LOCUS       sequence-469104-        2605 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 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: pHK352.3
FEATURES             Location/Qualifiers
     source          1..2605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      99..185
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      222..971
                     /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     702..721
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     944..961
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1020..1041)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1232..1251
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1464..1486
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1478..1495
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1479..1495
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1496..1530
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     protein_bind    1546..1565
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     protein_bind    1672..1691
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     primer_bind     complement(1795..1813)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1809..1856
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        join(1888..1893,1894..1910,1911..1916)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(2105..2605,1..33)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 cactttgata tcgacccaag taccgccacc taacaattcg ttcaagccga gatcggcttc
       61 ccggccgcgg agttgttcgg taaattggac gtcggcatca aataaaacga aaggctcagt
      121 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
      181 caaatccgcc gccctagagt ggatatgtgg acgatggccg caacggaata gctgttcgtt
      241 gacttgatag accgattgat tcatcatctc ataaataaag aaaaaccacc gctaccaacg
      301 gtggttttct caaggttcgc tgagctacca actctttgaa ccaaggtaag tgggttggag
      361 gaccgcactc accaaaatct gttctttcag tttagcctta acaggtgcat aacttcaaga
      421 caaagtcctc taaatcagtt accaatggct gctgccagtg gcgataagtc gtgtcttacc
      481 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
      541 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata ccaacagcgt
      601 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
      661 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
      721 tatagtcctg tcgggtttcg ccacctctgg cttgagcgtc gatttttgtg atgctcgtca
      781 ggggggcgga gcctatggaa aaacgcctgc ggcgttggct tcttccggtg ctttgctttt
      841 tgctcacatg ttctttccgg ctttatcccc tgattctgtg gataaccgta ttaccgcttt
      901 tgagtgagct gacaccgctc gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga
      961 ggaagcggaa gctggacgat ggcgagccgg cccgagatct cgatcccgcg aaatttgttc
     1021 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     1081 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     1141 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     1201 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     1261 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     1321 cgccgctaca gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     1381 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     1441 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagttttac
     1501 ggctagctca gtcctaggta caatgctagc cagcctgcgg tccgggttca ctgccgtata
     1561 ggcagtgact gagctagtgt actctgtttc agagctatgc tggaaacagc atagcaagtt
     1621 gaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cgttcactgc
     1681 cgtataggca gtcgctggga cgcccgattc tgcagatatc catcacactg gcggccgctc
     1741 gagcagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
     1801 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
     1861 aaggaggaac tatatccgga tgacgtcttg acataagcct gttcggttcg taaactgtaa
     1921 tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt
     1981 aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttgtaca gcctatgcct
     2041 cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc
     2101 aacgatgtta cgcagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt
     2161 aggtggctca agtatgggca tcattcgcac atgtaggctc ggccctgacc aagtcaaatc
     2221 catgcgggct gctcttgatc ttttcggtcg tgagttcgga gacgtagcca cctactccca
     2281 acatcagccg gactccgatt acctcgggaa cttgctccgt agtaagacat tcatcgcgct
     2341 tgctgccttc gaccaagaag cggttgttgg cgctctcgcg gcttacgttc tgcccaagtt
     2401 tgagcagccg cgtagtgaga tctatatcta tgatctcgca gtctccggag agcaccggag
     2461 gcagggcatt gccaccgcgc tcatcaatct cctcaagcat gaggccaacg cgcttggtgc
     2521 ttatgtgatc tacgtgcaag cagattacgg tgacgatccc gcagtggctc tctatacaaa
     2581 gttgggcata cgggaagaag tgatg
//
