LOCUS       sequence-500520-        2379 bp    DNA     circular SYN 19-FEB-2026
DEFINITION  R6K plasmid with constituitive red fluorescent protein (mScarlet-I) 
            expression under J23101 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2379)
  AUTHORS   Hina-Nilesh Joshi S, Jenkins C, Ulaeto D, Gorochowski TE
  TITLE     SHARK: A Specialized Host for Assembling R6K Plasmids.
  JOURNAL   ACS Synth Biol. 2026 Jan 13. doi: 10.1021/acssynbio.5c00749.
  PUBMED    41529903
REFERENCE   2  (bases 1 to 2379)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 19, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2026 Jan 13. doi: 10.1021/acssynbio.5c00749."
COMMENT     Sequence Label: pR6K-mScarI
FEATURES             Location/Qualifiers
     source          1..2379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      5..393
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(486..1301)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(486..1301)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1209..1228
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1293..1312
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        1474..1508
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     misc_RNA        1515..1564
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
ORIGIN
        1 gcttgatctg aagatcagca gttcaacctg ttgatagtac gtactaagct ctcatgtttc
       61 acgtactaag ctctcatgtt taacgtacta agctctcatg tttaacgaac taaaccctca
      121 tggctaacgt actaagctct catggctaac gtactaagct ctcatgtttc acgtactaag
      181 ctctcatgtt tgaacaataa aattaatata aatcagcaac ttaaatagcc tctaaggttt
      241 taagttttat aagaaaaaaa agaatatata aggcttttaa agcttttaag gtttaacggt
      301 tgtggacaac aagccaggga tgtaacgcac tgagaagccc ttagagcctc tcaaagcaat
      361 tttgagtgac acaggaacac ttaacggctg acagcaagaa gatcatctta ttaaggggtc
      421 tgacgctcag tggaacgaaa aatcaatcta aagtatatat gagtaaactt ggtctgacag
      481 ttaccttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
      541 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
      601 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
      661 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
      721 acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca
      781 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
      841 gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
      901 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
      961 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     1021 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     1081 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     1141 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     1201 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     1261 cgcggcctgg agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
     1321 tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtac
     1381 atcagagatt ttgagacaca accaattatt gaaggcctcc ctaacggggg gccttttttt
     1441 gtttctggtc tgccgcttga taagtcccta acttttacag ctagctcagt cctaggtatt
     1501 atgctagcct gaagctgtca ccggatgtgc tttccggtct gatgagtccg tgaggacgaa
     1561 acagcctcta caaataattt tgtttaacat aatatttctc ttccggtata aaatggtcag
     1621 taaaggcgaa gcagttatca aagagttcat gcgcttcaaa gttcatatgg aagggtcgat
     1681 gaacgggcac gaatttgaaa ttgaaggcga aggcgaaggc cgcccatatg aagggaccca
     1741 aaccgcaaag cttaaggtta ctaaaggcgg tccattaccc ttttcgtggg acattttaag
     1801 cccacagttt atgtacggga gtcgcgcttt catcaagcac cctgcggaca tcccagatta
     1861 ctacaaacag tctttccccg aggggttcaa gtgggagcgc gtgatgaact tcgaggatgg
     1921 cggagccgtg acggtcaccc aagatacctc tttggaggac ggtacgttga tctacaaagt
     1981 gaaattgcgt ggcacgaatt ttccacctga tgggcctgtc atgcagaaaa agacaatggg
     2041 atgggaagct tccacggagc gcctttaccc agaggacggt gttcttaaag gggatatcaa
     2101 aatggcgctg cgtcttaaag atggaggccg ctacctggcg gacttcaaga ctacttacaa
     2161 ggccaaaaaa ccagtgcaga tgccgggtgc gtacaatgta gatcgtaaat tagatattac
     2221 aagtcacaat gaagattaca cggtcgtaga gcagtatgag cgcagtgagg ggcgtcactc
     2281 tacgggcggt atggacgagt tatacaagta aaggtactcg gtaccaaatt ccagaaaaga
     2341 ggcctcccga aaggggggcc ttttttcgtt ttggtccaa
//
