcgacc tgaatggaag ccggcggcac ctcgctaacg gattcaccac
     3421 tccaagaatt ggagccaatc aattcttgcg gagaactgtg aatgcgcaaa ccaacccttg
     3481 gcagaacata tccatcgcgt ccgccatctc cagcagccgc acgcggcgca tctcgggcag
     3541 cgttgggtcc tggccacggg tgcgcatgat cgtgctcctg tcgttgagga cccggctagg
     3601 ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag
     3661 cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct tcggtttccg
     3721 tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt tccggatctg
     3781 catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgctg
     3841 gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca gttgtttacc
     3901 ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg tgagcatcct
     3961 ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca cggaggcatc
     4021 agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa gccagacatt
     4081 aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca tctgtgaatc
     4141 gcttcacgac cacgctgatg agctttaccg caggatccgc gaaattgtaa gcgttaatat
     4201 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4261 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4321 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4381 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4441 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4501 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4561 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4621 gccgctacag ggcgcgtcgc ggatcctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
     4681 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     4741 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
     4801 agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt
     4861 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     4921 catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4981 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5041 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5101 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5161 aagtcagagg tggcgaaacc cgacaggact ataaagatac cagggtttcc ccctggaagc
     5221 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5281 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5341 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5401 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5461 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5521 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     5581 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5641 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5701 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5761 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5821 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5881 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5941 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6001 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6061 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6121 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6181 attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6241 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6301 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6361 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6421 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6481 gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6541 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6601 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6661 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6721 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6781 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6841 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac atta
//
