LOCUS       sequence-389808-        2646 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mKO2 under the control of PR promoter, p15A origin of 
            replication, Chloramphenicol selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2646)
  AUTHORS   Sarkar K, Bonnerjee D, Srivastava R, Bagh S
  TITLE     A single layer artificial neural network type architecture with 
            molecular engineered bacteria for reversible and irreversible 
            computing.
  JOURNAL   Chem Sci. 2021 Nov 9;12(48):15821-15832. doi: 10.1039/d1sc01505b. 
            eCollection 2021 Dec 15.
  PUBMED    35024106
REFERENCE   2  (bases 1 to 2646)
  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"; doi: 
            "10.1039/d1sc01505b"; journalName: "Chem Sci"; date: "2021-11-9- 9";
            volume: "12"; issue: "48"; pages: "15821-15832"
COMMENT     Sequence Label: PRC3MKO2
FEATURES             Location/Qualifiers
     source          1..2646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             71..82
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             89..745
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     terminator      757..843
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(878..895)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1012..1557)
                     /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(1092..1111)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1671..1765)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1786..2445)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2360..2379
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2446..2548)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     2614..2632
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagtaac accgtgcgtg ttgactattt tacctctggc ggtgataatg gttgcatgta
       61 cgaattcatt aaagaggaga aaggtaccat ggtgagtgtg attaaaccag agatgaagat
      121 gaggtactac atggacggct ccgtcaatgg gcatgagttc acaattgaag gtgaaggcac
      181 aggcagacct tacgagggac atcaagagat gacactacgc gtcacaatgg ccgagggcgg
      241 gccaatgcct ttcgcgtttg acttagtgtc acacgtgttc tgttacggcc acagagtatt
      301 tactaaatat ccagaagaga taccagacta tttcaaacaa gcatttcctg aaggcctgtc
      361 atgggaaagg tcgttggagt tcgaagatgg tgggtccgct tcagtcagtg cgcatataag
      421 ccttagagga aacaccttct accacaaatc caaatttact ggggttaact ttcctgccga
      481 tggtcctatc atgcaaaacc aaagtgttga ttgggagcca tcaaccgaga aaattactgc
      541 cagcgacgga gttctgaagg gtgatgttac gatgtaccta aaacttgaag gaggcggcaa
      601 tcacaaatgc caaatgaaga ctacttacaa ggcggcaaaa gagattcttg aaatgccagg
      661 agaccattac atcggccatc gcctcgtcag gaaaaccgaa ggcaacatta ctgagcaggt
      721 agaagatgca gtagctcata gctaatctag aggcatcaaa taaaacgaaa ggctcagtcg
      781 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
      841 aatccgccgc cctagaccta gggatatatt ccgcttcctc gctcactgac tcgctacgct
      901 cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat ttcctggaag
      961 atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag
     1021 gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc
     1081 gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt
     1141 tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac
     1201 gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc
     1261 ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa
     1321 gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga gttagtcttg
     1381 aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca
     1441 agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc
     1501 aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa
     1561 gatcatctta ttaatcagat aaaatatttc tagatttcag tgcaatttat ctcttcaaat
     1621 gtagcacctg aagtcagccc catacgatat aagttgttac tagtgcttgg attctcacca
     1681 ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc
     1741 attactggat ctatcaacag gagtccaagc gagctcgata tcaaattacg ccccgccctg
     1801 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
     1861 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
     1921 tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
     1981 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt
     2041 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
     2101 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
     2161 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     2221 catacggaat tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     2281 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
     2341 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca
     2401 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     2461 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     2521 aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccagatat cgacgtctaa
     2581 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     2641 cttcac
//
