LOCUS       sequence-8944-ex        2607 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2607)
  AUTHORS   Venken KJ, Kasprowicz J, Kuenen S, Yan J, Hassan BA, Verstreken P
  TITLE     Recombineering-mediated tagging of Drosophila genomic constructs for
            in vivo localization and acute protein inactivation.
  JOURNAL   Nucleic Acids Res. 2008 Oct;36(18):e114. doi: 10.1093/nar/gkn486. 
            Epub 2008 Aug 1.
  PUBMED    18676454
REFERENCE   2  (bases 1 to 2607)
  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: "Nucleic 
            Acids Res."; date: "2008-10"; volume: "36(18):e114. doi"; pages: " 
            10.1093/nar/gkn486. Epub 2008 Aug 1"
COMMENT     Sequence Label: PL-452 C-EGFP
FEATURES             Location/Qualifiers
     source          1..2607
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    37..70
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        135..634
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(170..193)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     567..586
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        646..693
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             712..1515
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(775..794)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1385..1404
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1547..1564)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1553..1777
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    1834..1867
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             join(1888..1890,1891..1893,1894..2607)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1888..1890,1891..1893,1894..2604)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1933..1954)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2194..2213)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2541..2562
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gcagcccaat tccgatcata ttcaataacc cttaatataa cttcgtataa tgtatgctat
       61 acgaagttat taggtctgaa gaggagttta cgtccagcca agctagcttg gctgcaggtc
      121 gtcgaaattc taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt
      181 agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac
      241 atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac
      301 tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca
      361 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa
      421 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca
      481 gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg
      541 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc
      601 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agcctgttga caattaatca
      661 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac catgggatcg
      721 gccattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
      781 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
      841 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
      901 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
      961 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     1021 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     1081 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     1141 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     1201 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     1261 ggcgatgatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     1321 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     1381 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     1441 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     1501 gacgagttct tctgagggga tcaattctct agagctcgct gatcagcctc gactgtgcct
     1561 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     1621 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     1681 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     1741 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     1801 tggggctcga ctagagcttg cggaaccctt aatataactt cgtataatgt atgctatacg
     1861 aagttattag gtccctcgag gggatccatg gtgagcaagg gcgaggagct gttcaccggg
     1921 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1981 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     2041 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     2101 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     2161 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     2221 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     2281 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     2341 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     2401 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     2461 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     2521 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     2581 ctcggcatgg acgagctgta caagtaa
//
