LOCUS       sequence-344981-        3753 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 3753)
  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 3753)
  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..3753
                     /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(2621..2638)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2792..3380)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2872..2891)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3551..3753,1..589,590..658))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     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 tcaatacggg
      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 cagagcggca 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 attcgactca attagttcag tcagtttcag gatattagtc
     2281 atctctacat tgattatgag tattcagaaa ttccttaaat attctgacaa atgctctttc
     2341 cctaaactcc ccccataaaa aaacccgccg aagcgggttt ttacgttatt tgcggattaa
     2401 cgattactcg ttatcagaac cgcccagacc tgcgttcagc agttctgcca ggctggcaga
     2461 tgcgtcttcc gaattgatcc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc
     2521 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     2581 gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc
     2641 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2701 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2761 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2821 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2881 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2941 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     3001 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3061 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3121 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3181 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     3241 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3301 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3361 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3421 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3481 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3541 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3601 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3661 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3721 cagcaataaa ccagccagcc ggaagggccg agc
//
