LOCUS       sequence-469691-        2607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Message phagemid carrying sgRNA3 (prom. J23110, 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: pHK352.3
FEATURES             Location/Qualifiers
     source          1..2607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..750)
                     /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(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(251..270)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(787..873)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(939..1472)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1661..1666,1667..1683,1684..1689))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     terminator      complement(1721..1768)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1764..1782
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    complement(1888..1907)
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     protein_bind    complement(2014..2033)
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     promoter        complement(2049..2083)
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     primer_bind     complement(2084..2101)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2084..2100)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2093..2115)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2328..2347)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2538..2559
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggtgt
       61 cagctcactc aaaagcggta atacggttat ccacagaatc aggggataaa gccggaaaga
      121 acatgtgagc aaaaagcaaa gcaccggaag aagccaacgc cgcaggcgtt tttccatagg
      181 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gccagaggtg gcgaaacccg
      241 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      301 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      361 tctcatagct cacgctgttg gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      421 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      481 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccattgg taactgattt
      541 agaggacttt gtcttgaagt tatgcacctg ttaaggctaa actgaaagaa cagattttgg
      601 tgagtgcggt cctccaaccc acttaccttg gttcaaagag ttggtagctc agcgaacctt
      661 gagaaaacca ccgttggtag cggtggtttt tctttattta tgagatgatg aatcaatcgg
      721 tctatcaagt caacgaacag ctattccgtt gcggccatcg tccacatatc cactctaggg
      781 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
      841 ccagtctttc gactgagcct ttcgttttat ttgatgccga cgtccaattt accgaacaac
      901 tccgcggccg ggaagccgat ctcggcttga acgaattgtt aggtggcggt acttgggtcg
      961 atatcaaagt gcatcacttc ttcccgtatg cccaactttg tatagagagc cactgcggga
     1021 tcgtcaccgt aatctgcttg cacgtagatc acataagcac caagcgcgtt ggcctcatgc
     1081 ttgaggagat tgatgagcgc ggtggcaatg ccctgcctcc ggtgctctcc ggagactgcg
     1141 agatcataga tatagatctc actacgcggc tgctcaaact tgggcagaac gtaagccgcg
     1201 agagcgccaa caaccgcttc ttggtcgaag gcagcaagcg cgatgaatgt cttactacgg
     1261 agcaagttcc cgaggtaatc ggagtccggc tgatgttggg agtaggtggc tacgtctccg
     1321 aactcacgac cgaaaagatc aagagcagcc cgcatggatt tgacttggtc agggccgagc
     1381 ctacatgtgc gaatgatgcc catacttgag ccacctaact ttgttttagg gcgactgccc
     1441 tgctgcgtaa catcgttgct gctgcgtaac atcgttgctg ctccataaca tcaaacatcg
     1501 acccacggcg taacgcgctt gctgcttgga tgcccgaggc ataggctgta caaaaaaaca
     1561 gtcataacaa gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt
     1621 tctggaccag ttgcgtgagc gcatacgcta cttgcattac agtttacgaa ccgaacaggc
     1681 ttatgtcaag acgtcatccg gatatagttc ctcctttcag caaaaaaccc ctcaagaccc
     1741 gtttagaggc cccaaggggt tatgctagtt attgctcagc ggtggcagca gccaactcag
     1801 cttcctttcg ggctttgtta gcagccggat ctgctcgagc ggccgccagt gtgatggata
     1861 tctgcagaat cgggcgtccc agcgagcctg cctatacggc agtgaacgca ccgactcggt
     1921 gccacttttt caagttgata acggactagc cttatttcaa cttgctatgc tgtttccagc
     1981 atagctctga aacagagtac actagctcag tcactgccta tacggcagtg aacccggacc
     2041 gcaggctggc tagcattgta cctaggactg agctagccgt aaaactggcc gtcgttttac
     2101 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     2161 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     2221 gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2281 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2341 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2401 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2461 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2521 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaaatttcg cgggatcgag
     2581 atctcgggcc ggctcgccat cgtccag
//
