LOCUS       sequence-323303-        4337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PJ23110-RBS-RsVS-mRuby2-tL3S2P24_KanR_p15A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4337)
  AUTHORS   Latimer LN, Russ ZN, Lucas J, Dueber JE
  TITLE     Exploration of Acetylation as a Base-Labile Protecting Group in 
            Escherichia coli for an Indigo Precursor.
  JOURNAL   ACS Synth Biol. 2020 Sep 18. doi: 10.1021/acssynbio.0c00297.
  PUBMED    32886882
REFERENCE   2  (bases 1 to 4337)
  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: "ACS Synth 
            Biol. 2020 Sep 18. doi: 10.1021/acssynbio.0c00297."
COMMENT     Sequence Label: pLNL1143
FEATURES             Location/Qualifiers
     source          1..4337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(234..1049)
                     /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(234..1049)
                     /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     957..976
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1041..1060
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1327..1344)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1461..2006)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1541..1560)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2231..2265
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     CDS             3562..4269
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTHM
                     ALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAVA
                     KFAGLGGGMDELYK"
ORIGIN
        1 gctggaaatc tgctcgtcag tggtgctcac actgacgaat catgtacaga tcataccgat
       61 gactgcctgg cgactcacaa ctaagcaaga cagccggaac cagcgccggc gaacaccact
      121 gcatatatgg catatcacaa cagtccacgt ctcaagcagt tacagagatg ttacgaacca
      181 ctagtgcact gcagtacagc ggccgcgtgt tacaaccaat taaccaattc tgattagaaa
      241 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
      301 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
      361 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
      421 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
      481 ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta
      541 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
      601 gcgaggcgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
      661 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
      721 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
      781 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
      841 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
      901 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
      961 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctggag
     1021 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     1081 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
     1141 tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagtag
     1201 cggagtgtat actggcttac tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg
     1261 tggcaggaga aaaaaggctg caccggtgcg tcagcagaat atgtgataca ggatatattc
     1321 cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc
     1381 ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag
     1441 agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc
     1501 tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1561 cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg
     1621 ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg
     1681 ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg
     1741 taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac
     1801 tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa
     1861 ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag
     1921 ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga
     1981 ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata aaatatttgc
     2041 ggccgcccct gaattcgcat ctagactgat gagacgtggt agagccacaa acagccggta
     2101 caagcaacga tctccaggac catctgaatc atgcgcggat gacacgaact cacgacggcg
     2161 atcacagaca ttaacccaca gtacagacac tgcgacaacg tggcaattcg tcgcaataca
     2221 acggatttac tttacggcta gctcagtcct aggtacaatg ctagcgctag cttcacacag
     2281 gaaaccagat ctatggcacc gcagatggaa aaagtgagcg aggagctgat tctgcctagc
     2341 tccccgaccc cgcagagctt aaaatgctac aaaattagcc atttggatca gctgttgctg
     2401 acatgccaca tcccttttat cctgttctac cctaatccgt tagatagcaa tctggatccg
     2461 gcgcagacga gccaacactt aaaacaatcg ctgtcgaaag tgcttacaca tttttaccca
     2521 ctggcgggac gtattaacgt gaattcttcg gtggattgca atgattctgg cgttccgttt
     2581 gtagaagcgc gtgttcaagc gcaactgtct caggcaatcc agaacgtggt tgagctggaa
     2641 aaattggatc agtacctgcc aagtgctgct taccccggtg gaaaaattga agttaacgaa
     2701 gatgtcccgc tggccgtgaa aatcagtttc tttgaatgtg gaggcaccgc aattggtgtc
     2761 aacctcagcc acaagattgc tgacgtgctg tcgcttgcta cgttcctgaa cgcctggacc
     2821 gcaacctgcc gtggagaaac ggagatcgtc ctgcctaact ttgacctggc cgcccgtcac
     2881 tttccgccgg tagataatac cccgtcaccg gaactcgtcc cggatgaaaa tgtcgtgatg
     2941 aagcgctttg tcttcgacaa agagaaaatc ggcgcgttgc gcgctcaggc gagctctgca
     3001 tccgaagaaa aaaattttag ccgcgtccaa cttgtcgtgg cctatatctg gaaacatgta
     3061 attgatgtga cgcgtgctaa atatggcgca aaaaacaagt ttgtggtagt acaggcagtc
     3121 aacctgcgtt cgcgtatgaa ccccccgctg ccgcactacg cgatgggtaa tattgcgacc
     3181 ctgctgttcg ccgctgtcga cgccgaatgg gataaagatt ttcccgacct cattggtccg
     3241 ttgcgcactt ctcttgaaaa aaccgaagac gatcataatc atgaactttt aaaaggtatg
     3301 acctgtcttt acgaactgga accacaggaa ctgctgagtt ttacaagctg gtgtcgtctg
     3361 gggttttatg atttagattt tggctggggg aaaccgctga gcgcctgtac cacgacattc
     3421 ccgaaacgta acgcggcctt actgatggac acgcgttctg gagatggcgt cgaggcgtgg
     3481 cttccaatgg cggaagatga aatggcgatg ctgccggtgg agctgctgtc tttggtggat
     3541 tccgactttt caaaaggatc cgtgtccaaa ggagaggagt taatcaagga aaacatgaga
     3601 atgaaagttg tcatggaggg ctccgttaat ggtcaccaat tcaagtgtac aggggaaggt
     3661 gaaggtaatc cttacatggg tacacaaact atgagaatta aagtaattga aggcggacca
     3721 ctaccatttg catttgacat tctggcaacg tcattcatgt acggatcacg aactttcatc
     3781 aagtacccta aaggtatacc agactttttc aagcaatctt ttccagaggg ttttacatgg
     3841 gaaagggtta caagatacga agatgggggt gtcgtcacag ttatgcaaga tacttcatta
     3901 gaagatggct gccttgtcta tcatgtgcaa gtaagagggg tgaattttcc ttctaacgga
     3961 cctgtgatgc agaaaaagac caaaggttgg gaaccaaata ctgaaatgat gtacccagct
     4021 gatggaggtt tgagaggcta cacacacatg gcgcttaaag ttgatggtgg aggtcatttg
     4081 tcttgtagtt ttgttaccac ttatcgttct aaaaagactg ttggcaatat caaaatgcca
     4141 ggaatacatg ctgtagacca cagactagaa agactcgaag agagcgataa cgaaatgttc
     4201 gttgtacaga gagagcatgc cgtagccaaa tttgctggct taggcggtgg tatggatgaa
     4261 ttgtataagt aactcgagtg gcctcggtac caaattccag aaaagacacc cgaaagggtg
     4321 ttttttcgtt ttggtcc
//
