LOCUS       sequence-353901-        3755 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Activatable RFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3755)
  AUTHORS   Tickman BI, Burbano DA, Chavali VP, Kiattisewee C, Fontana J, 
            Khakimzhan A, Noireaux V, Zalatan JG, Carothers JM
  TITLE     Multi-layer CRISPRa/i circuits for dynamic genetic programs in 
            cell-free and bacterial systems.
  JOURNAL   Cell Syst. 2021 Nov 17. pii: S2405-4712(21)00419-1. doi: 
            10.1016/j.cels.2021.10.008.
  PUBMED    34800362
REFERENCE   2  (bases 1 to 3755)
  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: "Cell Syst. 
            2021 Nov 17. pii: S2405-4712(21)00419-1. doi: 
            10.1016/j.cels.2021.10.008."
COMMENT     Sequence Label: pBT009.J3.RFP
FEATURES             Location/Qualifiers
     source          1..3755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     421..440
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(659..763)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     831..849
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1371..1405
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             1415..1426
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1433..2110
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(2623..2640)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2794..3382)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2874..2893)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3553..3755,1..589,590..658))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
       61 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
      121 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
      181 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
      241 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
      301 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
      361 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg
      421 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
      481 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
      541 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
      601 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
      661 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
      721 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
      781 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
      841 tcacgaggcc ctttcgtctt caagaattct ggcgaatcct ctgaccagcc agaaaacgac
      901 ctttctgtgg tgaaaccgga tgctgcaatt cagagcgcca gcaagtgggg gacagcagaa
      961 gacctgaccg ccgcagagtg gatgtttgac atggtgaaga ctatcgcacc atcagccaga
     1021 aaaccgaatt ttgctgggtg ggctaacgat atccgcctga tgcgtgaacg tgacggacgt
     1081 aaccaccgcg acatgtgtgt gctgttccgc tgggcatgcc aggacaactt ctggtccggt
     1141 aacgtgctga gcccggccaa gcttagcccg aaaagtgcca cctgtggcaa ttccgacgtc
     1201 agcatttgcg atcattcacg cagcgcttat tcagttgctc actgcgatgt cataatcatc
     1261 gctacgagct gtgaaagatg cataaagctc gtacgacgcg ttcgctcgtc tcctcacttc
     1321 tcctacggag cgttctggac acaacgtcgt cttgaagttg cgattataga ttgacagcta
     1381 gctcagtcct agggattgtg ctagcgaatt cattaaagag gagaaaggta ccatggcgag
     1441 tagcgaagac gttatcaaag agttcatgcg tttcaaagtt cgtatggaag gttccgttaa
     1501 cggtcacgag ttcgaaatcg aaggtgaagg tgaaggtcgt ccgtacgaag gtacccagac
     1561 cgctaaactg aaagttacca aaggtggtcc gctgccgttc gcttgggaca tcctgtcccc
     1621 gcagttccag tacggttcca aagcttacgt taaacacccg gctgacatcc cggactacct
     1681 gaaactgtcc ttcccggaag gtttcaaatg ggaacgtgtt atgaacttcg aagacggtgg
     1741 tgttgttacc gttacccagg actcctccct gcaagacggt gagttcatct acaaagttaa
     1801 actgcgtggt accaacttcc cgtccgacgg tccggttatg cagaaaaaaa ccatgggttg
     1861 ggaagcttcc accgaacgta tgtacccgga agacggtgct ctgaaaggtg aaatcaaaat
     1921 gcgtctgaaa ctgaaagacg gtggtcacta cgacgctgaa gttaaaacca cctacatggc
     1981 taaaaaaccg gttcagctgc cgggtgctta caaaaccgac atcaaactgg acatcacctc
     2041 ccacaacgaa gactacacca tcgttgaaca gtacgaacgt gctgaaggtc gtcactccac
     2101 cggtgcttaa ggatccaaac tcgagtaagg atctgtgctt tttttaacgc atgagaaagc
     2161 ccccggaaga tcaccttccg ggggcttttt tattgcgcgg atccttactc cgtcgacaag
     2221 cttgcggccg cactcgagga attccgactc aattagttca gtcagtttca ggatattagt
     2281 catctctaca ttgattatga gtattcagaa attccttaaa tattctgaca aatgctcttt
     2341 ccctaaactc cccccataaa aaaacccgcc gaagcgggtt tttacgttat ttgcggatta
     2401 acgattactc gttatcagaa ccgcccagac ctgcgttcag cagttctgcc aggctggcag
     2461 atgcgtcttc cggaattgat ccgtcgaccg atgcccttga gagccttcaa cccagtcagc
     2521 tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc
     2581 atgcaactcg taggacaggt gccggcagcg ctcttccgct tcctcgctca ctgactcgct
     2641 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2701 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2761 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2821 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2881 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2941 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3001 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3061 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3121 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3181 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3241 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3301 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3361 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3421 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3481 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3541 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3601 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3661 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3721 atcagcaata aaccagccag ccggaagggc cgagc
//
