LOCUS       sequence-172847-        4835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of anti-CRISPR protein AcrIIA1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4835)
  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 4835)
  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: pRAU168
FEATURES             Location/Qualifiers
     source          1..4835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        17..301
                     /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     224..243
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      879..965
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1057..1084
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1103..1194
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1195..1263,1264..2055)
                     /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(1413..1432)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2097..2552
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2184..2203)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2394..2415
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2663..3251
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3152..3171
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3405..3422
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3588..3610
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3748..3767
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3947..4825)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg ccgtcactgc
       61 gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa aagcattctg
      121 taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta taatcacggc
      181 agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat agcattttta
      241 tccataagat tagcggatcc tacctgacgc tttttatcgc aactctctac tgtttctcca
      301 tacccgtttt tttgggctag caggaggaat tcaccatgac tattaaatta ttagatgaat
      361 tcttaaaaaa gcatgatttg acgaggtatc agttaagcaa attaactggt atctcacaaa
      421 acactttaaa agaccagaac gaaaaaccgt taaataaata tactgtttca atactacgct
      481 cgctatcgtt gatttcgggt ttatctgtat cagatgtttt gtttgaatta gaagacatag
      541 aaaaaaattc tgacgatctt gcaggattta aacacctgtt agacaagtac aaactctcat
      601 ttcctgcaca agaattcgaa ttatactgct taatcaaaga gtttgagtct gctaatattg
      661 aagtacttcc ttttacgttc aatagatttg aaaacgaaga acatgtaaat ataaaaaaag
      721 atgtttgtaa agcgttagaa aatgctatca ccgtgttaaa agagaagaaa aatgagttac
      781 tttgataagt ttaaaaggaa ggaactcaga agtgaaacgc cgtagcgccg atggtagtgt
      841 ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
      901 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
      961 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
     1021 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
     1081 tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg
     1141 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1201 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1261 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1321 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1381 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     1441 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1501 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1561 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1621 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1681 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1741 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1801 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1861 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1921 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1981 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2041 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttacgc
     2101 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2161 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2221 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2281 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     2341 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2401 cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg
     2461 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2521 gaattttaac aaaatattaa cgtttacaat ttaaaaggat ctaggtgaag atcctttttg
     2581 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2641 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2701 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2761 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     2821 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2881 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2941 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3001 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3061 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3121 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3181 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3241 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3301 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3361 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3421 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3481 accgcatagg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     3541 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     3601 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagaaggaga tggcgcccaa
     3661 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     3721 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     3781 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatct gctcatgttt
     3841 gacagcttat catcgatgca taatgtgcct gtcaaatgga cgaagcaggg attctgcaaa
     3901 ccctatgcta ctccgtcaag ccgtcaattg tctgattcgt taccaattat gacaacttga
     3961 cggctacatc attcactttt tcttcacaac cggcacggaa ctcgctcggg ctggccccgg
     4021 tgcatttttt aaatacccgc gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga
     4081 cggtggcgat aggcatccgg gtggtgctca aaagcagctt cgcctggctg atacgttggt
     4141 cctcgcgcca gcttaagacg ctaatcccta actgctggcg gaaaagatgt gacagacgcg
     4201 acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt
     4261 gatcgctgat gtactgacaa gcctcgcgta cccgattatc catcggtgga tggagcgact
     4321 cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag cagatttatc gccagcagct
     4381 ccgaatagcg cccttcccct tgcccggcgt taatgatttg cccaaacagg tcgctgaaat
     4441 gcggctggtg cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt gacggccagt
     4501 taagccattc atgccagtag gcgcgcggac gaaagtaaac ccactggtga taccattcgc
     4561 gagcctccgg atgacgaccg tagtgatgaa tctctcctgg cgggaacagc aaaatatcac
     4621 ccggtcggca aacaaattct cgtccctgat ttttcaccac cccctgaccg cgaatggtga
     4681 gattgagaat ataacctttc attcccagcg gtcggtcgat aaaaaaatcg agataaccgt
     4741 tggcctcaat cggcgttaaa cccgccacca gatgggcatt aaacgagtat cccggcagca
     4801 ggggatcatt ttgcgcttca gccatacttt tcata
//
