LOCUS Exported 3277 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pENTR-L5-EGFP-L2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3277) AUTHORS White MD, Milne RV, Nolan MF TITLE A Molecular Toolbox for Rapid Generation of Viral Vectors to Up- or Down-Regulate Neuronal Gene Expression in vivo. JOURNAL Front Mol Neurosci. 2011;4:8. Epub 2011 Jul 4. PUBMED 21772812 REFERENCE 2 (bases 1 to 3277) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3277 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(119..138) /label=T7 /note="T7 promoter, forward primer" promoter complement(120..138) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(143..159) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(143..159) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(261..280) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 272..1081 /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 complement(339..358) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 1228..1816 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1717..1736 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1970..1987 /label=L4440 /note="L4440 vector, forward primer" terminator 2146..2173 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 2265..2351 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 2400..2422 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2414..2431 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2415..2431 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 2448..2543 /gene="mutant version of attL" /label=attL5 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 2546..3265 /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 2546..3262 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2591..2612) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2852..2871) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3199..3220 /label=EGFP-C /note="EGFP, forward primer" protein_bind complement(3267..89) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" ORIGIN 1 ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc 61 actatcagtc aaaataaaat cattatttgc catccagctg cagggcggcc gcgatatccc 121 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca 181 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc 241 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag 301 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa 361 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt 421 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact 481 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga 541 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga 601 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca 661 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc 721 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg 781 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc 841 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat 901 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct 961 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg 1021 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta 1081 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg 1141 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga 1201 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 1261 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 1321 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct 1381 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 1441 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 1501 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 1561 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 1621 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 1681 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 1741 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 1801 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 1861 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac 1921 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt 1981 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat 2041 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc 2101 aattaatacg cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc 2161 aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac 2221 cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg 2281 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt 2341 cgttttattt gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc 2401 ccagtcacga cgttgtaaaa cgacggccag tcttaagctc gggccccaaa taatgatttt 2461 attttgactg atagtgacct gttcgttgca acaaattgat gagcaatgct tttttataat 2521 gccaactttg tatacaaaag ttgtgatggt gagcaagggc gaggagctgt tcaccggggt 2581 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg 2641 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg 2701 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt 2761 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg 2821 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga 2881 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa 2941 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta 3001 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat 3061 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg 3121 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc 3181 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct 3241 cggcatggac gagctgtaca agtaataccc agctttc //