LOCUS       sequence-492309-        3769 bp    DNA     circular SYN 07-NOV-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 3769)
  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 3769)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 7, 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: pEV712
FEATURES             Location/Qualifiers
     source          1..3769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(247..835)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(327..346)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1006..1797,1798..1866))
                     /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     1629..1648
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1867..1971)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2710..2772
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     regulatory      2820..2829
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2826..2828,2829..2831,2832..3545)
                     /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(2871..2892)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3132..3151)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3479..3500
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3583..3704
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3594..3613)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3648..3667
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ggtgcgtttt tatgcttgta gtattgtata atgtttttaa gatccttaat taatagggat
       61 aacagggtaa tatcttgccc tgcacgaata ccagcgaccc cttgcccaaa tacttgccgt
      121 gggcctcggc ctgagagcca aaacacttga tgcggaagaa gtcggtgcgc tcctgcttgt
      181 cgccggcatc gttgcgccac atctaggatc tgccaggaac cgtaaaaagg ccgcgttgct
      241 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      301 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      361 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      421 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      481 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      541 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      601 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      661 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
      721 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
      781 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
      841 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
      901 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
      961 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1021 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1081 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1141 accgcgagaa ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1201 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1261 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1321 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1381 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1441 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1501 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1561 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     1621 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     1681 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     1741 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     1801 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     1861 actcatactc ttcctttttc aatattattg aagcatttat caaggttatt gtctcatgag
     1921 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacgaattg
     1981 gccagcgctg cattaccctg ttatccctaa ccggtttacc gagctcttat tggttttcaa
     2041 acttcattga ctgtgccgga gttctgcaat tcatgtgatc tgcgcctggg agctcacggg
     2101 gacagccccc ccccaaagcc cccagggatg taattacgtc cctcccccgc tagggggcag
     2161 cagcgagccg cccggggctc cgctccggtc cggcgctccc cccgcatccc cgagccggca
     2221 gcgtgcgggg acagcccggg cacggggaag gtggcacggg atcgctttcc tctgaacgct
     2281 tctcgctgct ctttgagcct gcagacacct ggggggatac ggggaaaaag ctaggctgaa
     2341 agaggatctt agaatgacac gcgcctggga gctcacgggg acagcccccc cccaaagccc
     2401 ccagggatgt aattacgtcc ctcccccgct agggggcagc agcgagccgc ccggggctcc
     2461 gctccggtcc ggcgctcccc ccgcatcccc gagccggcag cgtgcgggga cagcccgggc
     2521 acggggaagg tggcacggga tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg
     2581 cagacacctg gggggatacg gggaaaaagc tggatcataa ctcgacgtgc atactgtccc
     2641 ctccacccca cagtggggca tagcgccggc cgcgccgtcc cctccacccc acagtggggc
     2701 accatatgct tcgcatatta aggtgacgcg tgtggcctcg aacaccgagc gaccctgcag
     2761 cgacccgctt aaaatgctag tctactacca gagctcatcg ctagcgctac cggtcaggtg
     2821 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     2881 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     2941 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     3001 ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac cccgaccaca
     3061 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     3121 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     3181 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     3241 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     3301 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     3361 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3421 accactacct gagctaccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca
     3481 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3541 agtgagcttc cactggattg tacaattacc aaccagacat gataagatac attgatgagt
     3601 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3661 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     3721 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttcgct
//
