LOCUS       sequence-389810-        2667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses E2-Crimson under the control of PLUX* promoter, p15A 
            origin of replication, Chloramphenicol selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2667)
  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 2667)
  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: pX*C3E2-Crimson
FEATURES             Location/Qualifiers
     source          1..2667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             71..82
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             89..766
                     /codon_start=1
                     /product="far-red noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2009)"
                     /label=E2-Crimson
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGVGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFFYGSKAYIKHPADIPDYLKQSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTLIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERNYPRDGVLKGENHM
                     ALKLKGGGHYLCEFKSIYMAKKPVKLPGYHYVDYKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     487..506
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      778..864
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(899..916)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1033..1578)
                     /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(1113..1132)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1692..1786)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1807..2466)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2381..2400
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2467..2569)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     2635..2653
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagacct gtaggatcgt acaggtttac gcaagaaaat ggtttgttac tttcgaataa
       61 agaattcatt aaagaggaga aaggtaccat ggatagcact gagaacgtca tcaagccctt
      121 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
      181 cgtgggcgag ggcaagccct acgagggcac ccagaccgcc aagctgcaag tgaccaaggg
      241 cggccccctg cccttcgcct gggacatcct gtccccccag ttcttctacg gctccaaggc
      301 gtacatcaag caccccgccg acatccccga ctacctcaag cagtccttcc ccgagggctt
      361 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
      421 ctccctgcag gacggcaccc tcatctacca cgtgaagttc atcggcgtga acttcccctc
      481 cgacggcccc gtaatgcaga agaagactct gggctgggag ccctccactg agcgcaacta
      541 cccccgcgac ggcgtgctga agggcgagaa ccacatggcg ctgaagctga agggcggcgg
      601 ccactacctg tgtgagttca agtccatcta catggccaag aagcccgtga agctgcccgg
      661 ctaccactac gtggactaca agctcgacat cacctcccac aacgaggact acaccgtggt
      721 ggagcagtac gagcgcgccg aggcccgcca ccacctgttc cagtagtcta gaggcatcaa
      781 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
      841 aacgctctcc tgagtaggac aaatccgccg ccctagacct agggatatat tccgcttcct
      901 cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac
      961 ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc
     1021 ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc
     1081 aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg
     1141 ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg
     1201 gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc
     1261 tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc
     1321 gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt
     1381 gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt
     1441 tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga
     1501 accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc
     1561 agaccaaaac gatctcaaga agatcatctt attaatcaga taaaatattt ctagatttca
     1621 gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtta
     1681 ctagtgcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa
     1741 tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctcgat
     1801 atcaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct
     1861 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
     1921 cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat
     1981 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
     2041 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
     2101 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
     2161 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
     2221 cagctcaccg tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag
     2281 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     2341 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
     2401 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     2461 ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     2521 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa cgtctcattt
     2581 tcgccagata tcgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     2641 cgtatcacga ggccctttcg tcttcac
//
