LOCUS Exported 3384 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pMVP R4-R3 entry plasmid, contains eYFP (no stop codon) for 3- or 4-component MultiSite Gateway Pro assembly. ACCESSION . VERSION . KEYWORDS IO401: pMVP (R4-R3) eYFP open SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3384) 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 3384) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3384 /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(603..727) /gene="mutant version of attR" /label=attR4 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 729..1445 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(774..795) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1035..1054) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1382..1403 /label=EGFP-C /note="EGFP, forward primer" protein_bind 1447..1570 /gene="mutant version of attR" /label=attR3 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1625..1644) /label=T7 /note="T7 promoter, forward primer" promoter complement(1626..1644) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1649..1665) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1649..1665) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1767..1786) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1778..2587 /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(1845..1864) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2734..3322 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3223..3242 /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 tcgggcccct acaggtcact aataccatct aagtagttga 601 ttcatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa 661 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcaact tttctataca 721 aagttgccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg 781 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg 841 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct 901 ggcccaccct cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc taccccgacc 961 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca 1021 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg 1081 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc 1141 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc 1201 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc 1261 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg 1321 acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc 1381 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt 1441 acaagacaac tttgtataat aaagttgaac gagaaacgta aaatgatata aatatcaata 1501 tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatcc 1561 agtcactatg aatcaactac ttagatggta ttagtgacct gtactgcagg gcggccgcga 1621 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt 1681 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa 1741 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac 1801 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg 1861 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc 1921 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt 1981 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac 2041 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt 2101 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg 2161 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc 2221 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg 2281 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc 2341 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa 2401 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc 2461 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa 2521 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt 2581 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg 2641 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc 2701 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 2761 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 2821 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 2881 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 2941 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 3001 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 3061 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 3121 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 3181 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 3241 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 3301 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt 3361 ttgctggcct tttgctcaca tgtt //