LOCUS       sequence-414098-        2770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AcrIIC1-mediated inhibition of ThermoCas9 and GeoCas9 cleavage 
            activity in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2770)
  TITLE     van der Oost E. coli CRISPR plasmids
REFERENCE   2  (bases 1 to 2770)
  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"
COMMENT     Sequence Label: pAcr
FEATURES             Location/Qualifiers
     source          1..2770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      121..709
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     610..629
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     863..880
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        997..1115
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     primer_bind     complement(1453..1471)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1467..1514
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1609..1631
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1669..1687)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1755..1859
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1860..1928,1929..2720)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2078..2097)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
       61 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
      121 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
      181 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      241 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      301 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      361 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      421 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      481 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      541 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      601 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      661 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      721 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      781 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      841 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      901 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      961 tcccgactgg aaagcgggca gtgagcgcaa cgcaatcacc acaattcagc aaattgtgaa
     1021 catcatcacg ttcatctttc cctggttgcc aatggcccat tttcctgtca gtaacgagaa
     1081 ggtcgcgaat tcaggcgctt tttagactgg tcgtagggag accacaacgg tttccctcta
     1141 gaaataattt tgtttaacta taagaaggag atatacatat gaataaaaca tataaaatcg
     1201 gaaaaaatgc gggatatgac ggctgcgggt tatgtttagc cgcgatctca gaaaatgaag
     1261 ctattaaagt taagtatctg cgcgatatct gtccagacta tgacggcgac gacaaagctg
     1321 aggactggct gagatgggga acggatagcc gcgtgaaagc tgcggcttta gaaatggagc
     1381 aatatgctta tacgtcggtt gggatggcgt catgttggga gtttgttgaa ctatgaccta
     1441 gagcggccgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     1501 tgaggggttt tttgttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     1561 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     1621 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     1681 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     1741 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     1801 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     1861 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     1921 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     1981 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2041 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2101 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2161 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2221 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2281 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2341 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2401 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     2461 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     2521 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     2581 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     2641 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     2701 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     2761 taaaacttca
//
