LOCUS       sequence-386053-        6294 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  protein expression of the GST-tagged BIR3 domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6294)
  AUTHORS   Schwalm MP, Berger LM, Meuter MN, Vasta JD, Corona CR, Rohm S, 
            Berger BT, Farges F, Beinert SM, Preuss F, Morasch V, Rogov VV, 
            Mathea S, Saxena K, Robers MB, Muller S, Knapp S
  TITLE     A Toolbox for the Generation of Chemical Probes for Baculovirus IAP 
            Repeat Containing Proteins.
  JOURNAL   Front Cell Dev Biol. 2022 May 26;10:886537. doi: 
            10.3389/fcell.2022.886537. eCollection 2022.
  PUBMED    35721509
REFERENCE   2  (bases 1 to 6294)
  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"; journalName: "Front Cell 
            Dev Biol."; date: "2022-05-26"; volume: "10:886537. doi"; pages: " 
            10.3389/fcell.2022.886537. eCollection 2022"
COMMENT     Sequence Label: pGTVL2-BIRC2-BIR3
FEATURES             Location/Qualifiers
     source          1..6294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     102..124
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(118..309)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1118..2200)
                     /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     2162..2181
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2201..2278)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2484..2503
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2587..2606
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2587..2605
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2606..2630
                     /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)."
     RBS             2645..2667
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2678..2695
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2702..3355
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGADHPPK"
     CDS             3383..3403
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3745..3762
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3815..3833)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3829..3876
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3913..4368
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4000..4019)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4210..4231
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4460..5275)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4460..5275)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5183..5202
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5267..5286
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5397..5985
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5886..5905
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6139..6156
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(6171..6294,1..16)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6236..6255)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
       61 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
      121 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
      181 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
      241 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
      301 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
      361 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
      421 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
      481 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca
      541 tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact
      601 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca
      661 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca
      721 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc
      781 cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac
      841 gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt
      901 cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc
      961 tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc
     1021 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc
     1081 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     1141 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     1201 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct
     1261 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc
     1321 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt
     1381 cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca
     1441 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat
     1501 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg
     1561 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg
     1621 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca
     1681 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg
     1741 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa
     1801 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa
     1861 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca
     1921 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt
     1981 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt
     2041 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg
     2101 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac
     2161 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac
     2221 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg
     2281 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg
     2341 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     2401 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     2461 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     2521 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg
     2581 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa
     2641 taattttgtt taactttaag aaggagatat acatatgcac catcatcatc atcattcttc
     2701 tatgtcccct atactaggtt attggaaaat taagggcctt gtgcaaccca ctcgacttct
     2761 tttggaatat cttgaagaaa aatatgaaga gcatttgtat gagcgcgatg aaggtgataa
     2821 atggcgaaac aaaaagtttg aattgggttt ggagtttccc aatcttcctt attatattga
     2881 tggtgatgtt aaattaacac agtctatggc catcatacgt tatatagctg acaagcacaa
     2941 catgttgggt ggttgtccaa aagagcgtgc agagatttca atgcttgaag gagcggtttt
     3001 ggatattaga tacggtgttt cgagaattgc atatagtaaa gactttgaaa ctctcaaagt
     3061 tgattttctt agcaagctac ctgaaatgct gaaaatgttc gaagatcgtt tatgtcataa
     3121 aacatattta aatggtgatc atgtaaccca tcctgacttc atgttgtatg acgctcttga
     3181 tgttgtttta tacatggacc caatgtgcct ggatgcgttc ccaaaattag tttgttttaa
     3241 aaaacgtatt gaagctatcc cacaaattga taagtacttg aaatccagca agtatatagc
     3301 atggcctttg cagggctggc aagccacgtt tggtggtgcc gaccatcctc caaaatcgag
     3361 ctcaggtgta gatctgggta ccgagaacct gtacttccaa tccatgagca tttcaaatct
     3421 gagcatgcag acacatgcag ctcgaatgag aacatttatg tactggccat ctagtgttcc
     3481 agttcagcct gagcagcttg caagtgctgg cttttattat gtgggtcgca atgatgatgt
     3541 caaatgcttt tgttgtgatg gtggcttgag gtgttgggaa tctggagatg atccatgggt
     3601 agaacatgcc aagtggtttc caaggtgtga gttcttgata cgaatgaaag gccaagagtt
     3661 tgttgatgag attcaaggta gatattaaag gtggatacgg atccgaattc gagctccgtc
     3721 gacaagcttg cggccgcact cgagcaccac caccaccacc actgagatcc ggctgctaac
     3781 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     3841 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     3901 ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3961 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4021 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4081 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4141 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4201 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4261 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4321 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     4381 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4441 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
     4501 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
     4561 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     4621 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     4681 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
     4741 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     4801 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
     4861 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     4921 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     4981 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     5041 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     5101 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     5161 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     5221 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
     5281 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
     5341 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5401 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5461 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5521 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     5581 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5641 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5701 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5761 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5821 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5881 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5941 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6001 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6061 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6121 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     6181 tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa
     6241 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtga
//
