LOCUS       sequence-389057-        5858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli expression of SMAC(delta(55))-9His.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5858)
  AUTHORS   Hunkeler M, Jin CY, Fischer ES
  TITLE     Structures of BIRC6-client complexes provide a mechanism of 
            SMAC-mediated release of caspases.
  JOURNAL   Science. 2023 Mar 17;379(6637):1105-1111. doi: 
            10.1126/science.ade5750. Epub 2023 Feb 9.
  PUBMED    36758104
REFERENCE   2  (bases 1 to 5858)
  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: 
            "10.1126/science.ade5750"; journalName: "Science"; date: 
            "2023-03-17- 17"; volume: "379"; issue: "6637"; pages: "1105-1111"
COMMENT     Sequence Label: pNIC-Bio2_SMAC(delta(55))_9His
FEATURES             Location/Qualifiers
     source          1..5858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             644..670
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     CDS             723..740
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(793..811)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      807..854
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      891..1346
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(978..997)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1188..1209
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1438..2253)
                     /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(1438..2253)
                     /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     2161..2180
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2245..2264
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2375..2963
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2864..2883
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3117..3134
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3149..3288
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3214..3233)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3374..3396
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3390..3581)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4390..5472)
                     /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
                     EACKAAVHNLLAQRVSGLIINYQLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5434..5453
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5473..5550)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5756..5775
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggccgttcct atcgcgcaga aatctgaacc
      121 tcattccttg tcgtcggaag ctttgatgcg tcgtgctgta tctttagtca cggattccac
      181 ctcaactttt ttgagccaaa caacatatgc gcttattgaa gcgattacag aatacactaa
      241 agcggtgtat actctgacta gtttgtatcg ccagtatact tcccttcttg gaaaaatgaa
      301 tagtgaagaa gaggatgaag tatggcaagt tattatcgga gcccgcgcgg aaatgactag
      361 taaacatcag gagtacttga agttagaaac cacctggatg accgctgtag ggttgagtga
      421 gatggctgct gaagcggcct atcagactgg agccgatcag gcatcaatta cagcacgtaa
      481 ccacatccaa ttagtgaagc tgcaagtcga agaagtacac caactttcgc gcaaagcaga
      541 gaccaaattg gccgaggctc aaattgagga gctgcgccaa aaaacccagg aagaggggga
      601 agaacgtgca gaatcggagc aggaggctta tcttcgtgag gatcaccatc atcatcatca
      661 tcaccatcat tgataaggat ccgaattcga gctccgtcga caagcttgcg gccgcactcg
      721 agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt
      781 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
      841 tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg
      901 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
      961 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     1021 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     1081 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     1141 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     1201 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     1261 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     1321 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     1381 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa ttaattctta
     1441 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
     1501 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag
     1561 gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat
     1621 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
     1681 atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc
     1741 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc
     1801 ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
     1861 caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc
     1921 ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc
     1981 aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag
     2041 tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
     2101 ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt
     2161 atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta atcgcggcct
     2221 agagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
     2281 agcagacagt tttattgttc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     2341 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2401 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2461 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2521 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2581 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     2641 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     2701 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     2761 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     2821 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     2881 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     2941 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3001 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     3061 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3121 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     3181 gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3241 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     3301 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     3361 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     3421 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc
     3481 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
     3541 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
     3601 ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
     3661 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
     3721 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
     3781 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
     3841 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
     3901 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
     3961 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
     4021 gacaggagca cgatcatgcg cacccgtggg gccgccatgc cggcgataat ggcctgcttc
     4081 tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt
     4141 ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg
     4201 aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc
     4261 ataagtgcgg cgacgatagt catgtcccgc gcccaccgga aggagctgac tgggttgaag
     4321 gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt
     4381 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4441 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
     4501 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
     4561 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
     4621 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc
     4681 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
     4741 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
     4801 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
     4861 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
     4921 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
     4981 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
     5041 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gctgatagtt
     5101 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
     5161 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
     5221 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
     5281 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
     5341 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     5401 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     5461 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
     5521 gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact
     5581 cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga
     5641 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     5701 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     5761 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     5821 gtccggcgta gaggatcgag atctcgatcc cgcgaaat
//
