LOCUS       Exported                3445 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Fluorescent reporter for Cre recombinase activity. Cre removes a 
            transcription terminator, leading to RFP production..
ACCESSION   .
VERSION     .
KEYWORDS    pBbAW4k-loxP-TT-loxP-mRFP1
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 May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3445
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(19..96)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     302..321
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     protein_bind    469..502
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     terminator      550..621
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      637..664
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    673..706
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             727..1404
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1425..1496
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1512..1539
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1567..1584)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1701..2246)
                     /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(1781..1800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      2360..2454
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2485..3279)
                     /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(2596..2615)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3206..3225
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 acattaattg cgttgcgcat tcaccaccct gaattgactc tcttccgggc gctatcatgc
       61 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
      121 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
      181 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
      241 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
      301 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
      361 cgatgccgaa gttcctatta tcacgaggcc ctttgagtta tcaaaaagag tattgcatta
      421 aagtctaacc tataggaatc ttacagccat cgagagggac acggcgaaat aacttcgtat
      481 agcatacatt atacgaagtt atccaggcat caaataagga tccaaactcg agtaaggatc
      541 tccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
      601 tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc
      661 tgcgtttata ccataacttc gtatagcata cattatacga agttatttta agaaggagat
      721 atacatatgg cgagtagcga agacgttatc aaagagttca tgcgtttcaa agttcgtatg
      781 gaaggttccg ttaacggtca cgagttcgaa atcgaaggtg aaggtgaagg tcgtccgtac
      841 gaaggtaccc agaccgctaa actgaaagtt accaaaggtg gtccgctgcc gttcgcttgg
      901 gacatcctgt ccccgcagtt ccagtacggt tccaaagctt acgttaaaca cccggctgac
      961 atcccggact acctgaaact gtccttcccg gaaggtttca aatgggaacg tgttatgaac
     1021 ttcgaagacg gtggtgttgt taccgttacc caggactcct ccctgcaaga cggtgagttc
     1081 atctacaaag ttaaactgcg tggtaccaac ttcccgtccg acggtccggt tatgcagaaa
     1141 aaaaccatgg gttgggaagc ttccaccgaa cgtatgtacc cggaagacgg tgctctgaaa
     1201 ggtgaaatca aaatgcgtct gaaactgaaa gacggtggtc actacgacgc tgaagttaaa
     1261 accacctaca tggctaaaaa accggttcag ctgccgggtg cttacaaaac cgacatcaaa
     1321 ctggacatca cctcccacaa cgaagactac accatcgttg aacagtacga acgtgctgaa
     1381 ggtcgtcact ccaccggtgc ttaataatag ggatctccag gcatcaaata aaacgaaagg
     1441 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact
     1501 agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttatacctag ggatatattc
     1561 cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc
     1621 ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag
     1681 agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc
     1741 tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1801 cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg
     1861 ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg
     1921 ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg
     1981 taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac
     2041 tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa
     2101 ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag
     2161 ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga
     2221 ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata aaatatttct
     2281 agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata
     2341 agttgttact agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc
     2401 tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg
     2461 agctctcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
     2521 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
     2581 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
     2641 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
     2701 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag
     2761 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
     2821 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
     2881 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     2941 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     3001 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     3061 cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga
     3121 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     3181 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     3241 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
     3301 ctcttgatca gatcatgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca
     3361 gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca attccgacgt
     3421 cggtgcctaa tgagtgagct aactt
//
