LOCUS Exported 6135 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Empty LIC vector; adds an mCherry gene to the C-terminus of your protein of interest.. ACCESSION . VERSION . KEYWORDS pcDNA3 mCherry LIC cloning vector (6B) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6135) TITLE 6-series mammalian LIC expression vectors REFERENCE 2 (bases 1 to 6135) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6135 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(163..751) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(243..262) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(922..1782) /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 1545..1564 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1783..1887) /gene="bla" /label=AmpR promoter primer_bind complement(1962..1981) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 2153..2532 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2533..2736 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2687..2707 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2781..2800 /label=T7 /note="T7 promoter, forward primer" promoter 2781..2799 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 2880..3587 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE GRHSTGGMDELYK" primer_bind complement(3022..3040) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 3290..3309 /label=mCherry-F /note="mCherry, forward primer" promoter complement(3606..3624) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(3607..3624) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(3644..3661) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3650..3874 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3920..4348 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4007..4026) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4217..4238 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4357..4377) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4362..4691 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4542..4677 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4604..4623 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4758..5552 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4812..4831) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5422..5441 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5726..5847 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5763..5782) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5817..5836 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5896..5912) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5896..5912) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5909..5931) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5920..5936 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(5944..5974) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5989..6010 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6127..9) /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 61 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 121 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 181 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 241 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 301 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 361 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 421 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 481 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 541 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 601 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 661 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 721 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 781 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 841 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 901 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 961 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 1021 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 1081 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 1141 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 1201 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 1261 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 1321 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 1381 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 1441 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 1501 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 1561 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 1621 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 1681 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 1741 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcatta 1801 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 1861 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga 1921 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc 1981 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg 2041 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt 2101 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga 2161 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg 2221 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc 2281 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat 2341 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat 2401 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat 2461 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc 2521 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac 2581 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa 2641 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt 2701 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact 2761 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc ttggtaccga 2821 gctcggatcc ctacttccaa tccaatattg gtagtgggag caacggcagc agcggatccg 2881 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc 2941 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc 3001 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg 3061 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg 3121 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga 3181 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctc caggacggcg 3241 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc 3301 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc 3361 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg 3421 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca 3481 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg 3541 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtagtaa tctagagggc 3601 cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct 3661 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc 3721 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt 3781 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat 3841 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg 3901 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg 3961 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc 4021 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc 4081 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt 4141 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag 4201 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg 4261 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag 4321 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg 4381 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 4441 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 4501 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc 4561 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg 4621 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag 4681 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag 4741 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt 4801 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg 4861 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg 4921 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg 4981 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg 5041 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca 5101 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc 5161 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc 5221 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca 5281 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga 5341 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg 5401 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg 5461 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg 5521 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga 5581 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag 5641 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct 5701 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata 5761 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg 5821 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag 5881 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc 5941 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta 6001 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca 6061 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc 6121 cgcttcctcg ctcac //