LOCUS       sequence-323031-        4448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PJ23110-RBS-AbWS/DGAT-mRuby2-tL3S2P24_KanR_p15A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4448)
  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 4448)
  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: pLNL1130
FEATURES             Location/Qualifiers
     source          1..4448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(226..1041)
                     /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(226..1041)
                     /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     949..968
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1033..1052
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1319..1336)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1453..1998)
                     /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(1533..1552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2223..2257
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     CDS             3665..4372
                     /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 tctgctcgtc agtggtgctc acactgacga atcatgtaca gatcataccg atgactgcct
       61 ggcgactcac aactaagcaa gacagccgga accagcgccg gcgaacacca ctgcatatat
      121 ggcatatcac aacagtccac gtctcaagca gttacagaga tgttacgaac cactagtgca
      181 ctgcagtaca gcggccgcgt gttacaacca attaaccaat tctgattaga aaaactcatc
      241 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
      301 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
      361 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
      421 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
      481 tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
      541 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgaggcg
      601 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
      661 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
      721 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
      781 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
      841 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
      901 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
      961 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctgg agcaagacgt
     1021 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     1081 tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
     1141 aacgtggctt tgttgaataa atcgaacttt tgctgagttg aaggatcagt agcggagtgt
     1201 atactggctt actatgttgg cactgatgag ggtgtcagtg aagtgcttca tgtggcagga
     1261 gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata caggatatat tccgcttcct
     1321 cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac
     1381 ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc
     1441 ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc
     1501 aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg
     1561 ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg
     1621 gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc
     1681 tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc
     1741 gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt
     1801 gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt
     1861 tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga
     1921 accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc
     1981 agaccaaaac gatctcaaga agatcatctt attaatcaga taaaatattt gcggccgccc
     2041 ctgaattcgc atctagactg atgagacgtg gtagagccac aaacagccgg tacaagcaac
     2101 gatctccagg accatctgaa tcatgcgcgg atgacacgaa ctcacgacgg cgatcacaga
     2161 cattaaccca cagtacagac actgcgacaa cgtggcaatt cgtcgcaata caacggattt
     2221 actttacggc tagctcagtc ctaggtacaa tgctagcgct agcttcacac aggaaaccag
     2281 atctatgagg ccattgcatc caattgactt tatcttcttg tcattggaga aaagacaaca
     2341 accaatgcat gttggtggtt tgttcttatt tcagatacct gataacgctc cagatacatt
     2401 catccaagat ttagtcaacg atattaggat ctcaaaatct attccagtac caccattcaa
     2461 caataagtta aatggattgt tctgggatga agacgaagag tttgacttgg atcatcattt
     2521 tagacatatt gcattacctc atccaggaag aatcagggag ttgttgatct atatttcaca
     2581 ggaacactca acattattgg acagggctaa accattgtgg acatgtaaca taatcgaagg
     2641 tattgaggga aacagattcg ctatgtattt caaaatccac catgcaatgg tcgacggagt
     2701 cgcaggtatg agattgatag agaaatcttt atcacacgat gttacagaga aatcaatcgt
     2761 tcctccatgg tgtgtcgaag gtaagagagc taagagatta agagaaccaa agacaggaaa
     2821 gattaagaag attatgtcag gtatcaagtc tcagttgcaa gccactccaa ctgttattca
     2881 agaattgtct cagacagtat ttaaggatat cggaagaaat cctgaccatg tttcatcttt
     2941 ccaagctcct tgttctattt tgaatcaaag agtttcttca tcaagaaggt ttgctgctca
     3001 atctttcgat ttggataggt tcaggaatat cgccaaatct ttaaacgtca ctataaatga
     3061 tgttgttttg gcagtatgtt caggtgcttt gagggcttat ttgatgtcac ataattcatt
     3121 gccatcaaaa cctttgatcg ccatggttcc agcttcaatt agaaatgatg actcagacgt
     3181 ttctaacaga atcacaatga ttttagctaa tttggcaact cacaaagatg atccattaca
     3241 aagattggaa attattagaa ggtcagtcca aaattctaaa caaaggttta aaagaatgac
     3301 ttcagatcag attttgaatt actctgctgt cgtttacggt ccagctggat tgaatattat
     3361 ctcaggtatg atgcctaaga gacaggcctt caatttagtt atttcaaacg taccaggacc
     3421 tagagaacct ttatactgga atggtgcaaa attagatgct ttgtatcctg cttctatagt
     3481 cttggatgga caagctttga acatcactat gacttcttat ttggacaaat tagaagtagg
     3541 tttgatcgct tgcagaaatg ccttaccaag aatgcagaat ttattaactc atttagagga
     3601 agaaatacaa ttatttgagg gtgtcatagc caaacaagag gatattaaga ctgccaatgg
     3661 atccgtgtcc aaaggagagg agttaatcaa ggaaaacatg agaatgaaag ttgtcatgga
     3721 gggctccgtt aatggtcacc aattcaagtg tacaggggaa ggtgaaggta atccttacat
     3781 gggtacacaa actatgagaa ttaaagtaat tgaaggcgga ccactaccat ttgcatttga
     3841 cattctggca acgtcattca tgtacggatc acgaactttc atcaagtacc ctaaaggtat
     3901 accagacttt ttcaagcaat cttttccaga gggttttaca tgggaaaggg ttacaagata
     3961 cgaagatggg ggtgtcgtca cagttatgca agatacttca ttagaagatg gctgccttgt
     4021 ctatcatgtg caagtaagag gggtgaattt tccttctaac ggacctgtga tgcagaaaaa
     4081 gaccaaaggt tgggaaccaa atactgaaat gatgtaccca gctgatggag gtttgagagg
     4141 ctacacacac atggcgctta aagttgatgg tggaggtcat ttgtcttgta gttttgttac
     4201 cacttatcgt tctaaaaaga ctgttggcaa tatcaaaatg ccaggaatac atgctgtaga
     4261 ccacagacta gaaagactcg aagagagcga taacgaaatg ttcgttgtac agagagagca
     4321 tgccgtagcc aaatttgctg gcttaggcgg tggtatggat gaattgtata agtaactcga
     4381 gtggcctcgg taccaaattc cagaaaagac acccgaaagg gtgttttttc gttttggtcc
     4441 gctggaaa
//
