LOCUS Exported 2942 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pMVP L1-R5 entry plasmid, contains TRE promoter for 4-component MultiSite Gateway Pro assembly. ACCESSION . VERSION . KEYWORDS IG703: pMVP (L1-R5) TRE promoter SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2942) 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 2942) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2942 /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" promoter 674..988 /label=tight TRE promoter /note="Tet-responsive promoter PTight, consisting of seven tet operator sequences followed by the minimal CMV promoter" protein_bind 682..700 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 718..736 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 753..771 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 789..807 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 825..843 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 860..878 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 896..914 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 961..985 /label=LNCX /note="Human CMV promoter, forward primer" protein_bind 1005..1128 /gene="mutant version of attR" /label=attR5 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1183..1202) /label=T7 /note="T7 promoter, forward primer" promoter complement(1184..1202) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1207..1223) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1207..1223) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1325..1344) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1336..2145 /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(1403..1422) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2292..2880 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2781..2800 /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 agcaggctgc ctcgagttta ctccctatca gtgatagaga acgtatgtcg agtttactcc 721 ctatcagtga tagagaacga tgtcgagttt actccctatc agtgatagag aacgtatgtc 781 gagtttactc cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag 841 agaacgtatg tcgagtttat ccctatcagt gatagagaac gtatgtcgag tttactccct 901 atcagtgata gagaacgtat gtcgaggtag gcgtgtacgg tgggaggcct atataagcag 961 agctcgttta gtgaaccgtc agatcgcctg gagaattgga tccacaactt tgtatacaaa 1021 agttgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa 1081 aaaacagact acataatact gtaaaacaca acatatccag tcactatgaa tcaactactt 1141 agatggtatt agtgacctgt actgcagggc ggccgcgata tcccctatag tgagtcgtat 1201 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac 1261 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt 1321 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc 1381 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt 1441 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc 1501 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa 1561 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc 1621 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt 1681 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt 1741 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat 1801 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa 1861 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt 1921 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt 1981 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt 2041 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat 2101 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat 2161 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca 2221 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc 2281 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 2341 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 2401 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta 2461 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 2521 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 2581 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 2641 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 2701 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 2761 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 2821 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 2881 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 2941 tt //