LOCUS       sequence-469669-        3828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Message phagemid carrying sgRNA1 (prom. J23119, backbone pBR322).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3828)
  AUTHORS   Kusumawardhani, H., Zoppi, F., Avendano, R. et al.
  TITLE     Engineering intercellular communication using M13 phagemid and 
            CRISPR-based gene regulation for multicellular computing in 
            Escherichia coli
  JOURNAL   Nat Commun 16, 3569 (2025)
REFERENCE   2  (bases 1 to 3828)
  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: "Nat Commun 
            16, 3569 (2025)"
COMMENT     Sequence Label: pHK002.1
FEATURES             Location/Qualifiers
     source          1..3828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(617..1408,1409..1477))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1240..1259
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1478..1582)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1650..1668
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1713..1760)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1756..1774
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    complement(1880..1899)
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     protein_bind    complement(2006..2025)
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     promoter        complement(2041..2075)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     primer_bind     complement(2076..2093)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2076..2092)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2085..2107)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2320..2339)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2530..2551
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3065..3256
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3250..3272)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3358..3500
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3413..3432
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3515..3532)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(3686..3828,1..446))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3766..3785)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
       61 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
      121 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
      181 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
      241 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
      301 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
      361 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
      421 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
      481 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
      541 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
      601 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
      661 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
      721 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
      781 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
      841 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
      901 ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt
      961 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     1021 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     1081 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     1141 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     1201 tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca
     1261 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     1321 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     1381 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     1441 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     1501 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     1561 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     1621 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc
     1681 aagaattatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
     1741 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
     1801 cgggctttgt tagcagccgg atctgctcga gcggccgcca gtgtgatgga tatctgcaga
     1861 atcgggcgtc ccagcgagcc tgcctatacg gcagtgaacg caccgactcg gtgccacttt
     1921 ttcaagttga taacggacta gccttatttc aacttgctat gctgtttcca gcatagctct
     1981 gaaaccacta gagtctagct tgagactgcc tatacggcag tgaacccgga ccgcaggctg
     2041 gctagcatta tacctaggac tgagctagct gtcaaactgg ccgtcgtttt acaacgtcgt
     2101 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     2161 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     2221 aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2281 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2341 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2401 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2461 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2521 tctttaatag tggactcttg ttccaaactg gaacaaattt cgcgggatcg agatctcggg
     2581 cagcgttggg tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct
     2641 aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg
     2701 aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt
     2761 ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat
     2821 ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg
     2881 ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt
     2941 accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat
     3001 cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc
     3061 atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac
     3121 attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga
     3181 atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga
     3241 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     3301 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     3361 agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca
     3421 gagcagattg tactgagagt gcaccatata tgcggtgtga aataccgcac agatgcgtaa
     3481 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3541 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3601 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3661 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3721 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3781 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgc
//
