LOCUS       sequence-314864-        2936 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Contains EGFP flanked by a splice acceptor and a splice donor. 
            Together with other intron tagging plasmids, it can be used to place
            the EGFP tag as a synthetic exon into introns of target genes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2936)
  AUTHORS   Reicher A, Koren A, Kubicek S
  TITLE     Pooled protein tagging, cellular imaging, and in situ sequencing for
            monitoring drug action in real time.
  JOURNAL   Genome Res. 2020 Dec;30(12):1846-1855. doi: 10.1101/gr.261503.120. 
            Epub 2020 Nov 17.
  PUBMED    33203764
REFERENCE   2  (bases 1 to 2936)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1101/gr.261503.120"; journalName: "Genome Res"; date: "2020-12";
            volume: "30"; issue: "12"; pages: "1846-1855"
COMMENT     Sequence Label: Intron-Tagging-EGFP-Donor
FEATURES             Location/Qualifiers
     source          1..2936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(151..153,154..864)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(151..153,154..864)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(193..214)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(454..473)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     801..822
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1009..1031
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1069..1087)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1155..1259
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1260..1328,1329..2120)
                     /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     complement(1478..1497)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2291..2879
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2780..2799
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgttctttc ctgcgttatc ccctggagat cgagtgccgc atcaccggct attggtctta
       61 ctgacatcca ctttgccttt ctctccacag ggtggcggtg gctcgggcgg tggtgggtcc
      121 ggtggcggcg gatctggcgg tggtggatcc gtgagcaagg gcgaggagct gttcaccggg
      181 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
      241 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
      301 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
      361 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
      421 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
      481 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
      541 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
      601 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
      661 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
      721 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
      781 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
      841 ctcggcatgg acgagctgta caagggcggc ggtggatccg gtggcggcgg atctggcggt
      901 ggaggttcgg gtggcggtgg gtcgggcaag gtaagtatca attagaagac gaattcccgg
      961 tgatgcggca ctcgatctcg aattcttaca gacaagctgt gaccgtctcc gggagctgca
     1021 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     1081 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     1141 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     1201 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     1261 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     1321 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     1381 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     1441 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     1501 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     1561 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     1621 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     1681 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     1741 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     1801 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     1861 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     1921 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     1981 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     2041 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     2101 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     2161 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     2221 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2281 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2341 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2401 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     2461 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2521 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     2581 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     2641 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     2701 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     2761 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2821 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     2881 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcac
//
