LOCUS       sequence-249605-        2475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EMMA Kit # 1000000119 extension.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2475)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 2475)
  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"
COMMENT     Sequence Label: pRDAX-EMMI-025   eGFP(I-J)//Esp3I
FEATURES             Location/Qualifiers
     source          1..2475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(11..40)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     CDS             join(106..108,109..822)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(106..108,109..819)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(148..169)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(409..428)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     756..777
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      864..895
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        896..998
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             999..1814
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1072..1091)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1838..1865
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1877..2464)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1957..1976)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gattttctac cgaagaaagg cccacccgtg aaggtgagcc agtgagttga ttgcagtcca
       61 gttacgctgg agtctgaggc tcgtcctgaa tgcgtctctg cgtccgtgag caagggcgag
      121 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      181 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      241 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
      301 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
      361 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
      421 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
      481 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
      541 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
      601 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
      661 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
      721 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
      781 gccgccggga tcactctcgg catggacgag ctgtacaagt aatgctagag acgtttaaat
      841 catacctgac ctccatagca gaaagtcaaa agcctccgac cggaggcttt tgacttgatc
      901 ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt
      961 ttgagttatc gagattttca ggagctaagg aagctaaaat gagccatatt caacgggaaa
     1021 cgtcttgctc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     1081 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg
     1141 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     1201 agatggtcag gctaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     1261 tccgtactcc tgatgatgca tggttactca ccactgcgat cccagggaaa acagcattcc
     1321 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     1381 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacggcgat cgcgtatttc
     1441 gcctcgctca ggcgcaatca cgaatgaata acggtttggt tggtgcgagt gattttgatg
     1501 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat
     1561 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     1621 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     1681 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     1741 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcac ttgatgctcg
     1801 atgagttttt ctaatgaggg cccaaatgta atcacctggc tcaccttcgg gtgggccttt
     1861 ctgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgat
     1921 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1981 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2041 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2101 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2161 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2221 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2281 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2341 tgctgaagcc agttacctcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2401 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2461 tcaagaagat ccttt
//
