LOCUS       sequence-364493-        3300 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector containing EGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3300)
  AUTHORS   Yang X, Boehm JS, Yang X, Salehi-Ashtiani K, Hao T, Shen Y, Lubonja 
            R, Thomas SR, Alkan O, Bhimdi T, Green TM, Johannessen CM, Silver 
            SJ, Nguyen C, Murray RR, Hieronymus H, Balcha D, Fan C, Lin C, 
            Ghamsari L, Vidal M, Hahn WC, Hill DE, Root DE
  TITLE     A public genome-scale lentiviral expression library of human ORFs.
  JOURNAL   Nat Methods. 2011 Jun 26;8(8):659-61. doi: 10.1038/nmeth.1638
  PUBMED    21706014
REFERENCE   2  (bases 1 to 3300)
  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"; doi: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2011-06-26- 26"; volume: "8"; 
            issue: "8"; pages: "659-61"
COMMENT     Sequence Label: pDONR221_EGFP
FEATURES             Location/Qualifiers
     source          1..3300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     42..63
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(129..228)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(244..261)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(244..260)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(253..275)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      324..410
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      502..529
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(688..705)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(859..1447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(939..958)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1594..2403)
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2317..2336
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2395..2414
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2516..2532
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2516..2532
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2537..2556
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2537..2555
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2573..2672
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(join(2692..3300,1..102,103..105,106..108))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2734..2755)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3083..3102
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 gccctcgccc tcgccagaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac
       61 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg tgaagggggc
      121 ggccgcggag cctgcttttt tgtacaaagt tggcattata aaaaagcatt gctcatcaat
      181 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttggg gcccgagctt
      241 aagactggcc gtcgttttac aacgtcgtga ctgggaaaac atccatgcta gcgttaacgc
      301 gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcggaa gactgggcct
      361 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag
      421 cggatttgaa cgttgtgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa
      481 ctgccaggca tcaaactaag cagaaggcca tcctgacgga tggccttttt gcgtttctac
      541 aaactcttcc tggctagcgg tacgcgtatt aattgcgttg cgctcactgc ccgctttcca
      601 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
      661 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
      721 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      781 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      841 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      901 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      961 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1021 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1081 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1141 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1201 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1261 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1321 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1381 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1441 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgacgcgta
     1501 actcacgtta agggattttg gtcatgagct tgcgccgtcc cgtcaagtca gcgtaatgct
     1561 ctgccagtgt tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg
     1621 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     1681 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     1741 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     1801 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt
     1861 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     1921 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     1981 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     2041 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     2101 tccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     2161 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     2221 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     2281 caagcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     2341 taaatcagca tccatgttgg aatttaatcg cggcctcgac gtttcccgtt gaatatggct
     2401 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat
     2461 atttttatct tgtgcaatgt aacatcagag attttgagac acgggccaga gctgccagga
     2521 aacagctatg accatgtaat acgactcact ataggggata tcagctggat ggcaaataat
     2581 gattttattt tgactgatag tgacctgttc gttgcaacaa attgataagc aatgctttct
     2641 tataatgcca actttgtaca agaaagctgg gtcggcgcgc ccacccttgg acttgtacag
     2701 ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc
     2761 gcgcttctcg ttggggtctt tgctcagggc ggactgggtg ctcaggtagt ggttgtcggg
     2821 cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac
     2881 gctgccgtcc tcgatgttgt ggcggatctt gaagttcgcc ttgatgccgt tcttctgctt
     2941 gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat
     3001 gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc
     3061 ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg
     3121 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc
     3181 ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg
     3241 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc
//
