LOCUS       sequence-323294-        4436 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PJ23110-RBS-PhCFAT-mRuby2-tL3S2P24_KanR_p15A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4436)
  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 4436)
  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: pLNL1141
FEATURES             Location/Qualifiers
     source          1..4436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(243..1058)
                     /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(243..1058)
                     /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     966..985
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1050..1069
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1336..1353)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1470..2015)
                     /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(1550..1569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2240..2274
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     CDS             3670..4377
                     /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 ttttggtccg ctggaaatct gctcgtcagt ggtgctcaca ctgacgaatc atgtacagat
       61 cataccgatg actgcctggc gactcacaac taagcaagac agccggaacc agcgccggcg
      121 aacaccactg catatatggc atatcacaac agtccacgtc tcaagcagtt acagagatgt
      181 tacgaaccac tagtgcactg cagtacagcg gccgcgtgtt acaaccaatt aaccaattct
      241 gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
      301 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
      361 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
      421 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
      481 actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc
      541 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
      601 tgcgcctgag cgaggcgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
      661 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
      721 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
      781 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
      841 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
      901 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
      961 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     1021 ggcctggagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     1081 atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat
     1141 cagagatttt gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag
     1201 gatcagtagc ggagtgtata ctggcttact atgttggcac tgatgagggt gtcagtgaag
     1261 tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt cagcagaata tgtgatacag
     1321 gatatattcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc
     1381 ggaaatggct tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag
     1441 ggaagtgaga gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat
     1501 cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag
     1561 gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt
     1621 gtcattccgc tgttatggcc gcgtttgtct cattccacgc ctgacactca gttccgggta
     1681 ggcagttcgc tccaagctgg actgtatgca cgaacccccc gttcagtccg accgctgcgc
     1741 cttatccggt aactatcgtc ttgagtccaa cccggaaaga catgcaaaag caccactggc
     1801 agcagccact ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct
     1861 aaactgaaag gacaagtttt ggtgactgcg ctcctccaag ccagttacct cggttcaaag
     1921 agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag
     1981 agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt aatcagataa
     2041 aatatttgcg gccgcccctg aattcgcatc tagactgatg agacgtggta gagccacaaa
     2101 cagccggtac aagcaacgat ctccaggacc atctgaatca tgcgcggatg acacgaactc
     2161 acgacggcga tcacagacat taacccacag tacagacact gcgacaacgt ggcaattcgt
     2221 cgcaatacaa cggatttact ttacggctag ctcagtccta ggtacaatgc tagcgctagc
     2281 ttcacacagg aaaccagatc tatgggtaac accgacttcc atgttaccgt gaaaaagaaa
     2341 gaagttgttg cggctgtgtt gcccatgcat catgagcact ggttaccgat gtccaacctg
     2401 gacctgctgc ttccgccgct ggattttggc gtttttttct gctacaaacg ttcgaaaatt
     2461 aacaatgata ccaaagacga cgacgaaacc atcaagaagg cgctggcgga aacgttagtg
     2521 agcttttatg ctcttgcggg ggaagtcgtg ttcaactcac ttggtgagcc agagttattg
     2581 tgcaataatc gtggtgtcga tttctttcac gcgtacgcgg atatcgagtt aaataacctg
     2641 gatctgtatc atccagacgt gagcgtgcac gaaaaactta tccctattaa aaaacacggg
     2701 gtcctgtcgg tacaggttac gggcttaaag tgtggtggaa tcgtcgtggg ctgtaccttt
     2761 gaccatcgtg tggcggatgc gtatagcgca aatatgttcc tggtcgcctg ggcggctatc
     2821 gcgcgtaaag ataataacat caataccgtt attccaagct tccgtcgttc tctcctgaac
     2881 cctcgtcgtc cgcctcaatt tgatgattca tttattgata gtacctacgt ttttctgtct
     2941 tcgccgccta aacaacctaa tgatgtgctg acttcgcgtg tttactatat taattcccag
     3001 gaaatcaacc tgctgcaaag tcaggcgaca cgcaatgggt ctaagcgcag caaacttgaa
     3061 tgcttctccg ccttcctgtg gaaaaccatc gcggaaggtg gcattgacga tagtaaacgc
     3121 tgcaaactgg ggattgtggt tgatggacgt cagcgtctgc gccatgatag ctcaacgacc
     3181 atgaaaaact atttcggtaa tgttctgagt gtaccttaca ccgaagcgtc cgtaggccag
     3241 ttaaaacaga cccccctggg taaggtggcc gacctggtcc acacctgcct ggataatgta
     3301 gctaatgaac atcacttccc atcactgatc gattgggtgg aacttcaccg tccccgccag
     3361 gctattgtaa aagtctactg taaggatgaa tgtaatgatg aagcggcaat cgtagtttcg
     3421 tcgggtctgc gtttcccact ctctcaggta aactttggct gggggtgtcc ggattttggc
     3481 agctatatct tcccgtgggg tggccaaacc gggtatgtaa tgccaatgcc ttcgccgaat
     3541 aagaatggcg attggatcgt ttatatgcat ctccagaaga agcatttgga cctggtggaa
     3601 acgcgtgccc ctcacatctt ccacccgctg actgcgtgct atttggatct tacagcaacg
     3661 tatggatccg tgtccaaagg agaggagtta atcaaggaaa acatgagaat gaaagttgtc
     3721 atggagggct ccgttaatgg tcaccaattc aagtgtacag gggaaggtga aggtaatcct
     3781 tacatgggta cacaaactat gagaattaaa gtaattgaag gcggaccact accatttgca
     3841 tttgacattc tggcaacgtc attcatgtac ggatcacgaa ctttcatcaa gtaccctaaa
     3901 ggtataccag actttttcaa gcaatctttt ccagagggtt ttacatggga aagggttaca
     3961 agatacgaag atgggggtgt cgtcacagtt atgcaagata cttcattaga agatggctgc
     4021 cttgtctatc atgtgcaagt aagaggggtg aattttcctt ctaacggacc tgtgatgcag
     4081 aaaaagacca aaggttggga accaaatact gaaatgatgt acccagctga tggaggtttg
     4141 agaggctaca cacacatggc gcttaaagtt gatggtggag gtcatttgtc ttgtagtttt
     4201 gttaccactt atcgttctaa aaagactgtt ggcaatatca aaatgccagg aatacatgct
     4261 gtagaccaca gactagaaag actcgaagag agcgataacg aaatgttcgt tgtacagaga
     4321 gagcatgccg tagccaaatt tgctggctta ggcggtggta tggatgaatt gtataagtaa
     4381 ctcgagtggc ctcggtacca aattccagaa aagacacccg aaagggtgtt ttttcg
//
