LOCUS       sequence-332577-        6161 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the synthetic gene BLADE FP6 under the constitutive 
            J23101* promoter. mCherry expression is controlled by the PBAD 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6161)
  AUTHORS   Romano E, Baumschlager A, Akmeric EB, Palanisamy N, Houmani M, 
            Schmidt G, Ozturk MA, Ernst L, Khammash M, Di Ventura B
  TITLE     Engineering AraC to make it responsive to light instead of 
            arabinose.
  JOURNAL   Nat Chem Biol. 2021 Apr 26. pii: 10.1038/s41589-021-00787-6. doi: 
            10.1038/s41589-021-00787-6.
  PUBMED    33903769
REFERENCE   2  (bases 1 to 6161)
  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"; journalName: "Nat Chem 
            Biol. 2021 Apr 26. pii: 10.1038/s41589-021-00787-6. doi: 
            10.1038/s41589-021-00787-6."
COMMENT     Sequence Label: pBLADE(FP6*)-mCherry
FEATURES             Location/Qualifiers
     source          1..6161
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      49..76
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        95..186
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      644..1099
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(731..750)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     941..962
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1655..2314)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2229..2248
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2315..2417)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2809..2826)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2943..3488)
                     /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(3023..3042)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3564..3583
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        4736..5020
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     4943..4962
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             5074..5784
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5219..5237)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5487..5506
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(5862..5879)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      6032..6118
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc
       61 tgacggatgg cctttttgcg tttctacaaa ctcttttgtt tatttttcta aatacattca
      121 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
      181 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
      241 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
      301 ggtgcagcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
      361 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
      421 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
      481 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
      541 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
      601 aactgtcaga ccaagtttac tcatatatac tttagattga tttacgcgcc ctgtagcggc
      661 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
      721 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
      781 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
      841 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
      901 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
      961 ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     1021 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     1081 atattaacgt ttacaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1141 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1201 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1261 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1321 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     1381 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1441 agtcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac
     1501 cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac cgggtcgaat
     1561 ttgctttcga atttctgcca ttcatccgct tattatcact tattcaggcg tagcaaccag
     1621 gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc
     1681 agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat
     1741 gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg
     1801 tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac
     1861 tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg
     1921 ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat
     1981 cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt
     2041 aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc atacggaatt
     2101 ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct
     2161 tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg
     2221 tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat
     2281 caacggtggt atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc
     2341 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
     2401 ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt ggcccagggc ttcccggtat
     2461 caacagggac accaggattt atttattctg cgaagtgatc ttccgtcaca ggtatttatt
     2521 cggcgcaaag tgcgtcgggt gatgctgcca acttactgat ttagtgtatg atggtgtttt
     2581 tgaggtgctc cagtggcttc tgtttctatc agctgtccct cctgttcagc tactgacggg
     2641 gtggtgcgta acggcaaaag caccgccgga catcagcgct agcggagtgt atactggctt
     2701 actatgttgg cactgatgag ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc
     2761 tgcaccggtg cgtcagcaga atatgtgata caggatatat tccgcttcct cgctcactga
     2821 ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga
     2881 tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg
     2941 tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg
     3001 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg
     3061 cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg
     3121 tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat
     3181 gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3241 caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg
     3301 agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact
     3361 gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa
     3421 aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac
     3481 gatctcaaga agatcatctt attaatcaga taaaatattt ctaggctcat gagcccgaag
     3541 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     3601 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatctgctc atgtttgaca
     3661 gcttatcatt ttttaggcta ccgatgtagc ctaacgcgat ttatgacaac ttgacggcta
     3721 catcattcac tttttcttca caaccggcac ggaactcgct tgggctggcc ccggtgcatt
     3781 ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg
     3841 cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc
     3901 gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg
     3961 acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc
     4021 tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgtttt
     4081 ccgtttcgca ctggaaaccc atgctgtacc ggtattcccc tgtttcatct cgcaccggaa
     4141 tcatcgtcaa gaagttgaca aaccgttggc cgttcttctt aaaattgacc acctcaacct
     4201 gcacctcggc gttcctatca atcgctttcc tcatcgtatt gatcgtgttg gagtcgacgt
     4261 acttccttgt cgatttcggc ttgaccattc cgtcgggtga ctgaagaaaa cggcagtttc
     4321 tccccaagac ctccgcattg ctgtatcctg tcatatagag aaaagcttcc gaggcgtaca
     4381 caattggcgt gtctttttgc ttcaggtcgc acagaatcag agcaactgac gtgtcaacag
     4441 gtcccagttc tacttggggg tttggcctct tcataatctg aatcagatag cccataatgt
     4501 cataaccgcc aggagcgtag agcgtatgca tggtatatct ccttctctag tctctagtag
     4561 ctagcatata cctaggactg agctagctgt aaactctaga atgcataatg tgcctgtcaa
     4621 atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc aattgtctga
     4681 ttcgttacca acggatgcgc catgacagga cttttcatac tcccgccatt cagagaagaa
     4741 accaattgtc catattgcat cagacattgc cgtcactgcg tcttttactg gctcttctcg
     4801 ctaaccaaac cggtaacccc gcttattaaa agcattctgt aacaaagcgg gaccaaagcc
     4861 atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca cattgattat
     4921 ttgcacggcg tcacactttg ctatgccata gcatttttat ccataagatt agcggatcct
     4981 acctgacgct ttttatcgca actctctact gtttctccat acccgttttt tgggctagcg
     5041 aattcgagct cggtacctaa ggaggattta aagatggtga gcaagggcga ggaggataac
     5101 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     5161 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     5221 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     5281 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     5341 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     5401 gtgaccgtga cccaggactc ctccttgcag gacggcgagt tcatctacaa ggtgaagctg
     5461 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     5521 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     5581 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     5641 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
     5701 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     5761 ggcatggacg agctgtacaa gtaaaagctt ggctgttttg gcatggacga gctgtacaag
     5821 taaaagcttg gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca
     5881 gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca
     5941 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct
     6001 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     6061 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     6121 gccgggagcg gatttgaacg ttgcgaagca acggcccgga g
//
