LOCUS Exported 10381 bp ds-DNA circular SYN 01-AUG-2016 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10381) AUTHORS . TITLE Direct Submission JOURNAL Exported 2015��10��12�� from SnapGene Viewer 2.8.2 http://www.snapgene.com REFERENCE 2 (bases 1 to 10381) AUTHORS . TITLE Direct Submission JOURNAL Exported May 25, 2018 from SnapGene Viewer 4.1.9 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10381 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 239..1419 /label=EF-1-alpha promoter /note="EF-1-alpha promoter;strong constitutive promoter for human elongation factor EF-1-alpha" intron 469..1411 /label=EF-1-alpha intron A /note="EF-1-alpha intron A;intron upstream of the start codon of human EF-1-alpha" promoter 1469..1487 /label=T7 promoter /note="T7 promoter;promoter for bacteriophage T7 RNA polymerase" CDS 1516..1536 /codon_start=1 /product="nuclear localization signal of SV40 large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1555..2163 /codon_start=1 /label=DNMT3L /translation="MFETVPVWRRQPVRVLSLFEDIKKELTSLGFLESGSDPGQLKHVV DVTDTVRKDVEEWGPFDLVYGATPPLGHTCDRPPSWYLFQFHRLLQYARPKPGSPRPFF WMFVDNLVLNKEDLDVASRFLEMEPVTIPDVHGGSLQNAVRVWSNIPAIRSSRHWALVS EEELSLLAQNKQSSKLAAKWPTKLVKNCFLPLREYFKYFS" misc_feature 2164..2211 /label=linker1 CDS 2212..3069 /codon_start=1 /label=DNMT3A /translation="PAEKRKPIRVLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSIT VGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRL FFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAH RARYFWGNLPGMNRPLASTVNDKLELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHF PVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKE YFACV" misc_feature 3070..3116 /label=linker2 misc_feature 3142..3144 /label=D10A misc_feature 5632..5634 /label=H840A misc_feature 5701..5703 /label=H863A CDS 7225..7245 /codon_start=1 /product="nuclear localization signal of SV40 large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 7276..7332 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 7333..8043 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTRGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" polyA_signal 8069..8293 /label=bGH poly /note="bGH poly(A) signal;bovine growth hormone polyadenylation signal" rep_origin complement(8626..9214) /direction=LEFT /label=ori /note="ori;high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(9385..10245) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=bla /note="AmpR;confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(10246..10350) /gene="bla" /label=bla AmpR promoter /note="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 cgtgtgaggc 241 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga 301 ggggtcggca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat 361 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta 421 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg 481 tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact 541 tccacgcccc tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg 601 ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg 661 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc 721 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg 781 gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc 841 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg 901 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc 961 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc 1021 agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag 1081 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc 1141 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga 1201 ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat 1261 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta 1321 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac 1381 agtggttcaa agtttttttc ttccatttca ggtgtcgtgg agctctctgg ctaactagag 1441 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg 1501 gctagcacca tgggacctaa gaaaaagagg aaggtggcgg ccgctggcgg cagcatgttc 1561 gaaaccgtgc ctgtgtggag gagacagcca gtccgggtgc tgtccctttt tgaagacatc 1621 aagaaagagc tgacgagttt gggctttttg gaaagtggtt ctgacccggg acaactgaag 1681 catgtggttg atgtcacaga cacagtgagg aaggatgtgg aggagtgggg acccttcgat 1741 cttgtgtacg gcgccacacc tcccctgggc cacacctgtg accgtcctcc cagctggtac 1801 ctgttccagt tccaccggct cctgcagtac gcacggccca agccaggcag ccccaggccc 1861 ttcttctgga tgttcgtgga caatctggtg ctgaacaagg aagacctgga cgtcgcatct 1921 cgcttcctgg agatggagcc agtcaccatc ccagatgtcc acggcggatc cttgcagaat 1981 gctgtccgcg tgtggagcaa catcccagcc ataaggagca gcaggcactg ggctctggtt 2041 tcggaagaag aattgtccct gctggcccag aacaagcaga gctcgaagct