LOCUS       sequence-323259-        4427 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PJ23110-RBS-ScEHT1-mRuby2-tL3S2P24_KanR_p15A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4427)
  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 4427)
  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: pLNL1134
FEATURES             Location/Qualifiers
     source          1..4427
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(194..1009)
                     /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(194..1009)
                     /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     917..936
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1001..1020
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1287..1304)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1421..1966)
                     /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(1501..1520)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2191..2225
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     CDS             3612..4319
                     /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 catgtacaga tcataccgat gactgcctgg cgactcacaa ctaagcaaga cagccggaac
       61 cagcgccggc gaacaccact gcatatatgg catatcacaa cagtccacgt ctcaagcagt
      121 tacagagatg ttacgaacca ctagtgcact gcagtacagc ggccgcgtgt tacaaccaat
      181 taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
      241 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
      301 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
      361 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
      421 catgagtgac gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt
      481 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
      541 tcattcgtga ttgcgcctga gcgaggcgaa atacgcgatc gctgttaaaa ggacaattac
      601 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
      661 ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga
      721 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
      781 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
      841 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
      901 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
      961 aatttaatcg cggcctggag caagacgttt cccgttgaat atggctcata acaccccttg
     1021 tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg
     1081 caatgtaaca tcagagattt tgagacacaa cgtggctttg ttgaataaat cgaacttttg
     1141 ctgagttgaa ggatcagtag cggagtgtat actggcttac tatgttggca ctgatgaggg
     1201 tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg tcagcagaat
     1261 atgtgataca ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga
     1321 ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag
     1381 atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc
     1441 ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat
     1501 aaagatacca ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc
     1561 ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc
     1621 agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc
     1681 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa
     1741 gcaccactgg cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc
     1801 cggttaaggc taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc
     1861 tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt
     1921 ttcgttttca gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat
     1981 taatcagata aaatatttgc ggccgcccct gaattcgcat ctagactgat gagacgtggt
     2041 agagccacaa acagccggta caagcaacga tctccaggac catctgaatc atgcgcggat
     2101 gacacgaact cacgacggcg atcacagaca ttaacccaca gtacagacac tgcgacaacg
     2161 tggcaattcg tcgcaataca acggatttac tttacggcta gctcagtcct aggtacaatg
     2221 ctagcgctag cttcacacag gaaaccagat ctatgtcaga agtttccaaa tggccagcaa
     2281 tcaacccatt ccattgggga tacaatggta cagtttcgca tattgtcggt gaaaatggtt
     2341 ccattaaact ccatttaaaa gacaacaagg agcaagttga ttttgacgag ttcgctaaca
     2401 aatatgtccc aacgttgaag aatggtgccc aattcaaatt gagtccttac ttgttcacag
     2461 gtattttgca aactttgtac ttaggtgctg ctgatttctc taagaaattt cctgtattct
     2521 acggcaggga aattgtcaaa ttctcggatg gtggagtttg caccgctgac tggctcatag
     2581 attcatggaa aaaggattat gaattcgatc aaagtactac gagctttgat aaaaaaaaat
     2641 ttgataaaga cgagaaggcg acacatccag aaggatggcc tcgtttacaa ccacgtacaa
     2701 ggtacctgaa agataatgag ttggaagaac tacgggaggt tgatctaccc ctagtagtta
     2761 ttctacatgg tcttgctggt ggtagtcatg agccgattat aagatctctt gctgaaaacc
     2821 tgtctcgcag tgggagattt caagtggtcg tcctaaatac cagaggttgt gcacgttcca
     2881 aaattaccac cagaaattta tttacagctt atcacacaat ggatattcgc gagtttttgc
     2941 aaagagaaaa gcaaagacat ccagatagaa aactatacgc tgtgggatgc tcttttggtg
     3001 ctacgatgct ggcaaactat ctgggagaag agggcgataa atcaccttta tccgcagctg
     3061 ctactttgtg caatccttgg gatcttctcc tttcagcaat taggatgagc caggattggt
     3121 ggtcaagaac tttattttcc aaaaatattg cgcaattctt aacaagaacc gttcaggtta
     3181 atatgggtga attaggagtt ccaaatggct ctctccccga tcatcctccc acagtcaaga
     3241 atccatcttt ctatatgttc acgcctgaaa atctaataaa ggcaaagagc tttaaatcga
     3301 cccgggaatt tgatgaagtg tacactgcgc ctgctttagg cttcccaaat gctatggagt
     3361 attataaagc ggccagctca ataaacagag ttgatacaat tcgggttcct acccttgtta
     3421 tcaattccag ggatgatcct gttgtcggcc cagatcaacc atactcaatc gtggaaaaga
     3481 atcctcgtat tttgtattgt agaaccgatt taggtggtca tttagcttac ctagataaag
     3541 acaacaactc gtgggctacc aaggcaattg cagaattttt cactaagttt gatgaattag
     3601 tcgtaggatc cgtgtccaaa ggagaggagt taatcaagga aaacatgaga atgaaagttg
     3661 tcatggaggg ctccgttaat ggtcaccaat tcaagtgtac aggggaaggt gaaggtaatc
     3721 cttacatggg tacacaaact atgagaatta aagtaattga aggcggacca ctaccatttg
     3781 catttgacat tctggcaacg tcattcatgt acggatcacg aactttcatc aagtacccta
     3841 aaggtatacc agactttttc aagcaatctt ttccagaggg ttttacatgg gaaagggtta
     3901 caagatacga agatgggggt gtcgtcacag ttatgcaaga tacttcatta gaagatggct
     3961 gccttgtcta tcatgtgcaa gtaagagggg tgaattttcc ttctaacgga cctgtgatgc
     4021 agaaaaagac caaaggttgg gaaccaaata ctgaaatgat gtacccagct gatggaggtt
     4081 tgagaggcta cacacacatg gcgcttaaag ttgatggtgg aggtcatttg tcttgtagtt
     4141 ttgttaccac ttatcgttct aaaaagactg ttggcaatat caaaatgcca ggaatacatg
     4201 ctgtagacca cagactagaa agactcgaag agagcgataa cgaaatgttc gttgtacaga
     4261 gagagcatgc cgtagccaaa tttgctggct taggcggtgg tatggatgaa ttgtataagt
     4321 aactcgagtg gcctcggtac caaattccag aaaagacacc cgaaagggtg ttttttcgtt
     4381 ttggtccgct ggaaatctgc tcgtcagtgg tgctcacact gacgaat
//
