LOCUS       sequence-409068-        3603 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EL222_D7_mRFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3603)
  AUTHORS   Alba Burbano D, Cardiff RAL, Tickman BI, Kiattisewee C, Maranas CJ, 
            Zalatan JG, Carothers JM
  TITLE     Engineering activatable promoters for scalable and multi-input 
            CRISPRa/i circuits.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Jul 25;120(30):e2220358120. doi: 
            10.1073/pnas.2220358120. Epub 2023 Jul 18.
  PUBMED    37463216
REFERENCE   2  (bases 1 to 3603)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2023-07-25"; volume: "120(30):e2220358120. 
            doi"; pages: " 10.1073/pnas.2220358120. Epub 2023 Jul 18"
COMMENT     Sequence Label: pDA040.BLD7-mRFP
FEATURES             Location/Qualifiers
     source          1..3603
                     /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"
     RBS             1263..1274
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1281..1958
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(2471..2488)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2642..3230)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2722..2741)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3401..3603,1..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 ggtagccttt agtccatgtt gacggtgaag agtatcagag ggattgtgct agcgaattca
     1261 ttaaagagga gaaaggtacc atggcgagta gcgaagacgt tatcaaagag ttcatgcgtt
     1321 tcaaagttcg tatggaaggt tccgttaacg gtcacgagtt cgaaatcgaa ggtgaaggtg
     1381 aaggtcgtcc gtacgaaggt acccagaccg ctaaactgaa agttaccaaa ggtggtccgc
     1441 tgccgttcgc ttgggacatc ctgtccccgc agttccagta cggttccaaa gcttacgtta
     1501 aacacccggc tgacatcccg gactacctga aactgtcctt cccggaaggt ttcaaatggg
     1561 aacgtgttat gaacttcgaa gacggtggtg ttgttaccgt tacccaggac tcctccctgc
     1621 aagacggtga gttcatctac aaagttaaac tgcgtggtac caacttcccg tccgacggtc
     1681 cggttatgca gaaaaaaacc atgggttggg aagcttccac cgaacgtatg tacccggaag
     1741 acggtgctct gaaaggtgaa atcaaaatgc gtctgaaact gaaagacggt ggtcactacg
     1801 acgctgaagt taaaaccacc tacatggcta aaaaaccggt tcagctgccg ggtgcttaca
     1861 aaaccgacat caaactggac atcacctccc acaacgaaga ctacaccatc gttgaacagt
     1921 acgaacgtgc tgaaggtcgt cactccaccg gtgcttaagg atccaaactc gagtaaggat
     1981 ctgtgctttt tttaacgcat gagaaagccc ccggaagatc accttccggg ggctttttta
     2041 ttgcgcggat ccttactccg tcgacaagct tgcggccgca ctcgaggaat tccgactcaa
     2101 ttagttcagt cagtttcagg atattagtca tctctacatt gattatgagt attcagaaat
     2161 tccttaaata ttctgacaaa tgctctttcc ctaaactccc cccataaaaa aacccgccga
     2221 agcgggtttt tacgttattt gcggattaac gattactcgt tatcagaacc gcccagacct
     2281 gcgttcagca gttctgccag gctggcagat gcgtcttccg gaattgatcc gtcgaccgat
     2341 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     2401 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     2461 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2521 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2581 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2641 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2701 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2761 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2821 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2881 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2941 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3001 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3061 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3121 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3181 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3241 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3301 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3361 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3421 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3481 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3541 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3601 agc
//
