LOCUS Exported 5462 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION pMVP destination vector, empty expression vector backbone w/ Puro selection cassette. ACCESSION . VERSION . KEYWORDS JL302: pMVP/Puro-DEST SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5462) 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 5462) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5462 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 24..128 /gene="bla" /label=AmpR promoter CDS 129..989 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(347..366) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1160..1748 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1649..1668 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1902..1919 /label=L4440 /note="L4440 vector, forward primer" primer_bind 1925..1942 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 1926..1942 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 1966..1982 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind 1966..1982 /label=pBluescriptSK /note="For pBluescript vector" protein_bind 2008..2132 /gene="mutant version of attR" /label=attR1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 2157..2187 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" CDS 2241..2900 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(2307..2326) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" CDS 3242..3547 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 3426..3445 /label=ccdB-fwd /note="ccdB gene, forward primer" protein_bind complement(3588..3712) /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(3727..3743) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3728..3744) /label=pBluescriptKS /note="For pBluescript vector" promoter 3764..4080 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3931..4066 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3993..4012 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 4128..4175 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 4194..4793 /codon_start=1 /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(4833..4852) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4877..4925 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin complement(5101..5459) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5141..5162) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5353..5372 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt 61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa 121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt 181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt 241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt 301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg 361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga 421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa 481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga 541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa 601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca 661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta 721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac 781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc 841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag 901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga 961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt 1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata 1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag 1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc 1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc 1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa 1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg 1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg 1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg 1921 aagctgtaaa acgacggcca gtagctccac cgcggtggcg gccgctctag aactagtgga 1981 tcccccgggc tgcaggaatt cgatatcaca agtttgtaca aaaaagctga acgagaaacg 2041 taaaatgata taaatatcaa tatattaaat tagattttgc ataaaaaaca gactacataa 2101 tactgtaaaa cacaacatat ccagtcacta tggcggccgc attaggcacc ccaggcttta 2161 cactttatgc ttccggctcg tataatgtgt ggattttgag ttaggatccg tcgagatttt 2221 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat 2281 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata 2341 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca 2401 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc 2461 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg 2521 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc 2581 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt 2641 tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca 2701 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg 2761 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg 2821 ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg 2881 agtggcaggg cggggcgtaa acgcgtggat ccggcttact aaaagccaga taacagtatg 2941 cgtatttgcg cgctgatttt tgcggtataa gaatatatac tgatatgtat acccgaagta 3001 tgtcaaaaag aggtatgcta tgaagcagcg tattacagtg acagttgaca gcgacagcta 3061 tcagttgctc aaggcatata tgatgtcaat atctccggtc tggtaagcac aaccatgcag 3121 aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga agggatggct 3181 gaggtcgccc ggtttattga aatgaacggc tcttttgctg acgagaacag gggctggtga 3241 aatgcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt 3301 acagagtgat attattgaca cgcccgggcg acggatggtg atccccctgg ccagtgcacg 3361 tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag 3421 ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt 3481 ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg 3541 aatataaatg tcaggctccc ttatacacag ccagtctgca ggtcgaccat agtgactgga 3601 tatgttgtgt tttacagtat tatgtagtct gttttttatg caaaatctaa tttaatatat 3661 tgatatttat atcattttac gtttctcgtt cagctttctt gtacaaagtg gtgatatcaa 3721 gcttatcgat accgtcgacc tcgacatggt catagctgtt gacggtgtgg aaagtcccca 3781 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt 3841 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 3901 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 3961 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct 4021 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa 4081 aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat 4141 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatgaccg 4201 agtacaagcc taccgtgcgc ctggccactc gcgatgatgt gccccgcgcc gtccgcactc 4261 tggccgccgc tttcgccgac taccccgcta cccggcacac cgtggacccc gaccggcaca 4321 tcgagcgtgt gacagagttg caggagctgt tcctgacccg cgtcgggctg gacatcggca 4381 aggtgtgggt agccgacgac ggcgcggccg tggccgtgtg gactaccccc gagagcgttg 4441 aggccggcgc cgtgttcgcc gagatcggcc cccgaatggc cgagctgagc ggcagccgcc 4501 tggccgccca gcagcaaatg gagggcctgc ttgcccccca tcgtcccaag gagcctgcct 4561 ggtttctggc cactgtagga gtgagccccg accaccaggg caagggcttg ggcagcgccg 4621 tcgtgttgcc cggcgtagag gccgccgaac gcgccggtgt gcccgccttt ctcgaaacaa 4681 gcgcaccaag aaaccttcca ttctacgagc gcctgggctt caccgtgacc gccgatgtcg 4741 aggtgcccga gggacctagg acctggtgta tgacacgaaa acctggcgcc taacgtaccg 4801 gttagtaatg agtttaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa 4861 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt 4921 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc 4981 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg 5041 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatccggt 5101 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc 5161 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg 5221 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact 5281 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt 5341 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc 5401 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc 5461 ag //