LOCUS       sequence-390458-        2733 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses E2-Crimson under the control of PIAH synthetic promoter 
            regulated by AHL and IPTG, p15A origin of replication, 
            Chloramphenicol selection,.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2733)
  AUTHORS   Sarkar K, Chakraborty S, Bonnerjee D, Bagh S
  TITLE     Distributed Computing with Engineered Bacteria and Its Application 
            in Solving Chemically Generated 2 x 2 Maze Problems.
  JOURNAL   ACS Synth Biol. 2021 Oct 15;10(10):2456-2464. doi: 
            10.1021/acssynbio.1c00279. Epub 2021 Sep 20.
  PUBMED    34543017
REFERENCE   2  (bases 1 to 2733)
  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"; doi: 
            "10.1021/acssynbio.1c00279"; journalName: "ACS Synth Biol"; date: 
            "2021-10-15- 15"; volume: "10"; issue: "10"; pages: "2456-2464"
COMMENT     Sequence Label: pO6
FEATURES             Location/Qualifiers
     source          1..2733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(81..167)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(179..856)
                     /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     complement(439..458)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     RBS             863..874
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    886..902
                     /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    917..933
                     /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)."
     rep_origin      complement(1168..1713)
                     /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(1248..1267)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1827..1921)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1942..2601)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2516..2535
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2602..2704)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 tactgggacg aagacgaaca gtcgacaacg aagacgccaa catcaagatc tcgcatttgt
       61 tcctaggtct agggcggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag
      121 ataaaacgaa aggcccagtc tttcgactga gcctttcgtt ttatttgatg cctctagact
      181 actggaacag gtggtggcgg gcctcggcgc gctcgtactg ctccaccacg gtgtagtcct
      241 cgttgtggga ggtgatgtcg agcttgtagt ccacgtagtg gtagccgggc agcttcacgg
      301 gcttcttggc catgtagatg gacttgaact cacacaggta gtggccgccg cccttcagct
      361 tcagcgccat gtggttctcg cccttcagca cgccgtcgcg ggggtagttg cgctcagtgg
      421 agggctccca gcccagagtc ttcttctgca ttacggggcc gtcggagggg aagttcacgc
      481 cgatgaactt cacgtggtag atgagggtgc cgtcctgcag ggaggagtcc tgggtcacgg
      541 tcaccacgcc gccgtcctcg aagttcatca cgcgctccca cttgaagccc tcggggaagg
      601 actgcttgag gtagtcgggg atgtcggcgg ggtgcttgat gtacgccttg gagccgtaga
      661 agaactgggg ggacaggatg tcccaggcga agggcagggg gccgcccttg gtcacttgca
      721 gcttggcggt ctgggtgccc tcgtagggct tgccctcgcc cacgccctcg atctcgaact
      781 cgtggccgtt cacggagccc tccatgtgca ccttgaagcg catgaagggc ttgatgacgt
      841 tctcagtgct atccatggta cctttctcct ctttaatgaa ttcaattgtt atccgctcac
      901 aatttattcg actatattgt tatccgctca caaacgtaaa cctgtacgat cctacaggtc
      961 tcgagttacc aggatcctgg cagaagctat tcattctgca gttccgtgct ccaaaacaac
     1021 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     1081 atcaggggat aacgcaggaa agaacatgtc tggaagatgc caggaagata cttaacaggg
     1141 aagtgagagg gccgcggcaa agccgttttt ccataggctc cgcccccctg acaagcatca
     1201 cgaaatctga cgctcaaatc agtggtggcg aaacccgaca ggactataaa gataccaggc
     1261 gtttccccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt
     1321 cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg
     1381 cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct
     1441 tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca ccactggcag
     1501 cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa
     1561 actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg gttcaaagag
     1621 ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc gttttcagag
     1681 caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa tcagataaaa
     1741 tatttctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata
     1801 cgatataagt tgttactagt gcttggattc tcaccaataa aaaacgcccg gcggcaaccg
     1861 agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt
     1921 ccaagcgagc tcgatatcaa attacgcccc gccctgccac tcatcgcagt actgttgtaa
     1981 ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc
     2041 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc
     2101 gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt
     2161 ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc
     2221 gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc
     2281 actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac
     2341 actatcccat atcaccagct caccgtcttt cattgccata cggaattccg gatgagcatt
     2401 catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac
     2461 ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac
     2521 tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata
     2581 tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     2641 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg
     2701 atcaacgtct cattttcgcc agatatcgac gtc
//
