LOCUS       sequence-201853-        6924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            NOXA1-2/2-SV2/2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6924)
  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 6924)
  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_NOXA1-2/2-SV2/2
FEATURES             Location/Qualifiers
     source          1..6924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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     42..64
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             138..155
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             171..824
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     782..804
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             831..854
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             867..884
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      1119..1166
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(2759..3214)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2896..2917)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3108..3127
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3256..3278)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3364..3504
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3419..3438
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3519..3536)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3690..4277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4448..5239,5240..5308))
                     /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     5071..5090
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5309..5413)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5481..5499
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5522..5599
                     /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             5600..6682
                     /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(5619..5638)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag ccagtccgtt taggtgtttt cacgagcact tcaccaacaa ggaccataga
      121 ttatgaaaat cgaagaacac catcaccatc accattccag cggtaagctt atgtccccta
      181 tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt ttggaatatc
      241 ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa tggcgaaaca
      301 aaaagtttga attgggtttg gagtttccca atcttcctta ttatattgat ggtgatgtta
      361 aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac atgttgggtg
      421 gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg gatattagat
      481 acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt gattttctta
      541 gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa acatatttaa
      601 atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat gttgttttat
      661 acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa aaacgtattg
      721 aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca tggcctttgc
      781 agggctggca agccacgttt ggtggtggcg accatcctcc aaaatcggat ctggaagttc
      841 tgttccaggg gcccctgtcc agcggtctgg ttccgcgtgg ttccggtacc gcggcccagc
      901 cggccatggc ccagggcgcg ctgtatcaag ttgtcgccca gcattcttat tctgcccagg
      961 gtccagaaga tttgggtttc cgtcagggtg ataccgttga tgttctgtgc gaagaaccag
     1021 atgttccact ggcagttgat caagcatggc tggaaggtca ttgcgatggt cgtattggta
     1081 tctttccgaa atgttttgtt gttccggccg cataataaac cgatacaatt aaaggctcct
     1141 tttggagcct tttttttgga gattttcaac gtgaaaaaat tattattcgc aattccttta
     1201 gttgttcctt tctattctca ctccgctgaa actgttgaaa gttgtttagc aaaaccccat
     1261 acagaaaatt catttactaa cgtctggaaa gacgacaaaa ctttagatcg ttacgctaac
     1321 tatgagggct gtctgtggaa tgctacaggc gttgtagttt gtactggtga cgaaactcag
     1381 tgtctagcta gagtggcggt ggctctggtt ccggtgattt tgattatgaa aagatggcaa
     1441 acgctaataa gggggctatg accgaaaatg ccgatgaaaa cgcgctacag tctgacgcta
     1501 aaggcaaact tgattctgtc gctactgatt acggtgctgc tatcgatggt ttcattggtg
     1561 acgtttccgg ccttgctaat ggtaatggtg ctactggtga ttttgctggc tctaattccc
     1621 aaatggctca agtcggtgac ggtgataatt cacctttaat gaataatttc cgtcaatatt
     1681 taccttccct ccctcaatcg gttgaatgtc gcccttttgt ctttggcgct ggtaaaccat
     1741 atgaattttc tattgattgt gacaaaataa acttattccg tggtgtcttt gcgtttcttt
     1801 tatatgttgc cacctttatg tatgtatttt ctacgtttgc taacatactg cgtaataagg
     1861 agtcttaatc atgccagttc ttttggctag cgccgcccta taccttgtct gcctccccgc
     1921 gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga atggaagccg gcggcacctc
     1981 gctaacggat tcaccactcc aagaattgga gccaatcaat tcttgcggag aactgtgaat
     2041 gcgcaaacca acccttggca gaacatatcc atcgcgtccg ccatctccag cagccgcacg
     2101 cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt gctcctgtcg
     2161 ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata
     2221 cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga
     2281 atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc
     2341 attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct
     2401 gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat
     2461 accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc
     2521 cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc
     2581 ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt
     2641 atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag
     2701 gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag gatccgcgaa
     2761 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     2821 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     2881 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     2941 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     3001 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3061 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3121 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3181 cccgccgcgc ttaatgcgcc gctacagggc gcgtcgcgga tcctgcctcg cgcgtttcgg
     3241 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     3301 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     3361 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg
     3421 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     3481 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3541 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3601 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3661 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3721 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3781 ggtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3841 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3901 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3961 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4021 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4081 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     4141 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4201 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4261 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4321 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4381 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4441 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4501 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4561 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4621 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4681 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4741 tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4801 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atattgtgca
     4861 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4921 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4981 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5041 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
     5101 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5161 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5221 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5281 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5341 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5401 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     5461 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattcc
     5521 cgacaccatc gaatggtgca aaacctttcg cggtatggca tgatagcgcc cggaagagag
     5581 tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg
     5641 tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac
     5701 gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca
     5761 acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca
     5821 cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt
     5881 ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct
     5941 tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat
     6001 tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac
     6061 acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct
     6121 ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc
     6181 gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc
     6241 ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa
     6301 tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat
     6361 gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga
     6421 cgataccgaa gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg
     6481 cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa
     6541 gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac
     6601 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     6661 ccgactggaa agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg
     6721 cacaattctc atgtttgaca gcttatcatc gactgcacgg tgcaccaatg cttctggcgt
     6781 caggcagcca tcggaagctg tggtatggct gtgcaggtcg taaatcactg cataattcgt
     6841 gtcgctcaag gcgcactccc gttctggata atgttttttg cgccgacatc ataacggttc
     6901 tggcaaatat tctgaaatga gctg
//
