LOCUS       sequence-172859-        4749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of anti-CRISPR protein AcrIIA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4749)
  AUTHORS   Rauch BJ, Silvis MR, Hultquist JF, Waters CS, McGregor MJ, Krogan 
            NJ, Bondy-Denomy J
  TITLE     Inhibition of CRISPR-Cas9 with Bacteriophage Proteins.
  JOURNAL   Cell. 2016 Dec 22. pii: S0092-8674(16)31683-X. doi: 
            10.1016/j.cell.2016.12.009.
  PUBMED    28041849
REFERENCE   2  (bases 1 to 4749)
  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: "Cell. 2016 
            Dec 22. pii: S0092-8674(16)31683-X. doi: 
            10.1016/j.cell.2016.12.009."
COMMENT     Sequence Label: pCSW65
FEATURES             Location/Qualifiers
     source          1..4749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        19..303
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     226..245
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      795..881
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      973..1000
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1019..1110
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1111..1179,1180..1971)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1329..1348)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2013..2468
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2100..2119)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2310..2331
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2579..3167
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3068..3087
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3321..3338
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3504..3526
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3664..3683
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3863..4741)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 tactcccgcc attcagagaa gaaaccaatt gtccatattg catcagacat tgccgtcact
       61 gcgtctttta ctggctcttc tcgctaacca aaccggtaac cccgcttatt aaaagcattc
      121 tgtaacaaag cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc tataatcacg
      181 gcagaaaagt ccacattgat tatttgcacg gcgtcacact ttgctatgcc atagcatttt
      241 tatccataag attagcggat cctacctgac gctttttatc gcaactctct actgtttctc
      301 catacccgtt tttttgggct agcaggagga attcaccatg tttaataagg ctgaaattat
      361 gaaacaagct tggaattggt tcaacgatag caacatctgg ttaagcgata ttgaatgggt
      421 aagttacaca gacaaagaaa aatctttctc cgtttgttta aaagctgctt ggtctaaagc
      481 gaaagaagaa gttgaagaat ctaaaaaaga aagcaaacat attgctaaaa gcgaagagtt
      541 gaaagcttgg aattgggctg aacgaaaatt aggcttacac tttaatattt cagatgatga
      601 aaaattcact agtgtaaaag atgaaactaa aataaatttc ggcttaagtg tctgggcttg
      661 tgcaatgaaa gcagttaaac tacacaatga cttgtttcca caaacagcag cttaaaagaa
      721 ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg
      781 gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
      841 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
      901 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
      961 atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt
     1021 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1081 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     1141 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     1201 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     1261 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1321 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     1381 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1441 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     1501 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     1561 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     1621 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     1681 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     1741 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     1801 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     1861 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     1921 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     1981 caagtttact catatatact ttagattgat ttacgcgccc tgtagcggcg cattaagcgc
     2041 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2101 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2161 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2221 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2281 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2341 caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2401 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     2461 tacaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     2521 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2581 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2641 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2701 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     2761 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2821 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2881 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2941 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3001 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3061 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3121 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3181 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3241 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3301 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     3361 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catagggtca tggctgcgcc
     3421 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3481 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3541 accgaaacgc gcgaggcaga aggagatggc gcccaacagt cccccggcca cggggcctgc
     3601 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     3661 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     3721 cacgatgcgt ccggcgtaga ggatctgctc atgtttgaca gcttatcatc gatgcataat
     3781 gtgcctgtca aatggacgaa gcagggattc tgcaaaccct atgctactcc gtcaagccgt
     3841 caattgtctg attcgttacc aattatgaca acttgacggc tacatcattc actttttctt
     3901 cacaaccggc acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga
     3961 aatagagttg atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg
     4021 tgctcaaaag cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa
     4081 tccctaactg ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg
     4141 cgacgctggc gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct
     4201 cgcgtacccg attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca
     4261 gtaacaattg ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc
     4321 cggcgttaat gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc
     4381 gaaagaaccc cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc
     4441 gcggacgaaa gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt
     4501 gatgaatctc tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc
     4561 cctgattttt caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc
     4621 ccagcggtcg gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg
     4681 ccaccagatg ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca
     4741 tacttttca
//
