LOCUS Exported 3024 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway entry vector for N-term eGFP fusion, frame C3. ACCESSION . VERSION . KEYWORDS pENTR4-GFP-C3 (w393-1) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3024) AUTHORS Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, Campisi J, Yaswen P, Cooper PK, Kaufman PD. TITLE A versatile viral system for expression and depletion of proteins in mammalian cells. JOURNAL PLoS One. 2009 Aug 6;4(8):e6529. PUBMED 19657394 REFERENCE 2 (bases 1 to 3024) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3024 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 103..189 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 281..308 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" primer_bind 316..339 /label=pENTR-F /note="pENTR vectors, forward primer" protein_bind 358..457 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 463..1179 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 463..1179 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(508..529) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(769..788) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1116..1137 /label=EGFP-C /note="EGFP, forward primer" misc_feature 1185..1241 /label=MCS /note="multiple cloning site" protein_bind complement(1253..1352) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1464..1483) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1475..2284 /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 DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(1542..1561) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2374..2962 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2863..2882 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg 61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct 121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt 181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg 241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat 301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa 361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc 421 ttttttataa tgccaacttt gtacaaaaaa gcaggctcca ccatggtgag caagggcgag 481 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac 541 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag 601 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc 661 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag 721 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac 781 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg 841 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac 901 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc 961 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac 1021 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc 1081 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc 1141 gccgccggga tcactctcgg catggacgag ctgtacaagt actcagatct cgagctcaag 1201 cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agacccagct 1261 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag 1321 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgcagctct ggcccgtgtc 1381 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact 1441 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc 1501 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga 1561 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga 1621 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag 1681 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc 1741 tggtgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga 1801 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt 1861 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca 1921 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa 1981 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga 2041 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt 2101 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat 2161 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata 2221 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt 2281 ctaatcagaa ttggttaatt ggttgtaaca ttattcagat tgggccccgt tccactgagc 2341 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 2401 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 2461 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 2521 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 2581 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 2641 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 2701 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 2761 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 2821 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 2881 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 2941 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt 3001 ttgctggcct tttgctcaca tgtt //