LOCUS       sequence-264478-        3445 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for Cre recombinase activity. Cre removes a 
            transcription terminator, leading to RFP production..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3445)
  AUTHORS   Sheets MB, Wong WW, Dunlop MJ
  TITLE     Light-Inducible Recombinases for Bacterial Optogenetics.
  JOURNAL   ACS Synth Biol. 2020 Feb 21;9(2):227-235. doi: 
            10.1021/acssynbio.9b00395. Epub 2020 Jan 21.
  PUBMED    31961670
REFERENCE   2  (bases 1 to 3445)
  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.9b00395"; journalName: "ACS Synth Biol"; date: 
            "2020-02-21- 21"; volume: "9"; issue: "2"; pages: "227-235"
COMMENT     Sequence Label: pBbAW4k-loxP-TT-loxP-mRFP1
FEATURES             Location/Qualifiers
     source          1..3445
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(29..106)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     312..331
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     protein_bind    complement(479..512)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      560..631
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      647..674
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    complement(683..716)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             737..1414
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1435..1506
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1522..1549
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1577..1594)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1711..2256)
                     /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(1791..1810)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2370..2464)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2495..3289)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2606..2625)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3216..3235
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gagctaactt acattaattg cgttgcgcat tcaccaccct gaattgactc tcttccgggc
       61 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc
      121 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
      181 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
      241 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
      301 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
      361 atgccggcca cgatgccgaa gttcctatta tcacgaggcc ctttgagtta tcaaaaagag
      421 tattgcatta aagtctaacc tataggaatc ttacagccat cgagagggac acggcgaaat
      481 aacttcgtat agcatacatt atacgaagtt atccaggcat caaataagga tccaaactcg
      541 agtaaggatc tccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
      601 ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg
      661 tgggcctttc tgcgtttata ccataacttc gtatagcata cattatacga agttatttta
      721 agaaggagat atacatatgg cgagtagcga agacgttatc aaagagttca tgcgtttcaa
      781 agttcgtatg gaaggttccg ttaacggtca cgagttcgaa atcgaaggtg aaggtgaagg
      841 tcgtccgtac gaaggtaccc agaccgctaa actgaaagtt accaaaggtg gtccgctgcc
      901 gttcgcttgg gacatcctgt ccccgcagtt ccagtacggt tccaaagctt acgttaaaca
      961 cccggctgac atcccggact acctgaaact gtccttcccg gaaggtttca aatgggaacg
     1021 tgttatgaac ttcgaagacg gtggtgttgt taccgttacc caggactcct ccctgcaaga
     1081 cggtgagttc atctacaaag ttaaactgcg tggtaccaac ttcccgtccg acggtccggt
     1141 tatgcagaaa aaaaccatgg gttgggaagc ttccaccgaa cgtatgtacc cggaagacgg
     1201 tgctctgaaa ggtgaaatca aaatgcgtct gaaactgaaa gacggtggtc actacgacgc
     1261 tgaagttaaa accacctaca tggctaaaaa accggttcag ctgccgggtg cttacaaaac
     1321 cgacatcaaa ctggacatca cctcccacaa cgaagactac accatcgttg aacagtacga
     1381 acgtgctgaa ggtcgtcact ccaccggtgc ttaataatag ggatctccag gcatcaaata
     1441 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     1501 gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttatacctag
     1561 ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag
     1621 cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca
     1681 gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca
     1741 tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca
     1801 ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg
     1861 tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt
     1921 aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg
     1981 ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg
     2041 cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc
     2101 taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa
     2161 gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca
     2221 gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata
     2281 aaatatttct agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc
     2341 atacgatata agttgttact agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa
     2401 ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg
     2461 agtccaagcg agctctcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat
     2521 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
     2581 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
     2641 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
     2701 tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc
     2761 gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
     2821 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
     2881 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
     2941 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
     3001 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
     3061 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca
     3121 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
     3181 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
     3241 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc
     3301 ctcatcctgt ctcttgatca gatcatgatc ccctgcgcca tcagatcctt ggcggcaaga
     3361 aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca
     3421 attccgacgt cggtgcctaa tgagt
//
