LOCUS       sequence-323035-        4442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PJ23110-RBS-ScEEB1-mRuby2-tL3S2P24_KanR_p15A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4442)
  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 4442)
  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: pLNL1133
FEATURES             Location/Qualifiers
     source          1..4442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(219..1034)
                     /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(219..1034)
                     /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     942..961
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1026..1045
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1312..1329)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1446..1991)
                     /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(1526..1545)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2216..2250
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     CDS             3652..4359
                     /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 gtcagtggtg ctcacactga cgaatcatgt acagatcata ccgatgactg cctggcgact
       61 cacaactaag caagacagcc ggaaccagcg ccggcgaaca ccactgcata tatggcatat
      121 cacaacagtc cacgtctcaa gcagttacag agatgttacg aaccactagt gcactgcagt
      181 acagcggccg cgtgttacaa ccaattaacc aattctgatt agaaaaactc atcgagcatc
      241 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
      301 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
      361 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
      421 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
      481 agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
      541 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag gcgaaatacg
      601 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
      661 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
      721 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
      781 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
      841 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
      901 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
      961 ccatataaat cagcatccat gttggaattt aatcgcggcc tggagcaaga cgtttcccgt
     1021 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt
     1081 catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg
     1141 ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc agtagcggag tgtatactgg
     1201 cttactatgt tggcactgat gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa
     1261 ggctgcaccg gtgcgtcagc agaatatgtg atacaggata tattccgctt cctcgctcac
     1321 tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg aacggggcgg
     1381 agatttcctg gaagatgcca ggaagatact taacagggaa gtgagagggc cgcggcaaag
     1441 ccgtttttcc ataggctccg cccccctgac aagcatcacg aaatctgacg ctcaaatcag
     1501 tggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg cggctccctc
     1561 gtgcgctctc ctgttcctgc ctttcggttt accggtgtca ttccgctgtt atggccgcgt
     1621 ttgtctcatt ccacgcctga cactcagttc cgggtaggca gttcgctcca agctggactg
     1681 tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta tccggtaact atcgtcttga
     1741 gtccaacccg gaaagacatg caaaagcacc actggcagca gccactggta attgatttag
     1801 aggagttagt cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca agttttggtg
     1861 actgcgctcc tccaagccag ttacctcggt tcaaagagtt ggtagctcag agaaccttcg
     1921 aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca agagattacg cgcagaccaa
     1981 aacgatctca agaagatcat cttattaatc agataaaata tttgcggccg cccctgaatt
     2041 cgcatctaga ctgatgagac gtggtagagc cacaaacagc cggtacaagc aacgatctcc
     2101 aggaccatct gaatcatgcg cggatgacac gaactcacga cggcgatcac agacattaac
     2161 ccacagtaca gacactgcga caacgtggca attcgtcgca atacaacgga tttactttac
     2221 ggctagctca gtcctaggta caatgctagc gctagcttca cacaggaaac cagatctatg
     2281 tttcgctcgg gttactatcc aactgtcact cctagccatt ggggctataa cggcaccgtc
     2341 aaacatgtac tgggagagaa aggaaccaag tctttggctt tcagagattc taagcgccaa
     2401 attcctcttc atgaattcgt aaccaaacac gtgcctacac tcaaagatgg tgcaaatttt
     2461 aggttaaaca gcttgctttt cacaggttac ttacagaccc tttatttgtc agccggtgat
     2521 ttttccaaaa aatttcaggt attttatgga agagagatta taaaattttc tgatggaggg
     2581 gtctgcactg cagattgggt tatgcctgaa tgggagcaaa catattcatt aaatgctgaa
     2641 aaggctagtt ttaacgagaa acaattttcg aacgatgaaa aggccaccca tccaaagggt
     2701 tggcctcgtc ttcacccgag aactagatat ctatcctctg aggaacttga aaaatgtcat
     2761 tctaagggtt attcttaccc tttagtggta gtacttcatg gtcttgcagg gggaagccat
     2821 gagcctctca tcagggcatt atctgaagat ctatcaaaag tcggtgacgg taaattccaa
     2881 gtggtagtcc ttaatgctag aggttgctca agatctaaag tcaccacccg tagaatcttt
     2941 accgcattac acacaggtga tgtgagagaa tttttgaacc accaaaaagc tctatttccg
     3001 caaagaaaaa tatatgcagt cggaacatca tttggagcag ccatgctaac gaactactta
     3061 ggtgaagaag gagataactg tccattaaat gctgcagtcg ctctctcaaa cccatgggat
     3121 ttcgtacaca cttgggacaa gttggctcat gattggtggt ccaaccatat tttttctaga
     3181 actttaacac aatttctaac aaggactgtc aaggttaata tgaatgagtt gcaggtacca
     3241 gagaattttg aggtttcgca caaaccgaca gttgagaaac cagtctttta tacgtatacc
     3301 agagaaaatt tggaaaaggc tgaaaaattt acagacatat tagaattcga taatcttttc
     3361 actgccccat cgatgggcct gccggacggg ttaacatatt atagaaaggc cagttcgata
     3421 aatagattgc ctaatatcaa aatcccaact ttaataatca acgcaacgga tgatccagtt
     3481 acaggtgaaa acgtaattcc gtacaaacaa gctagagaaa atccttgtgt gttgttatgc
     3541 gagactgact tgggtggaca tttggcttat ctcgacaatg aaagtaattc ctggttaaca
     3601 aagcaagctg ccgagttctt gggcagcttt gatgagttag ttttaggatc cgtgtccaaa
     3661 ggagaggagt taatcaagga aaacatgaga atgaaagttg tcatggaggg ctccgttaat
     3721 ggtcaccaat tcaagtgtac aggggaaggt gaaggtaatc cttacatggg tacacaaact
     3781 atgagaatta aagtaattga aggcggacca ctaccatttg catttgacat tctggcaacg
     3841 tcattcatgt acggatcacg aactttcatc aagtacccta aaggtatacc agactttttc
     3901 aagcaatctt ttccagaggg ttttacatgg gaaagggtta caagatacga agatgggggt
     3961 gtcgtcacag ttatgcaaga tacttcatta gaagatggct gccttgtcta tcatgtgcaa
     4021 gtaagagggg tgaattttcc ttctaacgga cctgtgatgc agaaaaagac caaaggttgg
     4081 gaaccaaata ctgaaatgat gtacccagct gatggaggtt tgagaggcta cacacacatg
     4141 gcgcttaaag ttgatggtgg aggtcatttg tcttgtagtt ttgttaccac ttatcgttct
     4201 aaaaagactg ttggcaatat caaaatgcca ggaatacatg ctgtagacca cagactagaa
     4261 agactcgaag agagcgataa cgaaatgttc gttgtacaga gagagcatgc cgtagccaaa
     4321 tttgctggct taggcggtgg tatggatgaa ttgtataagt aactcgagtg gcctcggtac
     4381 caaattccag aaaagacacc cgaaagggtg ttttttcgtt ttggtccgct ggaaatctgc
     4441 tc
//