cgcggccaag 2101 tggcccacca agctggtgaa gaactgcttt ctccccctaa gagaatattt caagtatttt 2161 tcaagcagcg ggcgcagctt cagcagcgga ctagtgcccc gcggcagcca cccagctgag 2221 aagaggaagc ccatccgggt gctgtctctc tttgatggaa tcgctacagg gctcctggtg 2281 ctgaaggact tgggcattca ggtggaccgc tacattgcct cggaggtgtg tgaggactcc 2341 atcacggtgg gcatggtgcg gcaccagggg aagatcatgt acgtcgggga cgtccgcagc 2401 gtcacacaga agcatatcca ggagtggggc ccattcgatc tggtgattgg gggcagtccc 2461 tgcaatgacc tctccatcgt caaccctgct cgcaagggcc tctacgaggg cactggccgg 2521 ctcttctttg agttctaccg cctcctgcat gatgcgcggc ccaaggaggg agatgatcgc 2581 cccttcttct ggctctttga gaatgtggtg gccatgggcg ttagtgacaa gagggacatc 2641 tcgcgatttc tcgagtccaa ccctgtgatg attgatgcca aagaagtgtc agctgcacac 2701 agggcccgct acttctgggg taaccttccc ggtatgaaca ggccgttggc atccactgtg 2761 aatgataagc tggagctgca ggagtgtctg gagcatggca ggatagccaa gttcagcaaa 2821 gtgaggacca ttactacgag gtcaaactcc ataaagcagg gcaaagacca gcattttcct 2881 gtcttcatga atgagaaaga ggacatctta tggtgcactg aaatggaaag ggtatttggt 2941 ttcccagtcc actatactga cgtctccaac atgagccgct tggcgaggca gagactgctg 3001 ggccggtcat ggagcgtgcc agtcatccgc cacctcttcg ctccgctgaa ggagtatttt 3061 gcgtgtgtga gcggcagcga gacccccggt accagcgaga gcgccacccc cgagagcgac 3121 aagaaatact ctattggact ggctatcggg acaaactccg ttggctgggc cgtcataacc 3181 gacgagtata aggtgccaag caagaaattc aaggtgctgg gtaatactga ccgccattca 3241 atcaagaaga acctgatcgg agcactcctc ttcgactccg gtgaaaccgc tgaagctact 3301 cggctgaagc ggaccgcaag gcggagatac acccgccgca agaatcggat atgttatctg 3361 caagagatct ttagcaacga aatggctaag gtggacgact ccttctttca ccgcctggaa 3421 gagagctttc tggtggagga ggataagaaa cacgagaggc accctatatt cggaaatatc 3481 gtggatgagg tggcttacca tgaaaagtat cctacaatct accatctgag gaagaagctg 3541 gtggacagca ccgataaagc agacctgagg ctcatctatc tggccctggc tcatatgata 3601 aagtttagag gacactttct gatcgagggc gacctgaatc ccgataattc cgatgtggat 3661 aaactcttca ttcaactggt gcagacatat aaccaactgt tcgaggagaa tcccataaac 3721 gcttctggtg tggatgccaa ggctattctg tccgctcggc tgtccaagtc acgcagactg 3781 gagaatctga ttgcccaact gccaggagaa aagaagaacg gcctgtttgg gaacctcatc 3841 gccctgagcc tgggcctgac acctaacttc aagtccaatt ttgatctggc cgaagatgct 3901 aaactccagc tctccaagga cacctatgac gatgatctgg acaacctgct cgcacagata 3961 ggcgaccagt acgccgatct ctttctggct gctaagaatc tctccgacgc cattctgctg 4021 agcgacatac tccgggtcaa cactgagatc accaaagcac ctctgagcgc ctccatgata 4081 aaacgctatg atgaacacca tcaagacctg actctgctca aagccctcgt gaggcaacag 4141 ctgccagaga agtacaaaga gatattcttc gaccagagca agaatggata tgccggatac 4201 atcgatggcg gagcatcaca ggaagaattt tacaagttca tcaaaccaat cctcgagaag 4261 atggacggta ctgaagagct gctggtgaag ctgaacaggg aggacctgct gaggaagcag 4321 aggacctttg ataatggctc cattccacat cagatacacc tgggagagct gcatgcaatc 4381 ctccgcaggc aggaggattt ctatcctttc ctgaaggata accgggagaa gatagagaag 4441 atcctgacct tcaggatccc ttattacgtc ggccctctgg ctagaggcaa ctcccgcttc 4501 gcttggatga ccaggaaatc tgaggagaca attactcctt ggaacttcga agaggtcgtg 4561 gataagggcg caagcgccca gtcattcatc gaacggatga ccaatttcga taagaacctg 4621 ccaaacgaga aggtcctgcc caaacattca ctcctgtacg agtatttcac cgtctataac 4681 gagctgacta aagtgaagta cgtgaccgag ggcatgagga agcctgcctt cctgtccgga 4741 gagcagaaga aggctatcgt tgatctgctc ttcaagacta atagaaaggt gacagtgaag 4801 cagctcaagg aggattactt taagaagatc gaatgctttg actcagtgga aatctctggc 4861 gtggaggacc gctttaatgc cagcctgggc acttaccatg atctgctgaa gataatcaaa 4921 gacaaagatt tcctcgataa tgaggagaac gaggacatcc tggaagatat cgtgctgacc 4981 ctgactctgt tcgaggatag agagatgatc gaagagcgcc tgaagaccta tgcccatctg 5041 tttgacgata aagtcatgaa acagctcaag cggcggcgct acactgggtg gggtagactc 5101 tccaggaaac tcataaacgg catccgcgac aaacagagcg gaaagaccat cctggatttc 5161 ctgaaatccg acggattcgc taacaggaac ttcatgcaac tgattcacga tgactctctg 5221 acatttaaag aggacatcca gaaggcacag gtgagcggtc aaggcgacag cctgcacgag 5281 cacatcgcca acctcgctgg atcacccgcc ataaagaagg gaatactgca gacagtcaag 5341 gtcgtggacg aactcgtcaa agtgatgggt cggcacaagc cagagaatat cgttatcgaa 5401 atggcaaggg agaaccaaac cacccagaag ggccagaaga actctcggga acggatgaaa 5461 agaatcgaag agggaattaa ggagctggga tctcagatac tgaaggagca ccctgtggag 5521 aatacacagc tccagaacga gaaactctac ctgtactacc tccagaacgg gcgggacatg 5581 tacgttgacc aggaactcga catcaaccgg ctgtccgatt atgacgtgga cgctattgtt 5641 ccacagtcct tcctcaaaga tgactccatt gacaacaagg tgctgaccag atccgataag 5701 gcccgcggta agtctgacaa tgttccatca gaagaggtgg tcaagaagat gaagaattac 5761 tggcggcagc tcctcaacgc caaactgatc acccagcgga agtttgacaa tctgactaag 5821 gcagaaagag gaggtctgag cgaactcgac aaggccggct ttattaagag gcaactggtc 5881 gaaacacgcc agattaccaa acacgtggca caaatcctcg actctaggat gaacactaag 5941 tacgatgaga acgataagct gatcagggaa gtgaaagtga taactctgaa gagcaagctg 6001 gtgtctgact tccggaagga ctttcaattc tacaaagttc gcgaaataaa caattaccat 6061 catgctcacg atgcctatct caatgctgtc gttggcaccg ccctgatcaa gaaataccct 6121 aaactggagt ctgagttcgt gtacggtgac tataaagtct acgatgtgag gaagatgata 6181 gcaaagtctg agcaagagat tggcaaagcc accgccaagt acttcttcta ctctaatatc 6241 atgaatttct ttaagactga gataaccctg gctaacggcg aaatccggaa gcgcccactg 6301 atcgaaacaa acggagaaac aggagaaatc gtgtgggata aaggcaggga cttcgcaact 6361 gtgcggaagg tgctgtccat gccacaagtc aatatcgtga agaagaccga agtgcagacc 6421 ggcggattct caaaggagag catcctgcca aagcggaact ctgacaagct gatcgccagg 6481 aagaaagatt gggacccaaa gaagtatggc ggtttcgatt cccctacagt ggcttattcc 6541 gttctggtcg tggcaaaagt ggagaaaggc aagtccaaga aactcaagtc tgttaaggag 6601 ctgctcggaa ttactattat ggagagatcc agcttcgaga agaatccaat cgatttcctg 6661 gaagctaagg gctataaaga agtgaagaaa gatctcatca tcaaactgcc caagtactct 6721 ctctttgagc tggagaatgg taggaagcgg atgctggcct ccgccggaga gctgcagaaa 6781 ggaaacgagc tggctctgcc ctccaaatac gtgaacttcc tgtatctggc ctcccactac 6841 gagaaactca aaggtagccc tgaagacaat gagcagaagc aactctttgt tgagcaacat 6901 aaacactacc tggacgaaat cattgaacag attagcgagt tcagcaagcg ggttattctg 6961 gccgatgcaa acctcgataa agtgctgagc gcatataata agcacaggga caagccaatt 7021 cgcgaacaag cagagaatat tatccacctc tttactctga ctaatctggg cgctcctgct 7081 gccttcaagt atttcgatac aactattgac aggaagcggt acacctctac caaagaagtt 7141 ctcgatgcca ccctgataca ccagtcaatt accggactgt acgagactcg catcgacctg 7201 tctcagctcg gcggcgacgg ttctcccaag aagaagagga aagtctcgag cggtggagct 7261 gcaggaggat ccggcgccac gaacttctct ctgttaaagc aagcaggaga cgtggaagaa 7321 aaccccggtc ctatggtgag caagggcgag gaggataaca tggccatcat caaggagttc 7381 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc 7441 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaggggt 7501 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc 7561 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc 7621 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc 7681 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc 7741 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac 7801 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc 7861 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc 7921 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg 7981 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag 8041 taagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt 8101 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa 8161 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg 8221 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg 8281 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccat 8341 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 8401 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 8461 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 8521 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 8581 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 8641 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 8701 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 8761 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 8821 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 8881 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 8941 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 9001 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 9061 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 9121 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 9181 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 9241 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 9301 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 9361 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 9421 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 9481 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 9541 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 9601 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 9661 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 9721 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 9781 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 9841 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 9901 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 9961 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 10021 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 10081 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 10141 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 10201 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 10261 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 10321 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 10381 c //