LOCUS       sequence-420216-        2731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses E2crimson.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2731)
  AUTHORS   Srivastava R, Sarkar K, Bonnerjee D, Bagh S
  TITLE     Synthetic Genetic Reversible Feynman Gate in a Single E. coli Cell 
            and Its Application in Bacterial to Mammalian Cell Information 
            Transfer.
  JOURNAL   ACS Synth Biol. 2022 Mar 18;11(3):1040-1048. doi: 
            10.1021/acssynbio.1c00392. Epub 2022 Feb 18.
  PUBMED    35179369
REFERENCE   2  (bases 1 to 2731)
  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: 
            "10.1021/acssynbio.1c00392"; journalName: "ACS Synth Biol"; date: 
            "2022-03-18- 18"; volume: "11"; issue: "3"; pages: "1040-1048"
COMMENT     Sequence Label: pNB_AR_Pllaco 1E2crimson
FEATURES             Location/Qualifiers
     source          1..2731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      178..272
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      386..931
                     /direction=RIGHT
                     /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     832..851
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1142..1158
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    1165..1181
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             1223..1234
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1241..1918
                     /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     1639..1658
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      1930..2016
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        2126..2228
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(2229..2731,1..157)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2295..2314)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
       61 gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
      121 acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt gatatcgagc tcgcttggac
      181 tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg
      241 gttgccgccg ggcgtttttt attggtgaga atccaagcac tagtaacaac ttatatcgta
      301 tggggctgac ttcaggtgct acatttgaag agataaattg cactgaaatc tagaaatatt
      361 ttatctgatt aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct
      421 ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga gctaccaact
      481 ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc ctttcagttt
      541 agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc agtggctgct
      601 gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt accggataag
      661 gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga gcgaactgcc
      721 tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca gcggaatgac
      781 accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag ggggaaacgc
      841 ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc agatttcgtg
      901 atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc tctcacttcc
      961 ctgttaagta tcttcctggc atcttccaga catgttcttt cctgcgttat cccctgattc
     1021 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacggt
     1081 tgttttggag cacggaactg cagaatgaat agcttctgcc aggatcctgg taactcgaga
     1141 attgtgagcg gataacaatt gacattgtga gcggataaca agatactgag cacatcagca
     1201 ggacgcactg accgaattca ttaaagagga gaaaggtacc atggatagca ctgagaacgt
     1261 catcaagccc ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     1321 cgagatcgag ggcgtgggcg agggcaagcc ctacgagggc acccagaccg ccaagctgca
     1381 agtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttcttcta
     1441 cggctccaag gcgtacatca agcaccccgc cgacatcccc gactacctca agcagtcctt
     1501 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     1561 gacccaggac tcctccctgc aggacggcac cctcatctac cacgtgaagt tcatcggcgt
     1621 gaacttcccc tccgacggcc ccgtaatgca gaagaagact ctgggctggg agccctccac
     1681 tgagcgcaac tacccccgcg acggcgtgct gaagggcgag aaccacatgg cgctgaagct
     1741 gaagggcggc ggccactacc tgtgtgagtt caagtccatc tacatggcca agaagcccgt
     1801 gaagctgccc ggctaccact acgtggacta caagctcgac atcacctccc acaacgagga
     1861 ctacaccgtg gtggagcagt acgagcgcgc cgaggcccgc caccacctgt tccagtagtc
     1921 tagaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     1981 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgccctagac ctaggaacaa
     2041 atgcgagatc ttgatgttgg cgtcttcgtt gtcgactgtt cgtcttcgtc ccagtagacg
     2101 tcgatatctg gcgaaaatga gacgttgatc ggcacgtaag aggttccaac tttcaccata
     2161 atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg
     2221 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
     2281 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
     2341 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
     2401 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattccgt atggcaatga
     2461 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
     2521 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
     2581 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
     2641 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
     2701 taaacgtggc caatatggac aacttcttcg c
//
