LOCUS       sequence-390459-        3507 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tdTomato 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 3507)
  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 3507)
  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: pO5
FEATURES             Location/Qualifiers
     source          1..3507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    86..102
                     /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    117..133
                     /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             145..156
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     regulatory      167..176
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             173..1603
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     terminator      1626..1712
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        1822..1924
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1925..2584
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1991..2010)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2605..2699
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2813..3358
                     /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     3259..3278
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgttttggag cacggaactg cagaatgaat agcttctgcc aggatcctgg taactcgaga
       61 cctgtaggat cgtacaggtt tacgtttgtg agcggataac aatatagtcg aataaattgt
      121 gagcggataa caattgaatt cattaaagag gagaaaggta ccggtcgcca ccatggtgag
      181 caagggcgag gaggtcatca aagagttcat gcgcttcaag gtgcgcatgg agggctccat
      241 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
      301 gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc
      361 cccccagttc atgtacggct ccaaggcgta cgtgaagcac cccgccgaca tccccgatta
      421 caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
      481 cggtctggtg accgtgaccc aggactcctc cctgcaggac ggcacgctga tctacaaggt
      541 gaagatgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga agaccatggg
      601 ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca
      661 ccaggccctg aagctgaagg acggcggcca ctacctggtg gagttcaaga ccatctacat
      721 ggccaagaag cccgtgcaac tgcccggcta ctactacgtg gacaccaagc tggacatcac
      781 ctcccacaac gaggactaca ccatcgtgga acagtacgag cgctccgagg gccgccacca
      841 cctgttcctg gggcatggca ccggcagcac cggcagcggc agctccggca ccgcctcctc
      901 cgaggacaac aacatggccg tcatcaaaga gttcatgcgc ttcaaggtgc gcatggaggg
      961 ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     1021 cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat
     1081 cctgtccccc cagttcatgt acggctccaa ggcgtacgtg aagcaccccg ccgacatccc
     1141 cgattacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     1201 ggacggcggt ctggtgaccg tgacccagga ctcctccctg caggacggca cgctgatcta
     1261 caaggtgaag atgcgcggca ccaacttccc ccccgacggc cccgtaatgc agaagaagac
     1321 catgggctgg gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga
     1381 gatccaccag gccctgaagc tgaaggacgg cggccactac ctggtggagt tcaagaccat
     1441 ctacatggcc aagaagcccg tgcaactgcc cggctactac tacgtggaca ccaagctgga
     1501 catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgct ccgagggccg
     1561 ccaccacctg ttcctgtacg gcatggacga gctgtacaag taggcggccg cgactctaga
     1621 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     1681 tgtcggtgaa cgctctcctg agtaggacaa atccgccgcc ctagacctag gaacaaatgc
     1741 gagatcttga tgttggcgtc ttcgttgtcg actgttcgtc ttcgtcccag tagacgtcga
     1801 tatctggcga aaatgagacg ttgatcggca cgtaagaggt tccaactttc accataatga
     1861 aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag ctaaggaagc
     1921 taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa
     1981 agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct
     2041 ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt
     2101 tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga
     2161 cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac
     2221 tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat
     2281 atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat
     2341 tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa
     2401 cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca
     2461 aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt
     2521 ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc
     2581 gtaatttgat atcgagctcg cttggactcc tgttgataga tccagtaatg acctcagaac
     2641 tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc
     2701 caagcactag taacaactta tatcgtatgg ggctgacttc aggtgctaca tttgaagaga
     2761 taaattgcac tgaaatctag aaatatttta tctgattaat aagatgatct tcttgagatc
     2821 gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt
     2881 tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt ggaggagcgc
     2941 agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc aagactaact
     3001 cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg
     3061 gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg gggttcgtgc
     3121 atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg tggaatgaga
     3181 caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa caggagagcg
     3241 cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3301 ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa
     3361 cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc ttccagacat
     3421 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     3481 tgataccgct cgccgcagcc gaacggt
//
