LOCUS       sequence-323029-        4247 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PJ23110-RBS-FaAAT2-mRuby2-tL3S2P24_KanR_p15A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4247)
  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 4247)
  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: pLNL1126
FEATURES             Location/Qualifiers
     source          1..4247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(227..1042)
                     /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(227..1042)
                     /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     950..969
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1034..1053
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1320..1337)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1454..1999)
                     /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(1534..1553)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2224..2258
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     CDS             3465..4172
                     /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 atctgctcgt cagtggtgct cacactgacg aatcatgtac agatcatacc gatgactgcc
       61 tggcgactca caactaagca agacagccgg aaccagcgcc ggcgaacacc actgcatata
      121 tggcatatca caacagtcca cgtctcaagc agttacagag atgttacgaa ccactagtgc
      181 actgcagtac agcggccgcg tgttacaacc aattaaccaa ttctgattag aaaaactcat
      241 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
      301 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
      361 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
      421 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
      481 atggcaaaag cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
      541 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgaggc
      601 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
      661 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
      721 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
      781 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
      841 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
      901 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
      961 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctg gagcaagacg
     1021 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     1081 ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca
     1141 caacgtggct ttgttgaata aatcgaactt ttgctgagtt gaaggatcag tagcggagtg
     1201 tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc atgtggcagg
     1261 agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata ttccgcttcc
     1321 tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa
     1381 cggggcggag atttcctgga agatgccagg aagatactta acagggaagt gagagggccg
     1441 cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct
     1501 caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg
     1561 gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat
     1621 ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag
     1681 ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat
     1741 cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat
     1801 tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag
     1861 ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag
     1921 aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg
     1981 cagaccaaaa cgatctcaag aagatcatct tattaatcag ataaaatatt tgcggccgcc
     2041 cctgaattcg catctagact gatgagacgt ggtagagcca caaacagccg gtacaagcaa
     2101 cgatctccag gaccatctga atcatgcgcg gatgacacga actcacgacg gcgatcacag
     2161 acattaaccc acagtacaga cactgcgaca acgtggcaat tcgtcgcaat acaacggatt
     2221 tactttacgg ctagctcagt cctaggtaca atgctagcgc tagcttcaca caggaaacca
     2281 gatctatgtc ttataagaat aaccattcta ttttatcaaa accaaatgat ccagtcgaag
     2341 taattagaga tgcattatct aaggctttac agttctacta cccattggcc ggtagattga
     2401 gagaaggtcc aaataagaag ttgatggttg actgtaccgg agaaggtatt ttatttgtag
     2461 aggctaatgc cgaagttacc ttagatgagt taggtgatgc tatcttacca ccatgcccat
     2521 tcttagatgg tttcttattt aacgttccag gttcagatgg tattttaggt tctccattgt
     2581 gtttgataca agtaactaga ttgtcatgtg gtggattcat attcgcatta agattgaacc
     2641 atactatctg cgatgctttg ggtttagttc agttcttgaa cgctgtaggt gaaatagctc
     2701 aaggtaagta cgcaccatct ataacccctg tttgggaaag agagttatta tcagcaagag
     2761 atcctccaag aatctcatgc acacatgagg aatttgacga ttctattgat cactcttacc
     2821 caaattatgg agctactgtt cagcagtgtt actgttttgg tccaaaggag atcaaatctt
     2881 tgagagaaca tttgccacca catttgtcta catgttcttc tacttttgaa ttgatcaccg
     2941 cctgcgtttg gaagtgcaga actatatcat tggatatgga cccagaacag atagtaagat
     3001 tgtcttgtgt agtaaccgcc ttaggtaagc ataacaacgt ttgcttgcca ttgggttatt
     3061 acggtaatac tttcacctat ccagcagtcg tttctaccgc cgagaggttg tgtaactcac
     3121 ctttaggtta cgcagtagaa ttggtcaaga aatctaaggc taaaatgtct gaagagtatt
     3181 taagatcagc catagatttt gttgaggtca ggggaagacc tccattcgcc ttagaaggta
     3241 tgtcagattt cttagtttct gataatacaa gaactggttt gggtgaaata gactttggtt
     3301 ttggtaaacc agtttacgct ggtgttgcta aatcaaccga tttaatctca ttttatgtaa
     3361 ggtcaacaaa taaggaggaa agagaaatct tagtccctgt ttgtttgcca atattgtcaa
     3421 tggagatatt tcaacaagag ttaaagaaga tgatcggagg atccgtgtcc aaaggagagg
     3481 agttaatcaa ggaaaacatg agaatgaaag ttgtcatgga gggctccgtt aatggtcacc
     3541 aattcaagtg tacaggggaa ggtgaaggta atccttacat gggtacacaa actatgagaa
     3601 ttaaagtaat tgaaggcgga ccactaccat ttgcatttga cattctggca acgtcattca
     3661 tgtacggatc acgaactttc atcaagtacc ctaaaggtat accagacttt ttcaagcaat
     3721 cttttccaga gggttttaca tgggaaaggg ttacaagata cgaagatggg ggtgtcgtca
     3781 cagttatgca agatacttca ttagaagatg gctgccttgt ctatcatgtg caagtaagag
     3841 gggtgaattt tccttctaac ggacctgtga tgcagaaaaa gaccaaaggt tgggaaccaa
     3901 atactgaaat gatgtaccca gctgatggag gtttgagagg ctacacacac atggcgctta
     3961 aagttgatgg tggaggtcat ttgtcttgta gttttgttac cacttatcgt tctaaaaaga
     4021 ctgttggcaa tatcaaaatg ccaggaatac atgctgtaga ccacagacta gaaagactcg
     4081 aagagagcga taacgaaatg ttcgttgtac agagagagca tgccgtagcc aaatttgctg
     4141 gcttaggcgg tggtatggat gaattgtata agtaactcga gtggcctcgg taccaaattc
     4201 cagaaaagac acccgaaagg gtgttttttc gttttggtcc gctggaa
//
