LOCUS Exported 3364 bp ds-DNA circular SYN 09-DEC-2022 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pENTREGFP2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3364) 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 3364) AUTHORS . TITLE Direct Submission JOURNAL Exported Dec 9, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3364 /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 680..689 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 686..1402 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 686..1402 /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(731..752) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(992..1011) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1339..1360 /label=EGFP-C /note="EGFP, forward primer" misc_feature 1403..1468 /label=MCS /note="multiple cloning site" protein_bind complement(1489..1588) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1605..1624) /label=T7 /note="T7 promoter, forward primer" promoter complement(1606..1624) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1629..1645) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1629..1645) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1747..1766) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1758..2567 /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(1825..1844) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2714..3302 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3203..3222 /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 caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa 661 agcaggctcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt 721 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg 781 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg 841 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt 901 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg 961 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga 1021 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa 1081 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta 1141 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat 1201 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg 1261 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc 1321 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct 1381 cggcatggac gagctgtaca agtccggact cagatctcga gctcaagctt cgaattctgc 1441 agtcgacggt accgcgggcc cgggatccac cggatctaga taactgatac ccagctttct 1501 tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca 1561 ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt 1621 attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt 1681 acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca 1741 gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt 1801 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag 1861 gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg 1921 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg 1981 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac 2041 tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag 2101 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt 2161 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga 2221 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac 2281 aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg 2341 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg 2401 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg 2461 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg 2521 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta 2581 attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac 2641 caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga 2701 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 2761 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 2821 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt 2881 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 2941 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 3001 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 3061 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 3121 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 3181 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 3241 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 3301 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 3361 tgtt //