LOCUS       sequence-469106-        2605 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 2605)
  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 2605)
  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..2605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      97..183
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      220..969
                     /direction=RIGHT
                     /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     700..719
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     942..959
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1018..1039)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1230..1249
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1462..1484
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1476..1493
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1477..1493
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1494..1528
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     protein_bind    1544..1563
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     protein_bind    1670..1689
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     primer_bind     complement(1793..1811)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1807..1854
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        join(1886..1891,1892..1908,1909..1914)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(2103..2605,1..31)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 ctttgatatc gacccaagta ccgccaccta acaattcgtt caagccgaga tcggcttccc
       61 ggccgcggag ttgttcggta aattggacgt cggcatcaaa taaaacgaaa ggctcagtcg
      121 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
      181 aatccgccgc cctagagtgg atatgtggac gatggccgca acggaatagc tgttcgttga
      241 cttgatagac cgattgattc atcatctcat aaataaagaa aaaccaccgc taccaacggt
      301 ggttttctca aggttcgctg agctaccaac tctttgaacc aaggtaagtg ggttggagga
      361 ccgcactcac caaaatctgt tctttcagtt tagccttaac aggtgcataa cttcaagaca
      421 aagtcctcta aatcagttac caatggctgc tgccagtggc gataagtcgt gtcttaccgg
      481 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
      541 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc aacagcgtga
      601 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      661 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      721 tagtcctgtc gggtttcgcc acctctggct tgagcgtcga tttttgtgat gctcgtcagg
      781 ggggcggagc ctatggaaaa acgcctgcgg cgttggcttc ttccggtgct ttgctttttg
      841 ctcacatgtt ctttccggct ttatcccctg attctgtgga taaccgtatt accgcttttg
      901 agtgagctga caccgctcgc cgcagtcgaa cgaccgagcg tagcgagtca gtgagcgagg
      961 aagcggaagc tggacgatgg cgagccggcc cgagatctcg atcccgcgaa atttgttcca
     1021 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     1081 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     1141 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     1201 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     1261 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     1321 ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     1381 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     1441 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtttgacag
     1501 ctagctcagt cctaggtata atgctagcca gcctgcggtc cgggttcact gccgtatagg
     1561 cagtgactga gctagtgtac tctgtttcag agctatgctg gaaacagcat agcaagttga
     1621 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgcg ttcactgccg
     1681 tataggcagt cgctgggacg cccgattctg cagatatcca tcacactggc ggccgctcga
     1741 gcagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     1801 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     1861 ggaggaacta tatccggatg acgtcttgac ataagcctgt tcggttcgta aactgtaatg
     1921 caagtagcgt atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa
     1981 cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttgtacagc ctatgcctcg
     2041 ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa
     2101 cgatgttacg cagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttag
     2161 gtggctcaag tatgggcatc attcgcacat gtaggctcgg ccctgaccaa gtcaaatcca
     2221 tgcgggctgc tcttgatctt ttcggtcgtg agttcggaga cgtagccacc tactcccaac
     2281 atcagccgga ctccgattac ctcgggaact tgctccgtag taagacattc atcgcgcttg
     2341 ctgccttcga ccaagaagcg gttgttggcg ctctcgcggc ttacgttctg cccaagtttg
     2401 agcagccgcg tagtgagatc tatatctatg atctcgcagt ctccggagag caccggaggc
     2461 agggcattgc caccgcgctc atcaatctcc tcaagcatga ggccaacgcg cttggtgctt
     2521 atgtgatcta cgtgcaagca gattacggtg acgatcccgc agtggctctc tatacaaagt
     2581 tgggcatacg ggaagaagtg atgca
//
