LOCUS       sequence-192401-        6921 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            TP53BP2-1/1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6921)
  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 6921)
  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_TP53BP2-1/1
FEATURES             Location/Qualifiers
     source          1..6921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     32..50
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        73..150
                     /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             151..1233
                     /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(170..189)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1476..1481,1482..1497,1498..1504)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1512..1528
                     /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     1517..1539
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1613..1630
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1646..2299
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2257..2279
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2306..2329
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2342..2359
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2591..2638
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4231..4686)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4368..4389)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4580..4599
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4728..4750)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4836..4976
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4891..4910
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4991..5008)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5162..5749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5920..6711,6712..6780))
                     /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     6543..6562
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6781..6885)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
       61 tcaagaattc ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc
      121 ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca
      181 gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt
      241 tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac
      301 cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt
      361 ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg
      421 ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg
      481 gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac
      541 caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc
      601 tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc
      661 gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt
      721 tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt
      781 cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg
      841 caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg
      901 ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta
      961 gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa
     1021 caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc
     1081 caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg
     1141 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     1201 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct
     1261 cactcattag gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat
     1321 gcttctggcg tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact
     1381 gcataattcg tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat
     1441 cataacggtt ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat
     1501 aatgtgtgga attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt
     1561 gttttcacga gcacttcacc aacaaggacc atagattatg aaaatcgaag aacaccatca
     1621 ccatcaccat tccagcggta agcttatgtc ccctatacta ggttattgga aaattaaggg
     1681 ccttgtgcaa cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt
     1741 gtatgagcgc gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt
     1801 tcccaatctt ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat
     1861 acgttatata gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat
     1921 ttcaatgctt gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag
     1981 taaagacttt gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat
     2041 gttcgaagat cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga
     2101 cttcatgttg tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc
     2161 gttcccaaaa ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta
     2221 cttgaaatcc agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg
     2281 tggcgaccat cctccaaaat cggatctgga agttctgttc caggggcccc tgtccagcgg
     2341 tctggttccg cgtggttccg gtaccgcggc ccagccggcc atggcccagg gcgcgctggg
     2401 tgttatttac gccctgtggg attatgaacc gcagaacgat gatgaactgc cgatgaaaga
     2461 aggtgattgt atgaccatta tccatcgtga agatgaagat gaaattgaat ggtggtgggc
     2521 acgtctgaat gataaagaag gttacgtccc acgtaatctg cttggtcttt atccagcggc
     2581 cgcataataa accgatacaa ttaaaggctc cttttggagc cttttttttg gagattttca
     2641 acgtgaaaaa attattattc gcaattcctt tagttgttcc tttctattct cactccgctg
     2701 aaactgttga aagttgttta gcaaaacccc atacagaaaa ttcatttact aacgtctgga
     2761 aagacgacaa aactttagat cgttacgcta actatgaggg ctgtctgtgg aatgctacag
     2821 gcgttgtagt ttgtactggt gacgaaactc agtgtctagc tagagtggcg gtggctctgg
     2881 ttccggtgat tttgattatg aaaagatggc aaacgctaat aagggggcta tgaccgaaaa
     2941 tgccgatgaa aacgcgctac agtctgacgc taaaggcaaa cttgattctg tcgctactga
     3001 ttacggtgct gctatcgatg gtttcattgg tgacgtttcc ggccttgcta atggtaatgg
     3061 tgctactggt gattttgctg gctctaattc ccaaatggct caagtcggtg acggtgataa
     3121 ttcaccttta atgaataatt tccgtcaata tttaccttcc ctccctcaat cggttgaatg
     3181 tcgccctttt gtctttggcg ctggtaaacc atatgaattt tctattgatt gtgacaaaat
     3241 aaacttattc cgtggtgtct ttgcgtttct tttatatgtt gccaccttta tgtatgtatt
     3301 ttctacgttt gctaacatac tgcgtaataa ggagtcttaa tcatgccagt tcttttggct
     3361 agcgccgccc tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc
     3421 acctcgacct gaatggaagc cggcggcacc tcgctaacgg attcaccact ccaagaattg
     3481 gagccaatca attcttgcgg agaactgtga atgcgcaaac caacccttgg cagaacatat
     3541 ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat ctcgggcagc gttgggtcct
     3601 ggccacgggt gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt
     3661 gccttactgg ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct
     3721 gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa
     3781 gtctggaaac gcggaagtca gcgccctgca ccattatgtt ccggatctgc atcgcaggat
     3841 gctgctggct accctgtgga acacctacat ctgtattaac gaagcgctgg cattgaccct
     3901 gagtgatttt tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt
     3961 ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca
     4021 tcggtatcat tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac
     4081 aggaaaaaac cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg
     4141 agaaactcaa cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc
     4201 acgctgatga gctttaccgc aggatccgcg aaattgtaag cgttaatatt ttgttaaaat
     4261 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     4321 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     4381 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     4441 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     4501 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     4561 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     4621 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     4681 gcgcgtcgcg gatcctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     4741 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     4801 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
     4861 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     4921 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
     4981 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5041 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5101 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5161 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5221 ggcgaaaccc gacaggacta taaagatacc agggtttccc cctggaagct ccctcgtgcg
     5281 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5341 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5401 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5461 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5521 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5581 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     5641 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5701 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5761 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5821 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5881 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5941 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6001 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6061 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6121 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6181 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg
     6241 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6301 aaggcgagtt acatgatccc ccatattgtg caaaaaagcg gttagctcct tcggtcctcc
     6361 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6421 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6481 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg
     6541 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6601 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6661 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6721 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6781 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6841 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6901 agtgccacct gacgtctaag a
//
