LOCUS       sequence-323030-        4445 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PJ23110-RBS-RhAAT1-mRuby2-tL3S2P24_KanR_p15A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4445)
  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 4445)
  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: pLNL1127
FEATURES             Location/Qualifiers
     source          1..4445
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(211..1026)
                     /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(211..1026)
                     /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     934..953
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1018..1037
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1304..1321)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1438..1983)
                     /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(1518..1537)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2208..2242
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     CDS             3647..4354
                     /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 tgctcacact gacgaatcat gtacagatca taccgatgac tgcctggcga ctcacaacta
       61 agcaagacag ccggaaccag cgccggcgaa caccactgca tatatggcat atcacaacag
      121 tccacgtctc aagcagttac agagatgtta cgaaccacta gtgcactgca gtacagcggc
      181 cgcgtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa
      241 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
      301 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
      361 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
      421 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg
      481 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
      541 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg aggcgaaata cgcgatcgct
      601 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
      661 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc
      721 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
      781 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
      841 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
      901 tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
      961 atcagcatcc atgttggaat ttaatcgcgg cctggagcaa gacgtttccc gttgaatatg
     1021 gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga
     1081 tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttgttg
     1141 aataaatcga acttttgctg agttgaagga tcagtagcgg agtgtatact ggcttactat
     1201 gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac
     1261 cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc
     1321 tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc
     1381 tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt
     1441 ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg
     1501 aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc
     1561 tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca
     1621 ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg
     1681 aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     1741 cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta
     1801 gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct
     1861 cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg
     1921 ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct
     1981 caagaagatc atcttattaa tcagataaaa tatttgcggc cgcccctgaa ttcgcatcta
     2041 gactgatgag acgtggtaga gccacaaaca gccggtacaa gcaacgatct ccaggaccat
     2101 ctgaatcatg cgcggatgac acgaactcac gacggcgatc acagacatta acccacagta
     2161 cagacactgc gacaacgtgg caattcgtcg caatacaacg gatttacttt acggctagct
     2221 cagtcctagg tacaatgcta gcgctagctt cacacaggaa accagatcta tggagaagat
     2281 cgaagtctct ataatttcaa gagacacaat aaaaccttct gctgcctcat cttctttaca
     2341 tccttacaag ttgtcaataa ttgatcaatt cacacctaca acttattttc ctgtcatatt
     2401 cttttaccca attactgaca gagtctttaa tttgccacag acattaaccg acttaaagaa
     2461 taccgtatca caggctttaa ctttatatca tccattatct ggaaggatca agaacaattt
     2521 atacattgat gatttcgaag ctggaatccc ttacttagaa gccagagtca attttcatat
     2581 gatagatttc ttaagattgc ctaaaattga atggttaaat gagtttgttc ctatggctcc
     2641 ttacaggaaa gaaactattt cagagttctt gccattgttg ggtattcaag taaacatatt
     2701 tgattcaggt atcgctattg gagtatcatt ttcacacaaa attaatgacg gacaaaccgc
     2761 atcatgtttc ttaaagtcat gggttgcaat atttaggggt tacagaaaca agattataca
     2821 tccaaatttg tcacaagctg ctttattgtt gccatcaagg gatgatttgc cagaaaagta
     2881 tgtagccatg atggagagaa tgtggttcgg tgaaaagaag gtcgtcacta gaagatttgt
     2941 tttcgatgct aaagcaatat cagcattaca agatgaagga aagtcagaat acgtccctaa
     3001 accttctagg gtacaagcat tgacaggatt cttatggaag catcaattgg cagcctcaag
     3061 agcattgtct tctggaactt ctactaggtt ttctgttgcc tctcagactg tcaatttaag
     3121 atcaaagatg aatatgaaga ctactttaga taacgctatt ggtaatattt tcttgtgggc
     3181 ctcagccaga ttggacttga atgacactgc cccaggatca tctgacttga agttgtgtga
     3241 tttggtaaat ttgttgaatg agtctatcaa agaatttaat tcagactatt tggaaatttt
     3301 gaaaggaaag gaaggttacg gaggtatgtg tgatttattg gatttcatgg aggagggttc
     3361 ttttgttgag ccagctccag aattttattc attctcttca tggaccagat tcttcgatca
     3421 agtcgacttt ggttggggta ggccatcttg ggtaggtttc tctggtagag ttgaaactag
     3481 aaattttact atatttgttg aaacccaatg tgatgatgga atcgatgctt gggttaccgt
     3541 cgacgaaaag caaatggcta tgttagaaca agacccacag ttcttggctt ttgcttcacc
     3601 taatcctaga atatctattg catcatctgt tggtatggac ggatccgtgt ccaaaggaga
     3661 ggagttaatc aaggaaaaca tgagaatgaa agttgtcatg gagggctccg ttaatggtca
     3721 ccaattcaag tgtacagggg aaggtgaagg taatccttac atgggtacac aaactatgag
     3781 aattaaagta attgaaggcg gaccactacc atttgcattt gacattctgg caacgtcatt
     3841 catgtacgga tcacgaactt tcatcaagta ccctaaaggt ataccagact ttttcaagca
     3901 atcttttcca gagggtttta catgggaaag ggttacaaga tacgaagatg ggggtgtcgt
     3961 cacagttatg caagatactt cattagaaga tggctgcctt gtctatcatg tgcaagtaag
     4021 aggggtgaat tttccttcta acggacctgt gatgcagaaa aagaccaaag gttgggaacc
     4081 aaatactgaa atgatgtacc cagctgatgg aggtttgaga ggctacacac acatggcgct
     4141 taaagttgat ggtggaggtc atttgtcttg tagttttgtt accacttatc gttctaaaaa
     4201 gactgttggc aatatcaaaa tgccaggaat acatgctgta gaccacagac tagaaagact
     4261 cgaagagagc gataacgaaa tgttcgttgt acagagagag catgccgtag ccaaatttgc
     4321 tggcttaggc ggtggtatgg atgaattgta taagtaactc gagtggcctc ggtaccaaat
     4381 tccagaaaag acacccgaaa gggtgttttt tcgttttggt ccgctggaaa tctgctcgtc
     4441 agtgg
//
