LOCUS       sequence-172855-        4744 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 4744)
  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 4744)
  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: pCSW24
FEATURES             Location/Qualifiers
     source          1..4744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..300
                     /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     223..242
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      787..873
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      965..992
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1011..1102
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1103..1171,1172..1963)
                     /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(1321..1340)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2005..2460
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2092..2111)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2302..2323
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2571..3159
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3060..3079
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3313..3330
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3496..3518
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3656..3675
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3855..4733)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 tcccgccatt cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg
       61 tcttttactg gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt
      121 aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca
      181 gaaaagtcca cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat
      241 ccataagatt agcggatcct acctgacgct ttttatcgca actctctact gtttctccat
      301 acccgttttt ttgggctagc aggaggaatt caccatgact aaatataaca aatcagaaat
      361 tatgaaaaac gcttgggcaa tgtttaacag ctacgaatgg gatgttgaaa attttaaatt
      421 tgtatcagca gaaaataaaa cattttcaaa ttgcctaaaa gaagcgtggg cagaagaaaa
      481 agaatatgtt gaacgcaaag caaaagaaac ggcagaagct ccaagatctg aagaagcaaa
      541 agcttgggac tgggcttgtc gtaaattaaa tgtcaatgat ttacaaaaca ttgacgcaac
      601 agacaaagta ttttacgttg ttgacatgca aaaagaaatg tggacttcca acgtctgggc
      661 acaagctatc aaagctgtgg aactttatgt gaaattaggt ttggcataag aactcagaag
      721 tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
      781 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
      841 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
      901 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt
      961 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt
     1021 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     1081 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     1141 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     1201 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     1261 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     1321 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     1381 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     1441 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     1501 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     1561 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     1621 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     1681 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     1741 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     1801 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     1861 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     1921 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     1981 ctcatatata ctttagattg atttacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2041 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2101 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2161 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2221 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2281 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2341 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2401 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     2461 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2521 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2581 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2641 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2701 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     2761 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2821 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2881 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2941 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3001 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3061 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3121 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3181 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3241 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3301 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     3361 ctccttacgc atctgtgcgg tatttcacac cgcatagggt catggctgcg ccccgacacc
     3421 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     3481 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     3541 gcgcgaggca gaaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     3601 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     3661 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     3721 gtccggcgta gaggatctgc tcatgtttga cagcttatca tcgatgcata atgtgcctgt
     3781 caaatggacg aagcagggat tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc
     3841 tgattcgtta ccaattatga caacttgacg gctacatcat tcactttttc ttcacaaccg
     3901 gcacggaact cgctcgggct ggccccggtg cattttttaa atacccgcga gaaatagagt
     3961 tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa
     4021 agcagcttcg cctggctgat acgttggtcc tcgcgccagc ttaagacgct aatccctaac
     4081 tgctggcgga aaagatgtga cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg
     4141 gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc
     4201 cgattatcca tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat
     4261 tgctcaagca gatttatcgc cagcagctcc gaatagcgcc cttccccttg cccggcgtta
     4321 atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac
     4381 cccgtattgg caaatattga cggccagtta agccattcat gccagtaggc gcgcggacga
     4441 aagtaaaccc actggtgata ccattcgcga gcctccggat gacgaccgta gtgatgaatc
     4501 tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg tccctgattt
     4561 ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat aacctttcat tcccagcggt
     4621 cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga
     4681 tgggcattaa acgagtatcc cggcagcagg ggatcatttt gcgcttcagc catacttttc
     4741 atac
//
