LOCUS       sequence-396473-        6096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For protein purification of Csx29.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6096)
  AUTHORS   Strecker J, Demircioglu FE, Li D, Faure G, Wilkinson ME, Gootenberg 
            JS, Abudayyeh OO, Nishimasu H, Macrae RK, Zhang F
  TITLE     RNA-activated protein cleavage with a CRISPR-associated 
            endopeptidase.
  JOURNAL   Science. 2022 Nov 25;378(6622):874-881. doi: 
            10.1126/science.add7450. Epub 2022 Nov 3.
  PUBMED    36423276
REFERENCE   2  (bases 1 to 6096)
  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.add7450"; journalName: "Science"; date: 
            "2022-11-25- 25"; volume: "378"; issue: "6622"; pages: "874-881"
COMMENT     Sequence Label: pCDF_6xHIS_Csx29
FEATURES             Location/Qualifiers
     source          1..6096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      629..638
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        2464..2482
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2483..2507
                     /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)."
     CDS             2616..2660
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2698..2716)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2712..2759
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2930..3721)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(3722..3813)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(4874..5956)
                     /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     5918..5937
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5957..6034)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(6080..6096,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc ctatcagaga
      121 tatccaggac agactgaaga ccgccaagtt cgacaacaag gacgacatga tgaacctggc
      181 ttcaagcctg tacaagtacg agaaacagct gatggacagc tctgaggcta cactgtgcca
      241 gcagggcctg agcaaccggc ctaactcttt ttctcagctg agccaatttc gggacagcga
      301 cattcagagc aaggccggcg gccagaccgg taagttctgg cagaacgagt acgaggcttg
      361 taaaaacttc cagacacaca aggagagaag agaaaccctg gaacagatca tcagattcct
      421 gcaaaatggc gctgaggaaa aggacgctga tgatctgctg ctgaagaccc tggcccgagc
      481 ctacttccac cgtggactgc tgtatagacc taagggcttc agcgtgcctg cgaggaaggt
      541 ggaagccatg aaaaaggcca tcgcctactg cgaaatcatc cttgataaga atgaggaaga
      601 atcggaggct ctgagaatct ggctgtacgc cgccatggaa ctccggagat gcggcgagga
      661 gtatcctgag aactttgcgg agaagctgtt ctaccttgcc aacgacggct tcatcagcga
      721 gctgtacgac atccggctgt tcctggaata caccgagaga gaagaggaca acaacttcct
      781 ggacatgatc ctgcaggaga atcaggacag agaaagactg ttcgagctgt gtctgtacaa
      841 ggccagagcc tgttttcacc tgaaccagct gaacgacgtg cggatctacg gcgagagcgc
      901 aatcgacaac gcccctggcg ccttcgccga tcctttctgg gatgaactgg tggagttcat
      961 cagaatgctg cggaacaaga agtctgaact gtggaaggaa attgccatca aggcctggga
     1021 caagtgcaga gaaaaggaaa tgaaggtggg caacaacatc tacctttctt ggtactgggc
     1081 cagacagaga gagctgtacg acctggcctt catggcccaa gacggcatcg agaagaaaac
     1141 aagaatcgcc gatagcctga agagccggac caccctgaga atccaggagc tgaatgagct
     1201 gagaaaggat gcccaccgga agcagaaccg tcggctggaa gataagctgg atcggatcat
     1261 cgagcaggag aacgaggcca gagacggcgc ctacctgcgg agaaaccccc cctgcttcac
     1321 cggcggaaaa agagaggaaa tcccctttgc cagacttcca cagaactgga tcgccgtgca
     1381 cttctatctg aacgagctgg aaagccacga aggcggcaag ggcggccacg ccctgatcta
     1441 cgaccctcag aaggctgaga aagatcagtg gcaagacaag agcttcgact acaaagaact
     1501 gcacagaaaa ttcctcgagt ggcaggagaa ctacatcctg aacgaggagg gcagcgccga
     1561 tttcctggtg acactgtgca gagagatcga gaaagccatg cctttcctgt ttaagtccga
     1621 ggtgatcccc gaggacagac ccgtgctgtg gattcctcac ggcttcctgc atagactgcc
     1681 tctgcacgcc gctatgaaga gcggaaacaa ttctaatatc gagatcttct gggagcggca
     1741 cgccagcaga tatctccctg cttggcacct gttcgatcct gccccataca gccgcgagga
     1801 aagctccacc ctgctgaaga acttcgagga gtacgacttc cagaacctgg aaaacggcga
     1861 aatcgaggtc tacgccccca gcagccccaa gaaggtgaag gaagccatcc gggaaaaccc
     1921 cgctattctg ctgctgctgt gtcacggcga ggccgacatg acaaaccctt tccggagctg
     1981 cctgaagctg aaaaacaaag atatgaccat cttcgacctg ctgaccgtgg aagatgtgag
     2041 gctgtctggc agcagaatcc ttctgggagc ctgcgagtct gacatggtgc cacctctgga
     2101 gttcagcgtg gacgaacacc tgagcgtgtc cggcgctttt ctcagccata aggccggaga
     2161 aatcgtcgcc ggcctgtgga ccgtggactc tgagaaggtt gacgagtgct acagctacct
     2221 ggtggaagag aaggacttcc tgagaaacct gcaggagtgg cagatggccg agacagagaa
