LOCUS Exported 3599 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION pMVP L3-L2 entry plasmid, contains mCherry-P2A + polyA for 3- or 4-component MultiSite Gateway Pro assembly. Allows expression of C-term mCherry linked by P2A to gene of interest.. ACCESSION . VERSION . KEYWORDS KY901: pMVP (L3-L2) P2A-mCherry + polyA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3599) AUTHORS Haldeman JM, Conway AE, Arlotto ME, Slentz DH, Muoio DM, Becker TC, Newgard CB TITLE Creation of versatile cloning platforms for transgene expression and dCas9-based epigenome editing. JOURNAL Nucleic Acids Res. 2018 Dec 27. pii: 5264291. doi: 10.1093/nar/gky1286. PUBMED 30590691 REFERENCE 2 (bases 1 to 3599) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3599 /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 570..665 /gene="mutant version of attL" /label=attL3 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 680..736 /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 743..1453 /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(888..906) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1156..1175 /label=mCherry-F /note="mCherry, forward primer" primer_bind complement(1489..1506) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1495..1702 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" protein_bind complement(1711..1810) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1840..1859) /label=T7 /note="T7 promoter, forward primer" promoter complement(1841..1859) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1864..1880) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1864..1880) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1982..2001) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1993..2802 /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(2060..2079) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2949..3537 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3438..3457 /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 tgtataataa 661 agttggtggc ggatccggag ctactaattt ctctctgtta aagcaggcag gagatgtgga 721 ggagaatcca ggtcctaagg gaatggtgag caagggcgag gaggataaca tggccatcat 781 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga 841 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt 901 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg 961 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc 1021 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac 1081 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa 1141 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga 1201 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa 1261 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca 1321 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta 1381 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga 1441 gctgtacaag taagaattct gaagcttagc ctagagctcg ctgatcagcc tcgactgtgc 1501 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag 1561 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta 1621 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag 1681 agaatagcag gcatgctggg gaggtacctg acccagcttt cttgtacaaa gttggcatta 1741 taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa 1801 tcattatttg ccatccagct gcagggcggc cgcgatatcc cctatagtga gtcgtattac 1861 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt 1921 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat 1981 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac 2041 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg 2101 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa 2161 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt 2221 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc 2281 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa 2341 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat 2401 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac 2461 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc 2521 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat 2581 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga 2641 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag 2701 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg 2761 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg 2821 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa 2881 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa 2941 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 3001 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 3061 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 3121 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 3181 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 3241 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 3301 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 3361 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 3421 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 3481 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 3541 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt //