LOCUS Exported 7238 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION For mRNA synthesis or expression in mammalian cells. ACCESSION . VERSION . KEYWORDS pcDNA-flag-mKdm4d-polyA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7238) AUTHORS Matoba S, Liu Y, Lu F, Iwabuchi KA, Shen L, Inoue A, Zhang Y TITLE Embryonic Development following Somatic Cell Nuclear Transfer Impeded by Persisting Histone Methylation. JOURNAL Cell. 2014 Nov 6;159(4):884-95. doi: 10.1016/j.cell.2014.09.055. Epub 2014 Oct 30. PUBMED 25417163 REFERENCE 2 (bases 1 to 7238) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7238 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 911..934 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 2741..2770 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2786..2803 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(2826..2843) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2832..3056 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3102..3530 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3189..3208) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3399..3420 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3539..3559) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3544..3874 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3725..3860 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3787..3806 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3941..4735 /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(3995..4014) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4605..4624 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4911..5032 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4948..4967) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5002..5021 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5081..5097) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5081..5097) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5094..5116) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5105..5121 /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(5129..5159) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5174..5195 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5312..5329) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5483..6071) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5563..5582) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6242..7102) /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 6865..6884 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7103..7207) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt 901 taagcttatg gactacaaag acgatgacga caagggatcc actagtaacg gccgccagtg 961 tgctggaatt cacaagacga agtccacatg tgctcagaat ccaaattgca gcataatgat 1021 atttcgtcca accaaagaag agtttaatga ttttgacaaa tacattgctt acatggagtc 1081 ccaaggggca caccgagctg gactggccaa ggtcatccca ccaaaagaat ggagggccag 1141 gcagtcttat gacaatatca gcaacatctt aatagcaact cccctgcagc aagtggtctc 1201 tgggcaggca ggcgtgttca cccaatacca taagaagaag aaaggcatga cagtggggga 1261 gtaccgtgag ctggccaaca gcaaaaagta ccagaccccg ccacacctgg attttgaaga 1321 tttggagcga aaatactgga agaatcgcct gtatgagtca ccgatttatg gtgctgacgt 1381 cagcggctcc ctgtttgatg ggaagactca acagtggaat gtgggtcacc tgggaacaat 1441 tcaagaccta ttggaacagg aatgtggcat agtgattgag ggcgtcaaca cgccctacct 1501 gtactttggc atgtggaaga ccacctttgc gtggcacacg gaggacatgg acctgtacag 1561 tatcaactac ctacactttg ggcagcccaa gacctggtat gctgtgcccc ctgagcatgg 1621 caggcgcctg gagcgcctgg ccagggaact cttccctggc agctcccagg gctgccaggc 1681 cttcctgagg cacaaggtgg cgctcatctc gcccactgtg cttaaggaga atggcatccc 1741 ctttggtcgc atcacccagg aggctgggga gttcatggtc acctttccct atggctacca 1801 cgcgggcttc aaccatggct tcaactgcgc agaggccatc aattttgcca ccccaaggtg 1861 gattgactat ggcaaggtgg catctcagtg cagctgtggg gaggccaggg tgagcttctc 1921 catggacgcc tttgtgagga tcttgcagcc tgagcgatat gaactgtgga aacgtggcca 1981 agatcaggca gttgtggacc acacagagac tatggtgtct accagtcagg agctcaccac 2041 ccggcgggtg accaaagcac caagaaaaac ttggggcttg aagcgtctcc ggcttcgcca 2101 ggtctcacga tctcttctgc ctatagccac ggtaagtaac gttccttgca acatgcaggt 2161 gtgccacacc tccaggcaac catcagatgt gaaaggtgat gatgtccaga agtctgactc 2221 agccagagcc tcaccacatc ctctgagtct gccttcttct ggtcacatgt caactagaag 2281 atgtagtctt ggtcgtcgtc cttgtgaact aggagctcag gagtcctcca atggagctcc 2341 agtcaagagg caacttccag caggcagaga tgacacaagc cccagtccag agcttcagcc 2401 ccaggctgtg agtggagact taatagtcga ctcaggactt gtgaaccctg gcccacagca 2461 tcttatgaca gcttctgaag ggggattgac ctccgacccc taagaattct gcagatatcc 2521 atcacactgg cggccgctcg actatgggca ccaaagaacc tgtaaacgtt atctttttaa 2581 attgaatgtg cacaaataaa agtttggaaa agaaaaaaaa aaaaaaaaaa aaaaaaaaaa 2641 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaatacgc 2701 atcatacgct gtggcgtacc tcgagtctag agggcccttc gaacaaaaac tcatctcaga 2761 agaggatctg aatatgcata ccggtcatca tcaccatcac cattgagttt aaacccgctg 2821 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc 2881 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc 2941 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa 3001 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc 3061 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc 3121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct 3181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg 3241 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga 3301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt 3361 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg 3421 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc 3481 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg 3541 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc caggcaggca gaagtatgca 3601 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg 3661 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc 3721 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat 3781 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg 3841 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat 3901 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt 3961 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca 4021 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct 4081 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct 4141 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc 4201 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct 4261 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga 4321 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg 4381 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc 4441 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac 4501 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat 4561 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga 4621 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc 4681 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg 4741 actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 4801 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 4861 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta 4921 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 4981 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 5041 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt 5101 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 5161 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 5221 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 5281 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 5341 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 5401 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 5461 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 5521 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 5581 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 5641 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca 5701 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 5761 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 5821 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 5881 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct 5941 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 6001 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 6061 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 6121 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 6181 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 6241 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 6301 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 6361 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 6421 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 6481 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 6541 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 6601 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 6661 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 6721 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 6781 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 6841 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 6901 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 6961 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 7021 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 7081 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 7141 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 7201 ttccgcgcac atttccccga aaagtgccac ctgacgtc //