LOCUS       sequence-172853-        4643 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of anti-CRISPR protein AcrIIA4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4643)
  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 4643)
  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: pCSW21
FEATURES             Location/Qualifiers
     source          1..4643
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..292
                     /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     215..234
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      678..764
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      856..883
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        902..993
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(994..1062,1063..1854)
                     /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(1212..1231)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1896..2351
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1983..2002)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2193..2214
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2462..3050
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2951..2970
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3204..3221
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3387..3409
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3547..3566
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3746..4624)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 ttcagagaag aaaccaattg tccatattgc atcagacatt gccgtcactg cgtcttttac
       61 tggctcttct cgctaaccaa accggtaacc ccgcttatta aaagcattct gtaacaaagc
      121 gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg cagaaaagtc
      181 cacattgatt atttgcacgg cgtcacactt tgctatgcca tagcattttt atccataaga
      241 ttagcggatc ctacctgacg ctttttatcg caactctcta ctgtttctcc atacccgttt
      301 ttttgggcta gcaggaggaa ttcaccatga atattaatga cttaattaga gaaatcaaaa
      361 acaaagatta cacagtgaaa ttgagtggta cggatagcaa tagtatcaca cagctaatta
      421 ttcgcgttaa taatgatggc aacgagtatg taatttctga aagtgaaaat gaatcaatcg
      481 ttgaaaaatt catctctgca ttcaaaaacg gttggaatca agaatacgag gatgaagaag
      541 aattttataa tgacatgcaa acaatcacct taaaaagtga gttgaactaa aaatatgtca
      601 gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt
      661 agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
      721 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
      781 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
      841 ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc
      901 ttttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
      961 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     1021 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     1081 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     1141 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     1201 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     1261 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     1321 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     1381 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     1441 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     1501 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     1561 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     1621 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     1681 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     1741 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     1801 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     1861 gaccaagttt actcatatat actttagatt gatttacgcg ccctgtagcg gcgcattaag
     1921 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     1981 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2041 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2101 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2161 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2221 actcaaccct atctcgggct attcttttga tttataaggg attttgccga tttcggccta
     2281 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     2341 gtttacaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     2401 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2461 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2521 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2581 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     2641 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2701 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2761 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2821 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2881 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2941 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3001 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3061 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3121 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3181 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     3241 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataggg tcatggctgc
     3301 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3361 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3421 atcaccgaaa cgcgcgaggc agaaggagat ggcgcccaac agtcccccgg ccacggggcc
     3481 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     3541 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     3601 ggccacgatg cgtccggcgt agaggatctg ctcatgtttg acagcttatc atcgatgcat
     3661 aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac tccgtcaagc
     3721 cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca ttcacttttt
     3781 cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta aatacccgcg
     3841 agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata ggcatccggg
     3901 tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag cttaagacgc
     3961 taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag caaacatgct
     4021 gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg tactgacaag
     4081 cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct tccatgcgcc
     4141 gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc ccttcccctt
     4201 gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg
     4261 ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca tgccagtagg
     4321 cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga tgacgaccgt
     4381 agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa acaaattctc
     4441 gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata taacctttca
     4501 ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac
     4561 ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt tgcgcttcag
     4621 ccatactttt catactcccg cca
//
