LOCUS       sequence-172849-        5668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of anti-CRISPR protein AcrIIA2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5668)
  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 5668)
  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: pRAU171
FEATURES             Location/Qualifiers
     source          1..5668
                     /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      799..885
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      977..1004
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1023..1114
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1711..1797
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1889..1916
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1935..2026
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2027..2095,2096..2887)
                     /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(2245..2264)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2929..3384
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3016..3035)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3226..3247
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3495..4083
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3984..4003
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4237..4254
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4420..4442
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4580..4599
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4779..5657)
                     /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 caccatgaca ttaacaagag cacaaaaaaa
      361 gtatgcagaa gcgatgcacg agtttataaa tatggtggat gactttgaag agtctacacc
      421 ggattttgca aaggaagttc tacatgattc tgactatgta gttattacaa aaaatgaaaa
      481 atatgcagta gctctttgct ctcttagcac tgatgaatgt gaatatgata ctaacttata
      541 cttagatgaa aaattggttg attactcaac agttgacgta aacggtgtga catactacat
      601 caacattgtt gaaactaacg atatcgatga tttagaaatc gctacggatg aagatgagat
      661 gaaaagtggc aaccaagaaa ttattttaaa aagtgagttg aagtaaaaaa tatgactatt
      721 agaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag
      781 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
      841 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
      901 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
      961 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact
     1021 cttttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     1081 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     1141 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     1201 gaaagtctcc atacccgttt ttttgggcta gcaggaggaa ttcaccatga cattaacaag
     1261 agcacaaaaa aagtatgcag aagcgatgca cgagtttata aatatggtgg atgactttga
     1321 agagtctaca ccggattttg caaaggaagt tctacatgat tctgactatg tagttattac
     1381 aaaaaatgaa aaatatgcag tagctctttg ctctcttagc actgatgaat gtgaatatga
     1441 tactaactta tacttagatg aaaaattggt tgattactca acagttgacg taaacggtgt
     1501 gacatactac atcaacattg ttgaaactaa cgatatcgat gatttagaaa tcgctacgga
     1561 tgaagatgag atgaaaagtg gcaaccaaga aattatttta aaaagtgagt tgaagtaaaa
     1621 aatatgacta ttagaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc
     1681 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
     1741 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     1801 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
     1861 ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg
     1921 tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1981 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     2041 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     2101 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     2161 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     2221 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     2281 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     2341 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     2401 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     2461 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     2521 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     2581 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2641 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2701 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     2761 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     2821 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2881 ttggtaactg tcagaccaag tttactcata tatactttag attgatttac gcgccctgta
     2941 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3001 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3061 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3121 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     3181 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3241 aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc
     3301 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3361 acaaaatatt aacgtttaca atttaaaagg atctaggtga agatcctttt tgataatctc
     3421 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3481 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3541 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3601 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     3661 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3721 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3781 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3841 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3901 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3961 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4021 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4081 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4141 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     4201 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     4261 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4321 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4381 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4441 ggttttcacc gtcatcaccg aaacgcgcga ggcagaagga gatggcgccc aacagtcccc
     4501 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     4561 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     4621 cgccggtgat gccggccacg atgcgtccgg cgtagaggat ctgctcatgt ttgacagctt
     4681 atcatcgatg cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc
     4741 tactccgtca agccgtcaat tgtctgattc gttaccaatt atgacaactt gacggctaca
     4801 tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt
     4861 ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg
     4921 ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc
     4981 cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac
     5041 aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg
     5101 atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc
     5161 gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag
     5221 cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg
     5281 tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat
     5341 tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc
     5401 ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg
     5461 caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga
     5521 atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca
     5581 atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca
     5641 ttttgcgctt cagccatact tttcatac
//
