LOCUS Exported 6236 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of SNAP-tagged Histone H2B in mammalian cells. ACCESSION . VERSION . KEYWORDS pSNAPf-H2B Control Plasmid SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6236) AUTHORS Ana Egana TITLE Egana Lab NEB plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 6236) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6236 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(157..174) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 384..403 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 599..1402 /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(662..681) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 1272..1291 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 1532..1653 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1569..1588) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(1755..1771) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1755..1771) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1768..1790) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1779..1795 /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(1803..1833) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1848..1869 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1986..2003) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2157..2745) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2237..2256) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2916..3776) /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 3539..3558 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3777..3881) /gene="bla" /label=AmpR promoter primer_bind complement(3956..3975) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 4147..4526 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4527..4730 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4681..4701 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 4775..4794 /label=T7 /note="T7 promoter, forward primer" promoter 4775..4793 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 4836..5213 /codon_start=1 /gene="HIST1H2BJ" /product="human histone H2B" /label=H2B /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP GELAKHAVSEGTKAITKYTSAK" CDS 5268..5813 /codon_start=1 /product="human O6-alkylguanine-DNA-alkyltransferase variant that forms covalent bonds with benzylguanine derivatives" /label=SNAP-tag(R) /note="fast-reacting variant, optimized for mammalian expression" /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA VKEWLLAHEGHRLGKPGLG" intron 5924..6153 /label=chimeric intron /note="chimera between introns from adenovirus and immunoglobulin heavy chain genes" primer_bind 6136..6155 /label=pMT2-F /note="Synthetic intron, forward primer" misc_feature 6226..563 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" ORIGIN 1 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct 61 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc 121 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc 181 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg 241 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa 301 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt 361 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg 421 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta 481 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt 541 tcctttgaaa aacacgatga taagcttgcc acaacccggg ataattcctg cagccaatat 601 gggatcggcc attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag 661 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg 721 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa 781 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc 841 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc 901 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga 961 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa 1021 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct 1081 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat 1141 gcccgacggc gatgatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt 1201 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta 1261 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga 1321 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg 1381 ccttcttgac gagttcttct gaggggatca attctctaga taactgatca taatcagcca 1441 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct 1501 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta 1561 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 1621 ttgtggtttg tccaaactca tcaatgtatc ttaacgcgtc gagtgcattc tagttgtggt 1681 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc 1741 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca 1801 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa 1861 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 1921 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 1981 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 2041 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 2101 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 2161 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 2221 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 2281 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 2341 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 2401 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 2461 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 2521 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 2581 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc 2641 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 2701 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 2761 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 2821 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 2881 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 2941 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 3001 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 3061 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 3121 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 3181 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 3241 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 3301 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 3361 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 3421 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 3481 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 3541 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 3601 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 3661 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 3721 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 3781 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 3841 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 3901 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa 3961 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg 4021 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca 4081 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata 4141 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc 4201 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac 4261 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa 4321 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag 4381 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc 4441 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct 4501 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg 4561 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt 4621 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga 4681 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa 4741 ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc 4801 aagcttggta ccgagctcgg atcgatatcg aattgatgcc agagccagcg aagtctgctc 4861 ccgccccgaa aaagggctcc aagaaggcgg tgactaaggc gcagaagaaa ggcggcaaga 4921 agcgcaagcg cagccgcaag gagagctatt ccatctatgt gtacaaggtt ctgaagcagg 4981 tccaccctga caccggcatt tcgtccaagg ccatgggcat catgaattcg tttgtgaacg 5041 acattttcga gcgcatcgca ggtgaggctt cccgcctggc gcattacaac aagcgctcga 5101 ccatcacctc cagggagatc cagacggccg tgcgcctgct gctgcctggg gagttggcca 5161 agcacgccgt gtccgagggt actaaggcca tcaccaagta caccagcgct aaggctagcg 5221 atatcggcgc gccagcattt aaatctgtac agaccggtga attcaccatg gacaaagact 5281 gcgaaatgaa gcgcaccacc ctggatagcc ctctgggcaa gctggaactg tctgggtgcg 5341 aacagggcct gcaccgtatc atcttcctgg gcaaaggaac atctgccgcc gacgccgtgg 5401 aagtgcctgc cccagccgcc gtgctgggcg gaccagagcc actgatgcag gccaccgcct 5461 ggctcaacgc ctactttcac cagcctgagg ccatcgagga gttccctgtg ccagccctgc 5521 accacccagt gttccagcag gagagcttta cccgccaggt gctgtggaaa ctgctgaaag 5581 tggtgaagtt cggagaggtc atcagctaca gccacctggc cgccctggcc ggcaatcccg 5641 ccgccaccgc cgccgtgaaa accgccctga gcggaaatcc cgtgcccatt ctgatcccct 5701 gccaccgggt ggtgcagggc gacctggacg tggggggcta cgagggcggg ctcgccgtga 5761 aagagtggct gctggcccac gagggccaca gactgggcaa gcctgggctg ggtcctgcag 5821 gcggatccgc gtttaaactc gaggttaatt aatgagcggc cgcatagata actgatccag 5881 tgtgctggaa ttaattcgct gtctgcgagg gccagctgtt ggggtgagta ctccctctca 5941 aaagcgggca tgacttctgc gctaagattg tcagtttcca aaaacgagga ggatttgata 6001 ttcacctggc ccgcggtgat gcctttgagg gtggccgcgt ccatctggtc agaaaagaca 6061 atctttttgt tgtcaagctt gaggtgtggc aggcttgaga tctggccata cacttgagtg 6121 acaatgacat ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcagg 6181 tcgagcatgc atctagggcg gccaattccg cccctctccc ccccccccct ctccct //