LOCUS       sequence-410199-        4069 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein purification vector to produce AcrIC8 in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4069)
  AUTHORS   Hu C, Myers MT, Zhou X, Hou Z, Lozen ML, Nam KH, Zhang Y, Ke A
  TITLE     Exploiting activation and inactivation mechanisms in type I-C 
            CRISPR-Cas3 for genome-editing applications.
  JOURNAL   Mol Cell. 2024 Feb 1;84(3):463-475.e5. doi: 
            10.1016/j.molcel.2023.12.034. Epub 2024 Jan 18.
  PUBMED    38242128
REFERENCE   2  (bases 1 to 4069)
  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.molcel.2023.12.034"; journalName: "Mol Cell"; date: 
            "2024-02-1- 1"; volume: "84"; issue: "3"; pages: "463-475.e5"
COMMENT     Sequence Label: pRSFDuet-3C-AcrIC8
FEATURES             Location/Qualifiers
     source          1..4069
                     /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"
     CDS             107..130
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     promoter        453..471
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    472..496
                     /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             606..650
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(688..706)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      702..749
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(979..1794)
                     /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(979..1794)
                     /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     1702..1721
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1795..1886)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1902..2651)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(1912..1929)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2152..2171)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2847..3929)
                     /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     3891..3910
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3930..4007)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4053..4069,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccagctgg aagttctgtt
      121 ccaggggccc ctgggatcca tgtacgcaat ccgcaaaatc caattcttct acgggcctac
      181 tgacaaaaaa tcctatgttg gcgaggaggc tggtggccgc cgcgaacttt tcaaaacgcg
      241 agccgaggcg caggcacgta tcgaggacct ggaggagggc gtgtactacc tcgcgcacaa
      301 cgagtcgggc cgtcccgact acaaaatcgt ctgggtgcga ggcgaagccc agtttgagca
      361 cgcccgctgg atgcgcggct gaagcttgcg gccgcataat gcttaagtcg aacagaaagt
      421 aatcgtattg tacacggccg cataatcgaa attaatacga ctcactatag gggaattgtg
      481 agcggataac aattccccat cttagtatat tagttaagta taagaaggag atatacatat
      541 ggccagatct caattggata tcggccggcc acgcgatcgc tgacgtcggt accctcgagt
      601 ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg
      661 cagcttaatt aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg
      721 cctctaaacg ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg
      781 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt
      841 aacgaccctg ccctgaaccg acgacaagct gacgaccggg tctccgcaag tggcactttt
      901 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      961 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     1021 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     1081 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     1141 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     1201 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
     1261 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     1321 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cggtcgctgt taaaaggaca
     1381 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     1441 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     1501 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     1561 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     1621 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     1681 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     1741 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcatactctt
     1801 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     1861 tgaatgtatt tagaaaaata aacaaatagg catgcagcgc tcttccgctt cctcgctcac
     1921 tgactcgcta cgctcggtcg ttcgactgcg gcgagcggtg tcagctcact caaaagcggt
     1981 aatacggtta tccacagaat caggggataa agccggaaag aacatgtgag caaaaagcaa
     2041 agcaccggaa gaagccaacg ccgcaggcgt ttttccatag gctccgcccc cctgacgagc
     2101 atcacaaaaa tcgacgctca agccagaggt ggcgaaaccc gacaggacta taaagatacc
     2161 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2221 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgtt
     2281 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2341 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2401 acgacttatc gccactggca gcagccattg gtaactgatt tagaggactt tgtcttgaag
     2461 ttatgcacct gttaaggcta aactgaaaga acagattttg gtgagtgcgg tcctccaacc
     2521 cacttacctt ggttcaaaga gttggtagct cagcgaacct tgagaaaacc accgttggta
     2581 gcggtggttt ttctttattt atgagatgat gaatcaatcg gtctatcaag tcaacgaaca
     2641 gctattccgt tactctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc
     2701 agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc caccggaagg
     2761 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
     2821 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2881 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
     2941 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
     3001 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
     3061 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
     3121 accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
     3181 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
     3241 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
     3301 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
     3361 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
     3421 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
     3481 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
     3541 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
     3601 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
     3661 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
     3721 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
     3781 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
     3841 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
     3901 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
     3961 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg
     4021 ctctccctta tgcgactcct gcattaggaa attaatacga ctcactata
//
