LOCUS       sequence-323300-        4430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PJ23110-RBS-FvSAAT-mRuby2-tL3S2P24_KanR_p15A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4430)
  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 4430)
  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: pLNL1142
FEATURES             Location/Qualifiers
     source          1..4430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(259..1074)
                     /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(259..1074)
                     /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     982..1001
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1066..1085
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1352..1369)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1486..2031)
                     /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(1566..1585)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2256..2290
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     CDS             3680..4387
                     /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 aaagggtgtt ttttcgtttt ggtccgctgg aaatctgctc gtcagtggtg ctcacactga
       61 cgaatcatgt acagatcata ccgatgactg cctggcgact cacaactaag caagacagcc
      121 ggaaccagcg ccggcgaaca ccactgcata tatggcatat cacaacagtc cacgtctcaa
      181 gcagttacag agatgttacg aaccactagt gcactgcagt acagcggccg cgtgttacaa
      241 ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
      301 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
      361 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
      421 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
      481 atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca
      541 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
      601 gttattcatt cgtgattgcg cctgagcgag gcgaaatacg cgatcgctgt taaaaggaca
      661 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
      721 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
      781 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
      841 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
      901 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
      961 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     1021 gttggaattt aatcgcggcc tggagcaaga cgtttcccgt tgaatatggc tcataacacc
     1081 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
     1141 ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac
     1201 ttttgctgag ttgaaggatc agtagcggag tgtatactgg cttactatgt tggcactgat
     1261 gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc
     1321 agaatatgtg atacaggata tattccgctt cctcgctcac tgactcgcta cgctcggtcg
     1381 ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg gaagatgcca
     1441 ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc ataggctccg
     1501 cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa acccgacagg
     1561 actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc ctgttcctgc
     1621 ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga
     1681 cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc
     1741 agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg
     1801 caaaagcacc actggcagca gccactggta attgatttag aggagttagt cttgaagtca
     1861 tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag
     1921 ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg
     1981 gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca agaagatcat
     2041 cttattaatc agataaaata tttgcggccg cccctgaatt cgcatctaga ctgatgagac
     2101 gtggtagagc cacaaacagc cggtacaagc aacgatctcc aggaccatct gaatcatgcg
     2161 cggatgacac gaactcacga cggcgatcac agacattaac ccacagtaca gacactgcga
     2221 caacgtggca attcgtcgca atacaacgga tttactttac ggctagctca gtcctaggta
     2281 caatgctagc gctagcttca cacaggaaac cagatctatg gagaaaatcg aggtttcgat
     2341 caattcgaaa catacgatta aacctagtac ctcgtccacc ccgctgcagc cgtataaact
     2401 gactctgctg gatcaactga ccccgcctgc atacgtaccg atcgtgttct tctacccgat
     2461 cacagatcat gattttaacc ttcctcaaac tctggccgat ctgcgtcagg cactgagcga
     2521 aaccctgacc ctttactatc cgctgagtgg ccgcgtgaag aacaacctgt atatcgatga
     2581 ttttgaagaa ggcgttccgt atcttgaagc tcgcgtgaac tgcgatatga ccgattttct
     2641 tcgcctgcgc aagattgaat gcctgaacga attcgttccc atcaaaccat tctccatgga
     2701 ggcgatttcc gatgaacgtt acccgctgct gggagtacaa gtcaacgtat ttgattcagg
     2761 tattgctatt ggtgttagcg tcagccataa attgatcgat gggggtaccg cggattgctt
     2821 tctgaagagc tggggagcgg tgttccgcgg gtgccgcgag aacatcattc accccagtct
     2881 gagtgaagca gcgctgctgt tcccgccgcg cgacgatctg cctgaaaaat acgtcgatca
     2941 aatggaagct ctctggttcg cggggaagaa agtggcgacg cgccgtttcg ttttcggtgt
     3001 caaggccatt tctagcattc aggacgaagc caagtcagaa tcagtcccca aaccatctcg
     3061 cgtgcatgca gtcaccggct tcctgtggaa gcacctgatc gcggccagcc gcgcgttgac
     3121 gtccggcacc accagcaccc gcctctccat cgcagctcag gcagttaacc tccgcacccg
     3181 catgaacatg gaaacggtac tcgataacgc gacaggtaac ttattttggt gggcacaggc
     3241 aattctggaa cttagccata ccacgccgga aatcagcgat cttaaactgt gcgatttggt
     3301 taatcttctg aatggcagcg tgaagcagtg caatggtgat tactttgaaa ccttcaaagg
     3361 taaggagggt tacggccgca tgtgcgaata cctggatttc caacgcacga tgtcttccat
     3421 ggaaccggcg cctgatattt atctgttttc tagttggacc aatttcttca acccgttaga
     3481 ttttggttgg ggccgcacca gttggatcgg tgtggccgga aagattgaat ctgcctcttg
     3541 taagttcatt attctggtac caacccagtg tggctctggg atcgaagcgt gggtgaacct
     3601 cgaagaagaa aaaatggcaa tgttggaaca ggatccgcat ttccttgcgt tagccagccc
     3661 gaaaacgctg atcggatccg tgtccaaagg agaggagtta atcaaggaaa acatgagaat
     3721 gaaagttgtc atggagggct ccgttaatgg tcaccaattc aagtgtacag gggaaggtga
     3781 aggtaatcct tacatgggta cacaaactat gagaattaaa gtaattgaag gcggaccact
     3841 accatttgca tttgacattc tggcaacgtc attcatgtac ggatcacgaa ctttcatcaa
     3901 gtaccctaaa ggtataccag actttttcaa gcaatctttt ccagagggtt ttacatggga
     3961 aagggttaca agatacgaag atgggggtgt cgtcacagtt atgcaagata cttcattaga
     4021 agatggctgc cttgtctatc atgtgcaagt aagaggggtg aattttcctt ctaacggacc
     4081 tgtgatgcag aaaaagacca aaggttggga accaaatact gaaatgatgt acccagctga
     4141 tggaggtttg agaggctaca cacacatggc gcttaaagtt gatggtggag gtcatttgtc
     4201 ttgtagtttt gttaccactt atcgttctaa aaagactgtt ggcaatatca aaatgccagg
     4261 aatacatgct gtagaccaca gactagaaag actcgaagag agcgataacg aaatgttcgt
     4321 tgtacagaga gagcatgccg tagccaaatt tgctggctta ggcggtggta tggatgaatt
     4381 gtataagtaa ctcgagtggc ctcggtacca aattccagaa aagacacccg
//
