LOCUS       sequence-323269-        3737 bp    DNA     circular SYN 15-APR-2025
DEFINITION  PJ23110-RBS-CAT-taa-mRuby2-tL3S2P24_KanR_p15A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3737)
  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 3737)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pLNL1135
FEATURES             Location/Qualifiers
     source          1..3737
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(246..1061)
                     /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(246..1061)
                     /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     969..988
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1053..1072
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1339..1356)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1473..2018)
                     /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(1553..1572)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2243..2277
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     CDS             2308..2967
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2374..2393)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2974..3681
                     /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 tcgttttggt ccgctggaaa tctgctcgtc agtggtgctc acactgacga atcatgtaca
       61 gatcataccg atgactgcct ggcgactcac aactaagcaa gacagccgga accagcgccg
      121 gcgaacacca ctgcatatat ggcatatcac aacagtccac gtctcaagca gttacagaga
      181 tgttacgaac cactagtgca ctgcagtaca gcggccgcgt gttacaacca attaaccaat
      241 tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
      301 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
      361 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
      421 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
      481 acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca
      541 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
      601 gattgcgcct gagcgaggcg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
      661 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
      721 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
      781 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
      841 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
      901 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
      961 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     1021 cgcggcctgg agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
     1081 tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa
     1141 catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg
     1201 aaggatcagt agcggagtgt atactggctt actatgttgg cactgatgag ggtgtcagtg
     1261 aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata
     1321 caggatatat tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg
     1381 agcggaaatg gcttacgaac ggggcggaga tttcctggaa gatgccagga agatacttaa
     1441 cagggaagtg agagggccgc ggcaaagccg tttttccata ggctccgccc ccctgacaag
     1501 catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc cgacaggact ataaagatac
     1561 caggcgtttc cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc
     1621 ggtgtcattc cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagttccgg
     1681 gtaggcagtt cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg
     1741 cgccttatcc ggtaactatc gtcttgagtc caacccggaa agacatgcaa aagcaccact
     1801 ggcagcagcc actggtaatt gatttagagg agttagtctt gaagtcatgc gccggttaag
     1861 gctaaactga aaggacaagt tttggtgact gcgctcctcc aagccagtta cctcggttca
     1921 aagagttggt agctcagaga accttcgaaa aaccgccctg caaggcggtt ttttcgtttt
     1981 cagagcaaga gattacgcgc agaccaaaac gatctcaaga agatcatctt attaatcaga
     2041 taaaatattt gcggccgccc ctgaattcgc atctagactg atgagacgtg gtagagccac
     2101 aaacagccgg tacaagcaac gatctccagg accatctgaa tcatgcgcgg atgacacgaa
     2161 ctcacgacgg cgatcacaga cattaaccca cagtacagac actgcgacaa cgtggcaatt
     2221 cgtcgcaata caacggattt actttacggc tagctcagtc ctaggtacaa tgctagcgct
     2281 agcttcacac aggaaaccag atctatgatg gagaaaaaaa tcactggata taccaccgtt
     2341 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
     2401 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
     2461 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg
     2521 gaatttcgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
     2581 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
     2641 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
     2701 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tttcagccaa tccctgggtg
     2761 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
     2821 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
     2881 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
     2941 tgcgatgagt ggcagggcgg ggcgtaagga tccgtgtcca aaggagagga gttaatcaag
     3001 gaaaacatga gaatgaaagt tgtcatggag ggctccgtta atggtcacca attcaagtgt
     3061 acaggggaag gtgaaggtaa tccttacatg ggtacacaaa ctatgagaat taaagtaatt
     3121 gaaggcggac cactaccatt tgcatttgac attctggcaa cgtcattcat gtacggatca
     3181 cgaactttca tcaagtaccc taaaggtata ccagactttt tcaagcaatc ttttccagag
     3241 ggttttacat gggaaagggt tacaagatac gaagatgggg gtgtcgtcac agttatgcaa
     3301 gatacttcat tagaagatgg ctgccttgtc tatcatgtgc aagtaagagg ggtgaatttt
     3361 ccttctaacg gacctgtgat gcagaaaaag accaaaggtt gggaaccaaa tactgaaatg
     3421 atgtacccag ctgatggagg tttgagaggc tacacacaca tggcgcttaa agttgatggt
     3481 ggaggtcatt tgtcttgtag ttttgttacc acttatcgtt ctaaaaagac tgttggcaat
     3541 atcaaaatgc caggaataca tgctgtagac cacagactag aaagactcga agagagcgat
     3601 aacgaaatgt tcgttgtaca gagagagcat gccgtagcca aatttgctgg cttaggcggt
     3661 ggtatggatg aattgtataa gtaactcgag tggcctcggt accaaattcc agaaaagaca
     3721 cccgaaaggg tgttttt
//
