LOCUS       sequence-377474-        2268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sgRNA RR1 (target mRFP CDS) on ColE1-AmpR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2268)
  AUTHORS   Kiattisewee C, Karanjia AV, Legut M, Daniloski Z, Koplik SE, Nelson 
            J, Kleinstiver BP, Sanjana NE, Carothers JM, Zalatan JG
  TITLE     Expanding the Scope of Bacterial CRISPR Activation with PAM-Flexible
            dCas9 Variants.
  JOURNAL   ACS Synth Biol. 2022 Nov 15. doi: 10.1021/acssynbio.2c00405.
  PUBMED    36378874
REFERENCE   2  (bases 1 to 2268)
  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: "ACS Synth 
            Biol. 2022 Nov 15. doi: 10.1021/acssynbio.2c00405."
COMMENT     Sequence Label: pCK411.RR1
FEATURES             Location/Qualifiers
     source          1..2268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     132..151
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(370..474)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     542..560
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(564..607)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        625..659
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        680..755
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(847..864)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1018..1606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1098..1117)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1777..2268,1..300,301..369))
                     /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"
ORIGIN
        1 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
       61 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
      121 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
      181 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
      241 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
      301 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
      361 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
      421 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
      481 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
      541 aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt tcgctaagga
      601 tgatttctgg aattctaaag atctttgaca gctagctcag tcctaggtat aatactagta
      661 actttcagtt tagcggtctg ttttagagct agaaatagca agttaaaata aggctagtcc
      721 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttaacgcatg agaaagcccc
      781 cggaagatca ccttccgggg gcttttttat tgcgccttaa gctagaccag cttttgttcc
      841 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
      901 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
      961 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1021 ccacaggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1081 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1141 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1201 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1261 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1321 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1381 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1441 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1501 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1561 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1621 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1681 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     1741 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     1801 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     1861 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     1921 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     1981 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2041 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     2101 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     2161 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     2221 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggc
//
