LOCUS       sequence-170100-        4367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Type II-C anti-CRISPR protein AcrIIC1Boe 
            from Brackiella oedipodis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4367)
  AUTHORS   Pawluk A, Amrani N, Zhang Y, Garcia B, Hidalgo-Reyes Y, Lee J, 
            Edraki A, Shah M, Sontheimer EJ, Maxwell KL, Davidson AR
  TITLE     Naturally Occurring Off-Switches for CRISPR-Cas9.
  JOURNAL   Cell. 2016 Dec 15;167(7):1829-1838.e9. doi: 
            10.1016/j.cell.2016.11.017. Epub 2016 Dec 8.
  PUBMED    27984730
REFERENCE   2  (bases 1 to 4367)
  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.cell.2016.11.017"; journalName: "Cell"; date: 
            "2016-12-15- 15"; volume: "167"; issue: "7"; pages: "1829-1838.e9"
COMMENT     Sequence Label: pEJS430-pCSDest2-AcrIIC1Boe
FEATURES             Location/Qualifiers
     source          1..4367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     60..77
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    194..215
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(230..235,236..253,254..260)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    268..284
                     /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)."
     primer_bind     273..295
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     292..308
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     292..308
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     327..347
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        329..347
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    460..594
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(497..516)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     551..570
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(960..978)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(961..978)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(1013..1997)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     rep_origin      2068..2523
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2155..2174)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2365..2386
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2549..2653
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2654..2722,2723..3514)
                     /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     complement(2872..2891)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3685..4273
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4174..4193
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
       61 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
      121 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
      181 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
      241 tgctcccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
      301 gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg
      361 gtaccgggcc caatgcattg gcgccgcggc cgcgaattaa aaaacctccc acacctcccc
      421 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
      481 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
      541 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctacgta
      601 atacgactca ctatagttct agaggctcga gctacattac attgaattcc caacacgagc
      661 ataagccgaa ggtaaattcg cccatttctt cataacgata ggcgacttct ttgcacgctt
      721 gagaagccac aacatcgtaa tagatgattt caggttcaag gccatcgaag ttttcatcat
      781 attcttcaat accaaggtca cgaacgtagg tgagatcgat gatctctccg tctttgatgc
      841 acgccagtgc ccagccgcaa cctttataag tcactagctc tggttttaat ttaaatacct
      901 ctttcatggt ggcggcttcg aatcgatggg atcctgcaaa aagaacaagt agcttgtatt
      961 ctatagtgtc acctaaatca agcttgctcc gacgtcccca ggcagaatgg cggttcccta
     1021 aacgagcatt gcttatatag acctcccatt aggcacgcct accgcccatt tacgtcaatg
     1081 gaacgcccat ttgcgtcatt gcccctcccc attgacgtca atggggatgt acttggcagc
     1141 catcgcgggc catttaccgc cattgacgtc aatgggagta ctgccaatgt accctggcgt
     1201 acttccaata gtaatgtact tgccaagtta ctattaatag atattgatgt actgccaagt
     1261 gggccattta ccgtcattga cgtcaatagg gggcgtgaga acggatatga atgggcaatg
     1321 agccatccca ttgacgtcaa tggtgggtgg tcctattgac gtcaatgggc attgagccag
     1381 gcgggccatt taccgtaatt gacgtcaatg ggggaggcgc catatacgtc aataggaccg
     1441 cccatatgac gtcaatagga aagaccatat atagagacca ttgacgtcaa tgggggagtg
     1501 gctatgggcg gtattaggaa gccccatata tggtatatgg gaccgcccat tgggaggggc
     1561 tatctacgtc aataggaaaa cccatatatg gaatactata tggcataggg ccaatacata
     1621 gtattgaacc tggccaatag ccatattggc atagggccat attggatatt gcctatatat
     1681 tgatcctggc atatagccaa tatggccgcc attattggca ccatgccaat tcaatatggc
     1741 ggacttggca cagggccaag acccctcctc aaacttggca ccgtgccaag tagacccctc
     1801 cccagttggc acagtgccag gtccgccata ttgaattggc atggtgccaa gtccgccata
     1861 ttgaattggc tggggtccag atccaccata ttgaattggc acgagtccat atacgccata
     1921 ttgaattggc acaggtccag atccaccata ttgaattggc atgagtccat atacgccata
     1981 ttgaattggc ggtcgactgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     2041 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     2101 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2161 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2221 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2281 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2341 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2401 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     2461 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     2521 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2581 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2641 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     2701 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     2761 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     2821 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     2881 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     2941 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3001 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3061 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3121 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3181 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3241 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3301 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3361 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3421 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3481 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     3541 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3601 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3661 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3721 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3781 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3841 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3901 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3961 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4021 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4081 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4141 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4201 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4261 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4321 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggata
//
