LOCUS       sequence-191788-        6978 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            NOXA1-1/2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6978)
  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 6978)
  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_NOXA1-1/2
FEATURES             Location/Qualifiers
     source          1..6978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     34..52
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        75..152
                     /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             153..1235
                     /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(172..191)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1478..1483,1484..1499,1500..1506)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1514..1530
                     /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     1519..1541
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1615..1632
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1648..2301
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2259..2281
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2308..2331
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2344..2361
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2650..2697
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4290..4745)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4427..4448)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4639..4658
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4787..4809)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4895..5035
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4950..4969
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5050..5067)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5221..5808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5979..6770,6771..6839))
                     /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     6602..6621
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6840..6944)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
       61 cttcaagaat tcccgacacc atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc
      121 gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg
      181 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg
      241 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca
      301 accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca
      361 gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac
      421 tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg
      481 cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg
      541 accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg
      601 tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg
      661 gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa
      721 gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa
      781 ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca
      841 tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg
      901 cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg
      961 tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca
     1021 aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg
     1081 gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc
     1141 tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     1201 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag
     1261 ctcactcatt aggcacaatt ctcatgtttg acagcttatc atcgactgca cggtgcacca
     1321 atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca
     1381 ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg ataatgtttt ttgcgccgac
     1441 atcataacgg ttctggcaaa tattctgaaa tgagctgttg acaattaatc atcggctcgt
     1501 ataatgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcca gtccgtttag
     1561 gtgttttcac gagcacttca ccaacaagga ccatagatta tgaaaatcga agaacaccat
     1621 caccatcacc attccagcgg taagcttatg tcccctatac taggttattg gaaaattaag
     1681 ggccttgtgc aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat
     1741 ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag
     1801 tttcccaatc ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc
     1861 atacgttata tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag
     1921 atttcaatgc ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat
     1981 agtaaagact ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa
     2041 atgttcgaag atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct
     2101 gacttcatgt tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat
     2161 gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag
     2221 tacttgaaat ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt
     2281 ggtggcgacc atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgtccagc
     2341 ggtctggttc cgcgtggttc cggtaccgcg gcccagccgg ccggtcgccc ggtcctgtac
     2401 caagtcgtgg cacaacattc atactctgca caaggtccgg aagatctggg ttttcgtcaa
     2461 ggtgatacgg tcgatgtgct gtgcgaagaa ccggatgttc cgctggcagt cgaccaggca
     2521 tggctggaag gtcattgcga cggtcgtatt ggtatctttc cgaaatgttt cgtggttccg
     2581 gcaggtccgc gtatgagcgg tgcaccgggt cgtctgccgc gctctcagca aggtgcggcc
     2641 gcataataaa ccgatacaat taaaggctcc ttttggagcc ttttttttgg agattttcaa
     2701 cgtgaaaaaa ttattattcg caattccttt agttgttcct ttctattctc actccgctga
     2761 aactgttgaa agttgtttag caaaacccca tacagaaaat tcatttacta acgtctggaa
     2821 agacgacaaa actttagatc gttacgctaa ctatgagggc tgtctgtgga atgctacagg
     2881 cgttgtagtt tgtactggtg acgaaactca gtgtctagct agagtggcgg tggctctggt
     2941 tccggtgatt ttgattatga aaagatggca aacgctaata agggggctat gaccgaaaat
     3001 gccgatgaaa acgcgctaca gtctgacgct aaaggcaaac ttgattctgt cgctactgat
     3061 tacggtgctg ctatcgatgg tttcattggt gacgtttccg gccttgctaa tggtaatggt
     3121 gctactggtg attttgctgg ctctaattcc caaatggctc aagtcggtga cggtgataat
     3181 tcacctttaa tgaataattt ccgtcaatat ttaccttccc tccctcaatc ggttgaatgt
     3241 cgcccttttg tctttggcgc tggtaaacca tatgaatttt ctattgattg tgacaaaata
     3301 aacttattcc gtggtgtctt tgcgtttctt ttatatgttg ccacctttat gtatgtattt
     3361 tctacgtttg ctaacatact gcgtaataag gagtcttaat catgccagtt cttttggcta
     3421 gcgccgccct ataccttgtc tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca
     3481 cctcgacctg aatggaagcc ggcggcacct cgctaacgga ttcaccactc caagaattgg
     3541 agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc aacccttggc agaacatatc
     3601 catcgcgtcc gccatctcca gcagccgcac gcggcgcatc tcgggcagcg ttgggtcctg
     3661 gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc cggctaggct ggcggggttg
     3721 ccttactggt tagcagaatg aatcaccgat acgcgagcga acgtgaagcg actgctgctg
     3781 caaaacgtct gcgacctgag caacaacatg aatggtcttc ggtttccgtg tttcgtaaag
     3841 tctggaaacg cggaagtcag cgccctgcac cattatgttc cggatctgca tcgcaggatg
     3901 ctgctggcta ccctgtggaa cacctacatc tgtattaacg aagcgctggc attgaccctg
     3961 agtgattttt ctctggtccc gccgcatcca taccgccagt tgtttaccct cacaacgttc
     4021 cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg agcatcctct ctcgtttcat
     4081 cggtatcatt acccccatga acagaaatcc cccttacacg gaggcatcag tgaccaaaca
     4141 ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc cagacattaa cgcttctgga
     4201 gaaactcaac gagctggacg cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca
     4261 cgctgatgag ctttaccgca ggatccgcga aattgtaagc gttaatattt tgttaaaatt
     4321 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     4381 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     4441 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     4501 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     4561 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     4621 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     4681 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4741 cgcgtcgcgg atcctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     4801 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     4861 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
     4921 atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     4981 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc
     5041 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5101 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5161 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5221 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5281 gcgaaacccg acaggactat aaagatacca gggtttcccc ctggaagctc cctcgtgcgc
     5341 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5401 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5461 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5521 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5581 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5641 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5701 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5761 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5821 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5881 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5941 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6001 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6061 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6121 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6181 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6241 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc
     6301 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6361 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6421 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6481 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6541 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg
     6601 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6661 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6721 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6781 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6841 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6901 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6961 gtgccacctg acgtctaa
//
