LOCUS Exported 3348 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pENTRegfp3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3348) AUTHORS Villefranc JA, Amigo J, Lawson ND TITLE Gateway compatible vectors for analysis of gene function in the zebrafish. JOURNAL Dev Dyn. 2007 Nov . 236(11):3077-87. PUBMED 17948311 REFERENCE 2 (bases 1 to 3348) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3348 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 92..109 /label=L4440 /note="L4440 vector, forward primer" terminator 268..295 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 387..473 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 522..544 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 536..553 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 537..553 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 569..668 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" regulatory 671..680 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 677..1393 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 677..1393 /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(722..743) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(983..1002) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1330..1351 /label=EGFP-C /note="EGFP, forward primer" misc_feature 1394..1459 /label=MCS /note="multiple cloning site" protein_bind complement(1473..1572) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1589..1608) /label=T7 /note="T7 promoter, forward primer" promoter complement(1590..1608) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1613..1629) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1613..1629) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1731..1750) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1742..2551 /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(1809..1828) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2698..3286 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3187..3206 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc 241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta 301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc 361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa 421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg 481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa 541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac 601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa 661 agcaggctgg gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat 721 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga 781 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc 841 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta 901 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca 961 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt 1021 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg 1081 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc 1141 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg 1201 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct 1261 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa 1321 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga 1381 cgagctgtac aagtccggac tcagatctcg agctcaagct tcgaattctg cagtcgacgg 1441 taccgcgggc ccgggatcca ccggatctag acacccagct ttcttgtaca aagttggcat 1501 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa 1561 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc 1621 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa 1681 aataatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt 1741 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga 1801 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg 1861 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc 1921 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc 1981 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc 2041 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga 2101 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa 2161 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga 2221 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat 2281 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga 2341 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat 2401 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc 2461 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca 2521 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg 2581 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt 2641 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg 2701 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc 2761 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag 2821 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa 2881 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc 2941 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc 3001 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta 3061 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag 3121 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct 3181 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga 3241 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc 3301 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt //