LOCUS Exported 3652 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS 543-p3E-2A-mcherrypA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3652) AUTHORS Villefranc JA, Nicoli S, Bentley K, Jeltsch M, Zarkada G, Moore JC, Gerhardt H, Alitalo K, Lawson ND TITLE A truncation allele in vascular endothelial growth factor c reveals distinct modes of signaling during lymphatic and vascular development. JOURNAL Development. 2013 Apr;140(7):1497-506. doi: 10.1242/dev.084152. Epub 2013 Mar 5. PUBMED 23462469 REFERENCE 2 (bases 1 to 3652) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3652 /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 complement(624..748) /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 759..815 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 816..1526 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(961..979) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1229..1248 /label=mCherry-F /note="mCherry, forward primer" primer_bind 1423..1446 /label=DsRed1-C /note="DsRed1, forward primer" promoter complement(1542..1559) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 1564..1698 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1588..1607) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1642..1661 /label=SV40pA-R /note="SV40 polyA, reverse primer" protein_bind complement(1761..1856) /gene="mutant version of attL" /label=attL3 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1893..1912) /label=T7 /note="T7 promoter, forward primer" promoter complement(1894..1912) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1917..1933) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1917..1933) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2035..2054) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 2046..2855 /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(2113..2132) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 3002..3590 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3491..3510 /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 tcgggccctg cagctctaga gctcgaattc tacaggtcac 601 taataccatc taagtagttg gttcatagtg actgcatatg ttgtgtttta cagtattatg 661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt 721 ctcgttcaac tttcttgtac aaagtggggg gatccggagc cacgaacttc tctctgttaa 781 agcaagcagg agacgtggaa gaaaaccccg gtcctatggt gagcaagggc gaggaggaca 841 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg 901 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg 961 ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccctc 1021 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga 1081 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg 1141 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc 1201 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg 1261 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga 1321 ggctgaagct gaaggacggc ggccactacg acgccgaggt caagaccacc tacaaggcca 1381 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagctggac atcacctccc 1441 acaacgagga ctacaccatc gtggaacagt acgagcgcgc cgagggccgc cactccaccg 1501 gcggcatgga cgagctgtac aagtaaggcg cgcctctaga actatagtga gtcgtattac 1561 gtagatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag 1621 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata 1681 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg 1741 gaggtgtggg aggttttttc caactttatt atacaaagtt ggcattataa aaaagcattg 1801 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttggag 1861 ctccatggta gcgttaacgc ggccgcgata tcccctatag tgagtcgtat tacatggtca 1921 tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag 1981 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg 2041 gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg 2101 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct 2161 atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg 2221 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc 2281 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga 2341 tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg 2401 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt 2461 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg 2521 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag 2581 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac 2641 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg 2701 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc 2761 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat 2821 tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac 2881 actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta 2941 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt 3001 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 3061 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 3121 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact 3181 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 3241 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 3301 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 3361 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 3421 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 3481 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 3541 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 3601 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tt //