LOCUS       sequence-260808-        4043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pZA4-oxySp-mCherry-TEVrs-LAA-pLsoxS-mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4043)
  AUTHORS   Muller IE, Rubens JR, Jun T, Graham D, Xavier R, Lu TK
  TITLE     Gene networks that compensate for crosstalk with crosstalk.
  JOURNAL   Nat Commun. 2019 Sep 6;10(1):4028. doi: 10.1038/s41467-019-12021-y.
  PUBMED    31492904
REFERENCE   2  (bases 1 to 4043)
  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: "Nat 
            Commun."; date: "2019-09-6"; volume: "10(1):4028. doi"; pages: " 
            10.1038/s41467-019-12021-y"
COMMENT     Sequence Label: pIM27
FEATURES             Location/Qualifiers
     source          1..4043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             111..122
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             129..836
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             843..863
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      941..1027
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             1125..1136
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1143..1853
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     terminator      1889..1975
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2012..2029)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2146..2691)
                     /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(2226..2245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2811..2905)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2955..3746)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 ttcattatcc atcctccatc gccacgatag ttcatggcga taggtagaat agcaatgaac
       61 gattatccct atcaagcatt ctgactgata attgctcaca cgaattcatt aaagaggaga
      121 aaggtaccat ggtgagcaag ggcgaagaag ataacatggc catcatcaag gagttcatgc
      181 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
      241 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
      301 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
      361 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
      421 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
      481 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
      541 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
      601 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
      661 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
      721 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
      781 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaaggtcg
      841 acgaaaacct gtacttccag ggtaggcctg cagcaaacga cgaaaactac gctttagcag
      901 cttaacccgg gggatcccat ggtacgcgtg ctagaggcat caaataaaac gaaaggctca
      961 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     1021 gacaaatccg ccgccctaga ctcgagccat aaatcgcttt acctcaagtt aacttgagga
     1081 attatactcc ccaacatcag caggacgcac tgaccgaatt cattaaagag gagaaaggta
     1141 ccatggtgag caagggcgaa gaagataaca tggccatcat caaggagttc atgcgcttca
     1201 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     1261 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     1321 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     1381 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     1441 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     1501 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     1561 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     1621 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     1681 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     1741 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     1801 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag taacccgggg
     1861 gatcccatgg tacgcgtgct agaggcatca aataaaacga aaggctcagt cgaaagactg
     1921 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     1981 gccctagacc tagggggata tattccgctt cctcgctcac tgactcgcta cgctcggtcg
     2041 ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg gaagatgcca
     2101 ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc ataggctccg
     2161 cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa acccgacagg
     2221 actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc ctgttcctgc
     2281 ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga
     2341 cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc
     2401 agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg
     2461 caaaagcacc actggcagca gccactggta attgatttag aggagttagt cttgaagtca
     2521 tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag
     2581 ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg
     2641 gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca agaagatcat
     2701 cttattaatc agataaaata tttctagatt tcagtgcaat ttatctcttc aaatgtagca
     2761 cctgaagtca gccccatacg atataagttg ttactagtac tagtgcttgg attctcacca
     2821 ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc
     2881 attactggat ctatcaacag gagtccaagc gagctcgagc tcgatatccg tcggcttgaa
     2941 cgaattgtta gacattattt gccgactacc ttggtgatct cgcctttcac gtagtggaca
     3001 aattcttcca actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc
     3061 tgtctagctt caagtatgac gggctgatac tgggccggca ggcgctccat tgcccagtcg
     3121 gcagcgacat ccttcggcgc gattttgccg gttactgcgc tgtaccaaat gcgggacaac
     3181 gtaagcacta catttcgctc atcgccagcc cagtcgggcg gcgagttcca tagcgttaag
     3241 gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc
     3301 gctggaccta ccaaggcaac gctatgttct cttgcttttg tcagcaagat agccagatca
     3361 atgtcgatcg tggctggctc gaagatacct gcaagaatgt cattgcgctg ccattctcca
     3421 aattgcagtt cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg cacaacaatg
     3481 gtgacttcta cagcgcggag aatctcgctc tctccagggg aagccgaagt ttccaaaagg
     3541 tcgttgatca aagctcgccg cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa
     3601 tcaatatcac tgtgtggctt caggccgcca tccactgcgg agccgtacaa atgtacggcc
     3661 agcaacgtcg gttcgagatg gcgctcgatg acgccaacta cctctgatag ttgagtcgat
     3721 acttcggcga tcaccgcttc cctcatgatg tttaactttg ttttagggcg actgccctgc
     3781 tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc
     3841 tgcttggatg cccgaggcat agactgtacc ccaaaaaaac agtcataaca agccatgaaa
     3901 accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag
     3961 cgcatacgct acttgcatta cagcttacga accgaacagg cttatgtcca ctgggttcgt
     4021 gccttcatcc ggatatcgac gtc
//
