LOCUS       sequence-225081-        4484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses type II-C anti-CRISPR protein from S. muelleri in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4484)
  AUTHORS   Lee J, Mir A, Edraki A, Garcia B, Amrani N, Lou HE, Gainetdinov I, 
            Pawluk A, Ibraheim R, Gao XD, Liu P, Davidson AR, Maxwell KL, 
            Sontheimer EJ
  TITLE     Potent Cas9 Inhibition in Bacterial and Human Cells by AcrIIC4 and 
            AcrIIC5 Anti-CRISPR Proteins.
  JOURNAL   MBio. 2018 Dec 4;9(6). pii: mBio.02321-18. doi: 
            10.1128/mBio.02321-18.
  PUBMED    30514786
REFERENCE   2  (bases 1 to 4484)
  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: "MBio."; 
            date: "2018-12-4"; volume: "9(6). pii: mBio.02321-18. doi"; pages: "
            10.1128/mBio.02321-18"
COMMENT     Sequence Label: pEJS542-pCSDest2-AcrIIC5Smu
FEATURES             Location/Qualifiers
     source          1..4484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        76..1060
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1095..1113
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1095..1112
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    1596..1730
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1620..1639)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1674..1693
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1843..1863)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1843..1861)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1882..1898)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1882..1898)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1895..1917)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1906..1922
                     /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)."
     promoter        complement(join(1930..1936,1937..1954,1955..1960))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1975..1996
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2113..2130)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2284..2872)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2364..2383)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3043..3834,3835..3903))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3666..3685
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3904..4008)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(4034..4484,1..5))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4171..4192)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4383..4402
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
       61 ttcgctatta cgccagtcga ccgccaattc aatatggcgt atatggactc atgccaattc
      121 aatatggtgg atctggacct gtgccaattc aatatggcgt atatggactc gtgccaattc
      181 aatatggtgg atctggaccc cagccaattc aatatggcgg acttggcacc atgccaattc
      241 aatatggcgg acctggcact gtgccaactg gggaggggtc tacttggcac ggtgccaagt
      301 ttgaggaggg gtcttggccc tgtgccaagt ccgccatatt gaattggcat ggtgccaata
      361 atggcggcca tattggctat atgccaggat caatatatag gcaatatcca atatggccct
      421 atgccaatat ggctattggc caggttcaat actatgtatt ggccctatgc catatagtat
      481 tccatatatg ggttttccta ttgacgtaga tagcccctcc caatgggcgg tcccatatac
      541 catatatggg gcttcctaat accgcccata gccactcccc cattgacgtc aatggtctct
      601 atatatggtc tttcctattg acgtcatatg ggcggtccta ttgacgtata tggcgcctcc
      661 cccattgacg tcaattacgg taaatggccc gcctggctca atgcccattg acgtcaatag
      721 gaccacccac cattgacgtc aatgggatgg ctcattgccc attcatatcc gttctcacgc
      781 cccctattga cgtcaatgac ggtaaatggc ccacttggca gtacatcaat atctattaat
      841 agtaacttgg caagtacatt actattggaa gtacgccagg gtacattggc agtactccca
      901 ttgacgtcaa tggcggtaaa tggcccgcga tggctgccaa gtacatcccc attgacgtca
      961 atggggaggg gcaatgacgc aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc
     1021 ctaatgggag gtctatataa gcaatgctcg tttagggaac cgccattctg cctggggacg
     1081 tcggagcaag cttgatttag gtgacactat agaatacaag ctacttgttc tttttgcagg
     1141 atcccatcga ttcgaagccg ccaccatgaa caacagcatc aagttccacg tgagctacga
     1201 cggtaccgcg cgtgcgctgt ttaacaccaa ggagcaggcg gaaaaatact gcctggttga
     1261 ggaaattaac gatgagatga acggctataa gcgtaaaagc tgggaggaaa agctgcgtga
     1321 ggaaaactgc gcgagcgtgc aggactgggt tgagaagaac tacaccagca gctatagcga
     1381 cctgttcaac atctgcgaga ttgaagtgag cagcgcgggt caactggtta agatcgacaa
     1441 caccgaggtg gacgatttcg ttgaaaactg ctatggcttt accctggagg acgatctgga
     1501 ggaattcaac aaggcgaaac agtacctgca aaaattttat gcggagtgcg aaaactgact
     1561 cgagcctcta gaactatagt gagtcgtatt acgtagatcc agacatgata agatacattg
     1621 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1681 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1741 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taattcgcgg
     1801 ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt tccctttagt gagggttaat
     1861 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     1921 aattccacac aacatacgag ccgggagcat aaagtgtaaa gcctggggtg cctaatgagt
     1981 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2041 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2101 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2161 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2221 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2281 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2341 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2401 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2461 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2521 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2581 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2641 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2701 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2761 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2821 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2881 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2941 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3001 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3061 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3121 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3181 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3241 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3301 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3361 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3421 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3481 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3541 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3601 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3661 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3721 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3781 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3841 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3901 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3961 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4021 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     4081 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     4141 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     4201 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     4261 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     4321 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     4381 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     4441 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac aggg
//
