LOCUS Exported 5722 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pMVP R4-R3 entry plasmid, contains beta-gal for 3- or 4-component MultiSite Gateway Pro assembly. ACCESSION . VERSION . KEYWORDS HD301: pMVP (R4-R3) beta-gal SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5722) 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 5722) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5722 /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 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 735..3782 /codon_start=1 /gene="lacZ" /product="beta-galactosidase" /label=lacZ /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS LNGEWRFAWFPAPEAVPESWLECDLPEADTVVAPSNWQMHGYDAPIYSNVTYPITVNPP FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ PNATAWSETGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA ASLQCTADTLADAALITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP ARIGLNCQLAQVAERVNWLGLGPQENYPDRPTAACFDRWDLPLSDMYTPYVFPSENGLR CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD SWSPSVSAEFQLSAGRYHYQLVWCQK" protein_bind 3785..3908 /gene="mutant version of attR" /label=attR3 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(3963..3982) /label=T7 /note="T7 promoter, forward primer" promoter complement(3964..3982) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3987..4003) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3987..4003) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4105..4124) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 4116..4925 /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(4183..4202) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 5072..5660 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5561..5580 /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 aagttgtaac catggtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac 781 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca 841 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt gcctggtttc 901 cggcaccaga agcggtgccg gaaagctggc tggagtgcga tcttcctgag gccgatactg 961 tcgtcgcccc ctcaaactgg cagatgcacg gttacgatgc gcccatctac tccaacgtga 1021 cctatcccat tacggtcaat ccgccgtttg ttcccacgga gaatccgacg ggttgttact 1081 cgctcacatt taatgttgat gaaagctggc tacaggaagg ccagacgcga attatttttg 1141 atggcgttaa ctcggcgttt catctgtggt gcaacgggcg ctgggtcggt tacggccagg 1201 acagtcgttt gccgtctgaa tttgacctga gcgcattttt acgcgccgga gaaaaccgcc 1261 tcgcggtgat ggtgctgcgc tggagtgacg gcagttatct ggaagatcag gatatgtggc 1321 ggatgagcgg cattttccgt gacgtctcgt tgctgcataa accgactaca caaatcagcg 1381 atttccatgt tgccactcgc tttaatgatg atttcagccg cgctgtactg gaggctgaag 1441 ttcagatgtg cggcgagttg cgtgactacc tacgggtaac agtttcttta tggcagggtg 1501 aaacgcaggt cgccagcggc accgcgcctt tcggcggtga aattatcgat gagcgtggtg 1561 gttatgccga tcgcgtcaca ctacgtctga acgtcgaaaa cccgaaactg tggagcgccg 1621 aaatcccgaa tctctatcgt gcggtggttg aactgcacac cgccgacggc acgctgattg 1681 aagcagaagc ctgcgatgtc ggtttccgcg aggtgcggat tgaaaatggt ctgctgctgc 1741 tgaacggcaa gccgttgctg attcgaggcg ttaaccgtca cgagcatcat cctctgcatg 1801 gtcaggtcat ggatgagcag acgatggtgc aggatatcct gctgatgaag cagaacaact 1861 ttaacgccgt gcgctgttcg cattatccga accatccgct gtggtacacg ctgtgcgacc 1921 gctacggcct gtatgtggtg gatgaagcca atattgaaac ccacggcatg gtgccaatga 1981 atcgtctgac cgatgatccg cgctggctac cggcgatgag cgaacgcgta acgcgaatgg 2041 tgcagcgcga tcgtaatcac ccgagtgtga tcatctggtc gctggggaat gaatcaggcc 2101 acggcgctaa tcacgacgcg ctgtatcgct ggatcaaatc tgtcgatcct tcccgcccgg 2161 tgcagtatga aggcggcgga gccgacacca cggccaccga tattatttgc ccgatgtacg 2221 cgcgcgtgga tgaagaccag cccttcccgg ctgtgccgaa atggtccatc aaaaaatggc 2281 tttcgctacc tggagagacg cgcccgctga tcctttgcga atacgcccac gcgatgggta 2341 acagtcttgg cggtttcgct aaatactggc aggcgtttcg tcagtatccc cgtttacagg 2401 gcggcttcgt ctgggactgg gtggatcagt cgctgattaa atatgatgaa aacggcaacc 2461 cgtggtcggc ttacggcggt gattttggcg atacgccgaa cgatcgccag ttctgtatga 2521 acggtctggt ctttgccgac cgcacgccgc atccagcgct gacggaagca aaacaccagc 2581 agcagttttt ccagttccgt ttatccgggc aaaccatcga agtgaccagc gaatacctgt 2641 tccgtcatag cgataacgag ctcctgcact ggatggtggc gctggatggt aagccgctgg 2701 caagcggtga agtgcctctg gatgtcgctc cacaaggtaa acagttgatt gaactgcctg 2761 aactaccgca gccggagagc gccgggcaac tctggctcac agtacgcgta gtgcaaccga 2821 acgcgaccgc atggtcagaa accgggcaca tcagcgcctg gcagcagtgg cgtctggcgg 2881 aaaacctcag tgtgacgctc cccgccgcgt cccacgccat cccgcatctg accaccagcg 2941 aaatggattt ttgcatcgag ctgggtaata agcgttggca atttaaccgc cagtcaggct 3001 ttctttcaca gatgtggatt ggcgataaaa aacaactgct gacgccgctg cgcgatcagt 3061 tcacccgtgc accgctggat aacgacattg gcgtaagtga agcgacccgc attgacccta 3121 acgcctgggt cgaacgctgg aaggcggcgg gccattacca ggccgaagca gcgtcgttgc 3181 agtgcacggc agatacactt gctgatgcgg cgctgattac gaccgctcac gcgtggcagc 3241 atcaggggaa aaccttattt atcagccgga aaacctaccg gattgatggt agtggtcaaa 3301 tggcgattac cgttgatgtt gaagtggcga gcgatacacc gcatccggcg cggattggcc 3361 tgaactgcca gctggcgcag gtagcagagc gggtaaactg gctcggatta gggccgcaag 3421 aaaactatcc cgaccgccct actgccgcct gttttgaccg ctgggatctg ccattgtcag 3481 acatgtatac cccgtacgtc ttcccgagcg aaaacggtct gcgctgcggg acgcgcgaat 3541 tgaattatgg cccacaccag tggcgcggcg acttccagtt caacatcagc cgctacagtc 3601 aacagcaact gatggaaacc agccatcgcc atctgctgca cgcggaagaa ggcacatggc 3661 tgaatatcga cggtttccat atggggattg gtggcgacga ctcctggagc ccgtcagtat 3721 cggcggaatt ccagctgagc gccggtcgct accattacca gttggtctgg tgtcaaaaat 3781 aagacaactt tgtataataa agttgaacga gaaacgtaaa atgatataaa tatcaatata 3841 ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag 3901 tcactatgaa tcaactactt agatggtatt agtgacctgt actgcagggc ggccgcgata 3961 tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt 4021 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac 4081 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt 4141 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg 4201 ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag 4261 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca 4321 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc 4381 ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc caggtattag 4441 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt 4501 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc 4561 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta 4621 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg 4681 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat 4741 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca 4801 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat 4861 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt 4921 tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg acttgacggg 4981 acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt 5041 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct 5101 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc 5161 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc 5221 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 5281 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 5341 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 5401 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 5461 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 5521 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt 5581 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 5641 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt 5701 gctggccttt tgctcacatg tt //