LOCUS       sequence-469676-        2607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Message phagemid carrying sgRNA3 (prom. J23119, backbone RSF1030).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2607)
  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 2607)
  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: pHK052.3
FEATURES             Location/Qualifiers
     source          1..2607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(44..793)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(54..71)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(294..313)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(830..916)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(982..1515)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1704..1709,1710..1726,1727..1732))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     terminator      complement(1764..1811)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1807..1825
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    complement(1931..1950)
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     protein_bind    complement(2057..2076)
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     promoter        complement(2092..2126)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     primer_bind     complement(2127..2144)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2127..2143)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2136..2158)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2371..2390)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2581..2602
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 atttcgcggg atcgagatct cgggccggct cgccatcgtc cagcttccgc ttcctcgctc
       61 actgactcgc tacgctcggt cgttcgactg cggcgagcgg tgtcagctca ctcaaaagcg
      121 gtaatacggt tatccacaga atcaggggat aaagccggaa agaacatgtg agcaaaaagc
      181 aaagcaccgg aagaagccaa cgccgcaggc gtttttccat aggctccgcc cccctgacga
      241 gcatcacaaa aatcgacgct caagccagag gtggcgaaac ccgacaggac tataaagata
      301 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
      361 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
      421 ttggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
      481 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
      541 acacgactta tcgccactgg cagcagccat tggtaactga tttagaggac tttgtcttga
      601 agttatgcac ctgttaaggc taaactgaaa gaacagattt tggtgagtgc ggtcctccaa
      661 cccacttacc ttggttcaaa gagttggtag ctcagcgaac cttgagaaaa ccaccgttgg
      721 tagcggtggt ttttctttat ttatgagatg atgaatcaat cggtctatca agtcaacgaa
      781 cagctattcc gttgcggcca tcgtccacat atccactcta gggcggcgga tttgtcctac
      841 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct ttcgactgag
      901 cctttcgttt tatttgatgc cgacgtccaa tttaccgaac aactccgcgg ccgggaagcc
      961 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac
     1021 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc
     1081 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag
     1141 cgcggtggca atgccctgcc tccggtgctc tccggagact gcgagatcat agatatagat
     1201 ctcactacgc ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc caacaaccgc
     1261 ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta
     1321 atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag
     1381 atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat
     1441 gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt
     1501 gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
     1561 cttgctgctt ggatgcccga ggcataggct gtacaaaaaa acagtcataa caagccatga
     1621 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     1681 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aagacgtcat
     1741 ccggatatag ttcctccttt cagcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     1801 ggttatgcta gttattgctc agcggtggca gcagccaact cagcttcctt tcgggctttg
     1861 ttagcagccg gatctgctcg agcggccgcc agtgtgatgg atatctgcag aatcgggcgt
     1921 cccagcgagc ctgcctatac ggcagtgaac gcaccgactc ggtgccactt tttcaagttg
     1981 ataacggact agccttattt caacttgcta tgctgtttcc agcatagctc tgaaacagag
     2041 tacactagct cagtcactgc ctatacggca gtgaacccgg accgcaggct ggctagcatt
     2101 atacctagga ctgagctagc tgtcaaactg gccgtcgttt tacaacgtcg tgactgggaa
     2161 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     2221 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2281 tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2341 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2401 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2461 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     2521 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2581 gtggactctt gttccaaact ggaacaa
//