     2281 cttcagaagc gagaatgata gctccctgtt ctacaagatc gccccatttc ggatcattgg
     2341 ctttcctgcc gaatgagatc cgaattcgag ctcggcgcgc ctgcaggccg acaagcttgc
     2401 ggccgcataa tgcttaagtc gaacagaaag taatcgtatt gtacacggcc gcataatcga
     2461 aattaatacg actcactata ggggaattgt gagcggataa caattcccca tcttagtata
     2521 ttagttaagt ataagaagga gatatacata tggcagatct caattggata tcggccggcc
     2581 acgcgatcgc tgacgtcggt accctcgagt ctggtaaaga aaccgctgct gcgaaatttg
     2641 aacgccagca catggactcg tctactagcg cagcttaatt aacctaggct gctgccaccg
     2701 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     2761 tgaaacctca ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg
     2821 taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg acgaccgggt
     2881 catcgtggcc ggatcttgcg gcccctcggc ttgaacgaat tgttagacat tatttgccga
     2941 ctaccttggt gatctcgcct ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg
     3001 aggccaagcg atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct
     3061 gatactgggc cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt
     3121 tgccggttac tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc
     3181 cagcccagtc gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat
     3241 cctgttcagg aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat
     3301 gttctcttgc ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga
     3361 tacctgcaag aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat
     3421 aacgccacgg aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct
     3481 cgctctctcc aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg
     3541 tttcatcaag ccttacggtc accgtaacca gcaaatcaat atcactgtgt ggcttcaggc
     3601 cgccatccac tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct
     3661 cgatgacgcc aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccctca
     3721 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     3781 acatatttga atgtatttag aaaaataaac aaatagctag ctcactcggt cgctacgctc
     3841 cgggcgtgag actgcggcgg gcgctgcgga cacatacaaa gttacccaca gattccgtgg
     3901 ataagcaggg gactaacatg tgaggcaaaa cagcagggcc gcgccggtgg cgtttttcca
     3961 taggctccgc cctcctgcca gagttcacat aaacagacgc ttttccggtg catctgtggg
     4021 agccgtgagg ctcaaccatg aatctgacag tacgggcgaa acccgacagg acttaaagat
     4081 ccccaccgtt tccggcgggt cgctccctct tgcgctctcc tgttccgacc ctgccgttta
     4141 ccggatacct gttccgcctt tctcccttac gggaagtgtg gcgctttctc atagctcaca
     4201 cactggtatc tcggctcggt gtaggtcgtt cgctccaagc tgggctgtaa gcaagaactc
     4261 cccgttcagc ccgactgctg cgccttatcc ggtaactgtt cacttgagtc caacccggaa
     4321 aagcacggta aaacgccact ggcagcagcc attggtaact gggagttcgc agaggatttg
     4381 tttagctaaa cacgcggttg ctcttgaagt gtgcgccaaa gtccggctac actggaagga
     4441 cagatttggt tgctgtgctc tgcgaaagcc agttaccacg gttaagcagt tccccaactg
     4501 acttaacctt cgatcaaacc acctccccag gtggtttttt cgaaggttaa gtcagttggg
     4561 gaactgctta accttcgatc aaaccacctc cccaggtggt tttttcgttt acagggcaaa
     4621 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct actgaaccgc
     4681 tctagatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat
     4741 ataagttgta attctcatgt tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     4801 gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt
     4861 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4921 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     4981 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     5041 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     5101 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg
     5161 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     5221 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     5281 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     5341 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     5401 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     5461 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     5521 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     5581 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     5641 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     5701 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     5761 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     5821 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     5881 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     5941 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     6001 taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc
     6061 gactcctgca ttaggaaatt aatacgactc actata
//
