LOCUS       sequence-491314-        3809 bp    DNA     circular SYN 31-OCT-2025
DEFINITION  The plasmid comprises an indicible transcriptional unit encoding 
            EYFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3809)
  AUTHORS   Caliendo F, Vitu E, Wang J, Kuo SH, Sandt H, Enghuus CN, Tordoff J, 
            Estrada N, Collins JJ, Weiss R
  TITLE     Customizable gene sensing and response without altering endogenous 
            coding sequences.
  JOURNAL   Nat Chem Biol. 2024 Sep 12. doi: 10.1038/s41589-024-01733-y.
  PUBMED    39266721
REFERENCE   2  (bases 1 to 3809)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 31, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol. 2024 Sep 12. doi: 10.1038/s41589-024-01733-y."
COMMENT     Sequence Label: pEV749
FEATURES             Location/Qualifiers
     source          1..3809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(262..850)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(342..361)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1021..1812,1813..1881))
                     /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"
     primer_bind     1644..1663
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1882..1986)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      2864..2873
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2870..2872,2873..2875,2876..3589)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2915..2936)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3176..3195)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3523..3544
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3627..3748
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3638..3657)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3692..3711
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 cgcttactga agagcggtct aatacccaat ctctcgtctt atccagatgt tttatacgcc
       61 ttaattaata gggataacag ggtaatatct tgccctgcac gaataccagc gaccccttgc
      121 ccaaatactt gccgtgggcc tcggcctgag agccaaaaca cttgatgcgg aagaagtcgg
      181 tgcgctcctg cttgtcgccg gcatcgttgc gccacatcta ggatctgcca ggaaccgtaa
      241 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      301 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
      361 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
      421 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
      481 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
      541 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
      601 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
      661 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
      721 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
      781 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
      841 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
      901 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
      961 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1021 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     1081 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     1141 cagtgctgca atgataccgc gagaaccacg ctcaccggct ccagatttat cagcaataaa
     1201 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     1261 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     1321 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     1381 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     1441 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     1501 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     1561 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     1621 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     1681 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     1741 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     1801 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     1861 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaagg
     1921 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     1981 tccgcgcacg aattggccag cgctgcatta ccctgttatc cctaaccggt gcgtttttat
     2041 gcttgtagta ttgtataatg tttttaagat ccgctcttca cgcggagttc tgcaattcat
     2101 gtgatctgcg cctgggagct cacggggaca gccccccccc aaagccccca gggatgtaat
     2161 tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct ccggtccggc
     2221 gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg gggaaggtgg
     2281 cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag acacctgggg
     2341 ggatacgggg aaaaagctag gctgaaagag gatcttagaa tgacacgcgc ctgggagctc
     2401 acggggacag ccccccccca aagcccccag ggatgtaatt acgtccctcc cccgctaggg
     2461 ggcagcagcg agccgcccgg ggctccgctc cggtccggcg ctccccccgc atccccgagc
     2521 cggcagcgtg cggggacagc ccgggcacgg ggaaggtggc acgggatcgc tttcctctga
     2581 acgcttctcg ctgctctttg agcctgcaga cacctggggg gatacgggga aaaagctgga
     2641 tcataactcg acgtgcatac tcggagtact gtcctccggc gattaacgga gtactgtcct
     2701 ccggcgatta acggagtact gtcctccggc gattaacgga gtactgtcct ccggcgatta
     2761 acggagttct gtcctccggg gcaccatatg ctctagaggg tatataatgg gggccaaatg
     2821 ctagtctact accagagctc atcgctagcg ctaccggtca ggtgccacca tggtgagcaa
     2881 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     2941 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     3001 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     3061 cttcggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     3121 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     3181 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     3241 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     3301 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     3361 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     3421 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     3481 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     3541 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtgag cttccactgg
     3601 attgtacaat taccaaccag acatgataag atacattgat gagtttggac aaaccacaac
     3661 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     3721 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     3781 ggttcagggg gaggtgtggg aggtttttt
//
