LOCUS       sequence-390543-        4564 bp    DNA     circular SYN 15-APR-2025
DEFINITION  E. coli expression of HtrA2(delta133)-6His.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4564)
  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 4564)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pET-20b_HtrA2(delta(133))_S306A_6His
FEATURES             Location/Qualifiers
     source          1..4564
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     224..243
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     477..494
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    509..648
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(574..593)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     734..756
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(750..941)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1450..1469
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1450..1468
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1500..1522
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2514..2531
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2584..2602)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2598..2645
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2682..3137
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2769..2788)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2979..3000
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3163..3267
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3268..3336,3337..4128)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3486..3505)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4299..4564,1..323)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
       61 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
      121 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
      181 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
      241 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
      301 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
      361 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
      421 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
      481 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
      541 gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
      601 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
      661 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
      721 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
      781 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc
      841 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
      901 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
      961 ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
     1021 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
     1081 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
     1141 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
     1201 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
     1261 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
     1321 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
     1381 gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga gatctcgatc
     1441 ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct agaaataatt
     1501 ttgtttaact ttaagaagga gatatacata tggccgtccc tagcccgccg cccgcttctc
     1561 cccggagtca gtacaacttc atcgcagatg tggtggagaa gacagcacct gccgtggtct
     1621 atatcgagat cctggaccgg caccctttct tgggccgcga ggtccctatc tcgaacggct
     1681 cagggttcgt ggtggctgcc gatgggctca ttgtcaccaa cgcccatgtg gtggctgatc
     1741 ggcgcagagt ccgtgtgaga ctgctaagcg gcgacacgta tgaggccgtg gtcacagctg
     1801 tggatcccgt ggcagacatc gcaacgctga ggattcagac taaggagcct ctccccacgc
     1861 tgcctctggg acgctcagct gatgtccggc aaggggagtt tgttgttgca atgggaagtc
     1921 cctttgcact gcagaacacg atcacatccg gcattgttag ctctgctcag cgtccagcca
     1981 gagacctggg acttccgcaa accaatgtgg aatacattca aactgatgca gctattgatt
     2041 ttggaaacgc gggaggtccc ctggttaacc tggatgggga ggtgattgga gtgaacacca
     2101 tgaaggtcac agctggaatc tcctttgcca tcccttctga tcgtcttcga gagtttctgc
     2161 atcgtgggga aaagaagaat tcctcctccg gaatcagtgg gtcccagcgg cgctacattg
     2221 gggtgatgat gctgaccctg agtcccagca tccttgctga actacagctt cgagaaccaa
     2281 gctttcccga tgttcagcat ggtgtactca tccataaagt catcctgggc tcccctgcac
     2341 accgggctgg tctgcggcct ggtgatgtga ttttggccat tggggagcag atggtacaaa
     2401 atgctgaaga tgtttatgaa gctgttcgaa cccaatccca gttggcagtg cagatccggc
     2461 ggggacgaga aacactgacc ttatatgtga cccctgaggt cacagaactc gagcaccacc
     2521 accaccacca ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
     2581 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     2641 ttttgctgaa aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc
     2701 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     2761 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2821 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     2881 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     2941 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3001 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3061 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3121 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3181 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3241 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3301 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3361 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3421 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3481 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     3541 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3601 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3661 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3721 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3781 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     3841 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     3901 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3961 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4021 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4081 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4141 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4201 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4261 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4321 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4381 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4441 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4501 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4561 ctta
//
