LOCUS Exported 3545 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pMVP L3-L2 entry plasmid, contains eCFP + polyA for 3- or 4-component MultiSite Gateway Pro assembly. Allows C-term eCFP fusion to gene of interest.. ACCESSION . VERSION . KEYWORDS LC701: pMVP (L3-L2) eCFP + polyA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3545) 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 3545) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3545 /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..1399 /codon_start=1 /product="enhanced CFP" /label=ECFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(725..746) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(986..1005) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1333..1354 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1435..1452) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1441..1648 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" protein_bind complement(1657..1756) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1786..1805) /label=T7 /note="T7 promoter, forward primer" promoter complement(1787..1805) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1810..1826) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1810..1826) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1928..1947) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1939..2748 /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(2006..2025) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2895..3483 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3384..3403 /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 ggcggatcca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc 721 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg 781 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct 841 gcccgtgccc tggcccaccc tcgtgaccac cctgacctgg ggcgtgcagt gcttcagccg 901 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt 961 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa 1021 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga 1081 cggcaacatc ctggggcaca agctggagta caactacatc agccacaacg tctatatcac 1141 cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga 1201 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt 1261 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga 1321 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat 1381 ggacgagctg tacaagtaag aattctgaag cttagcctag agctcgctga tcagcctcga 1441 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc 1501 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc 1561 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt 1621 gggaagagaa tagcaggcat gctggggagg tacctgaccc agctttcttg tacaaagttg 1681 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa 1741 ataaaatcat tatttgccat ccagctgcag ggcggccgcg atatccccta tagtgagtcg 1801 tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt 1861 tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac 1921 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat 1981 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca 2041 ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat 2101 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg 2161 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta 2221 ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca 2281 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt 2341 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg 2401 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa 2461 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat 2521 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat 2581 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc 2641 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct 2701 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt 2761 aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc aagctcatga 2821 ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag 2881 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa 2941 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg 3001 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag 3061 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg 3121 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga 3181 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc 3241 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc 3301 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga 3361 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt 3421 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg 3481 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac 3541 atgtt //