LOCUS       sequence-390452-        2733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses E2-Crimson under the control of PAAH synthetic promoter 
            regulated by AHL and aTc, 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 Dec 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: pO3
FEATURES             Location/Qualifiers
     source          1..2733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(82..168)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(180..857)
                     /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(440..459)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    885..903
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     rep_origin      complement(1169..1714)
                     /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(1249..1268)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1828..1922)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1943..2602)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2517..2536
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2603..2705)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 ctactgggac gaagacgaac agtcgacaac gaagacgcca acatcaagat ctcgcatttg
       61 ttcctaggtc tagggcggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
      121 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctctagac
      181 tactggaaca ggtggtggcg ggcctcggcg cgctcgtact gctccaccac ggtgtagtcc
      241 tcgttgtggg aggtgatgtc gagcttgtag tccacgtagt ggtagccggg cagcttcacg
      301 ggcttcttgg ccatgtagat ggacttgaac tcacacaggt agtggccgcc gcccttcagc
      361 ttcagcgcca tgtggttctc gcccttcagc acgccgtcgc gggggtagtt gcgctcagtg
      421 gagggctccc agcccagagt cttcttctgc attacggggc cgtcggaggg gaagttcacg
      481 ccgatgaact tcacgtggta gatgagggtg ccgtcctgca gggaggagtc ctgggtcacg
      541 gtcaccacgc cgccgtcctc gaagttcatc acgcgctccc acttgaagcc ctcggggaag
      601 gactgcttga ggtagtcggg gatgtcggcg gggtgcttga tgtacgcctt ggagccgtag
      661 aagaactggg gggacaggat gtcccaggcg aagggcaggg ggccgccctt ggtcacttgc
      721 agcttggcgg tctgggtgcc ctcgtagggc ttgccctcgc ccacgccctc gatctcgaac
      781 tcgtggccgt tcacggagcc ctccatgtgc accttgaagc gcatgaaggg cttgatgacg
      841 ttctcagtgc tatccatggt accggactcc tctccaatga attctctcta tcactgatag
      901 ggatttattc gactatactc tatcactgat agggacgtaa acctgtacga tcctacaggt
      961 ctcgagttac caggatcctg gcagaagcta ttcattctgc agttccgtgc tccaaaacaa
     1021 ccgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1081 aatcagggga taacgcagga aagaacatgt ctggaagatg ccaggaagat acttaacagg
     1141 gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct gacaagcatc
     1201 acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg
     1261 cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg
     1321 tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag ttccgggtag
     1381 gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc
     1441 ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc accactggca
     1501 gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta
     1561 aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc ggttcaaaga
     1621 gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga
     1681 gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta atcagataaa
     1741 atatttctag atttcagtgc aatttatctc ttcaaatgta gcacctgaag tcagccccat
     1801 acgatataag ttgttactag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc
     1861 gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag
     1921 tccaagcgag ctcgatatca aattacgccc cgccctgcca ctcatcgcag tactgttgta
     1981 attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg
     2041 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg
     2101 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
     2161 tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
     2221 cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
     2281 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
     2341 cactatccca tatcaccagc tcaccgtctt tcattgccat acggaattcc ggatgagcat
     2401 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
     2461 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     2521 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     2581 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa
     2641 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     2701 gatcaacgtc tcattttcgc cagatatcga cgt
//
