LOCUS Exported 7361 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses Mouse Nanog M125A Mutants. It is modified from pPyCAG-Nanog-IP (Addgene Plasmid #13838).. ACCESSION . VERSION . KEYWORDS pPyCAG-Nanog(M125A)-IP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7361) TITLE Plasmids of NANOG HD Mutants JOURNAL Unpublished REFERENCE 2 (bases 1 to 7361) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7361 /organism="synthetic DNA construct" /mol_type="other DNA" polyA_signal 576..710 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" enhancer 724..1103 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1105..1382 /label=chicken beta-actin promoter intron 1383..2399 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 2323..2345 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 2407..2426 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" protein_bind 2437..2461 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(3390..3414) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" misc_feature 3432..4020 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(3601..3618) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 3828..3847 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 4021..4620 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(4021..4040) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 4517..4537 /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind complement(4701..4718) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4707..4931 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(4978..4994) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4978..4994) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4991..5013) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5002..5018 /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(5026..5056) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5071..5092 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5151..5346 /label=SV40 promoter /note="SV40 early promoter" rep_origin 5197..5332 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5259..5278 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 5352..5486 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5554..5571) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5725..6313) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5805..5824) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6484..7344) /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 7107..7126 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 gaccgctttg gaaaaacaaa gactgtattt cctggaaatt aatgtttatt caataaactg 61 tgtattcagc tatattcaca tagtggtgag gctgaaatga ggcgggaaga ggcggttggg 121 gcttaattat atcaatttgg gtggccccac agcgcctcca aggcgccagt cctgttttga 181 caagttgcct ctggaagcct ctcttctttt tctccagagt aagcggaggc caggggcccc 241 cggcctctgc ttaatactaa aaaaaacagc tgttgtcata gtaatgattg ggtggaaaca 301 ttccaggcct gggtggagag gctttttgct tcctcttgca aaaccacact gacattccag 361 gcctgggtgg agaggctttt tgcttcctct tgcaaaacca cactgccctc tggagggcag 421 ttgcctagca actaattaaa agaggatgtc gcacggccag ctgcggtcag ttagtcactt 481 cctgcttaac tgacttgaca ttttctattt taagagtcgg gaggaaaatt actgtgttgg 541 aggccctccg ccatcttctg aagctgaatc gaattaactt gtttattgca gcttataatg 601 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt 661 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc tgatatcatc 721 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 781 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 841 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 901 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 961 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 1021 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 1081 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca 1141 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag 1201 cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc 1261 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt 1321 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg 1381 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc 1441 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc 1501 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg 1561 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt 1621 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc 1681 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg 1741 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg 1801 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc 1861 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc 1921 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg 1981 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg 2041 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg 2101 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca 2161 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg 2221 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc 2281 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg 2341 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcntacagc 2401 tcctgggcaa cgtgctggtt attgtgctga attatcacaa gtttgtacaa aaaagcaggc 2461 tctgacatga gtgtgggtct tcctggtccc cacagtttgc ctagttctga ggaagcatcg 2521 aattctggga acgcctcatc aatgcctgca gtttttcatc ccgagaacta ttcttgctta 2581 caagggtctg ctactgagat gctctgcaca gaggctgcct ctcctcgccc ttcctctgaa 2641 gacctgcctc ttcaaggcag ccctgattct tctaccagtc ccaaacaaaa gctctcaagt 2701 cctgaggctg acaagggccc tgaggaggag gagaacaagg tccttgccag gaagcagaag 2761 atgcggactg tgttctctca ggcccagctg tgtgcactca aggacaggtt tcagaagcag 2821 aagtacctca gcctccagca ggcgcaagaa ctctcctcca ttctgaacct gagctataag 2881 caggttaaga cctggtttca aaaccaaagg atgaagtgca agcggtggca gaaaaaccag 2941 tggttgaaga ctagcaatgg tctgattcag aagggctcag caccagtgga gtatcccagc 3001 atccattgca gctatcccca gggctatctg gtgaacgcat ctggaagcct ttccatgtgg 3061 ggcagccaga cttggaccaa cccaacttgg agcagccaga cctggaccaa cccaacttgg 3121 aacaaccaga cctggaccaa cccaacttgg agcagccagg cctggaccgc tcagtcctgg 3181 aacggccagc cttggaatgc tgctccgctc cataacttcg gggaggactt tctgcagcct 3241 tacgtacagt tgcagcaaaa cttctctgcc agtgatttgg aggtgaattt ggaagccact 3301 agggaaagcc atgcgcattt tagcacccca caagccttgg aattattcct gaactactct 3361 gtgactccac caggtgaaat atgagactta cccagctttc ttgtacaaag tggtgataat 3421 tccgcccccc ccccctctcc ctcccccccc cccctaacgt tactggccga agccgcttgg 3481 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca 3541 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc 3601 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag 3661 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg 3721 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac 3781 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa 3841 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc 3901 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc 3961 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc 4021 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta 4081 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac 4141 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac 4201 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag 4261 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt 4321 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag 4381 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc 4441 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg 4501 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc 4561 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga 4621 cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg cacgacccca tgcatcgatg 4681 atctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg 4741 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 4801 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 4861 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt 4921 gggctctatg gcttctgagg cggaaagaac ctgcagccca agcttggcgt aatcatggtc 4981 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 5041 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 5101 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat 5161 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 5221 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 5281 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt 5341 tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 5401 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 5461 tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag 5521 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 5581 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 5641 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 5701 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 5761 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 5821 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 5881 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 5941 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 6001 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 6061 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 6121 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 6181 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 6241 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa 6301 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga 6361 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct 6421 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga 6481 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 6541 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg 6601 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat 6661 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat 6721 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 6781 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 6841 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa 6901 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc 6961 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt 7021 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 7081 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt 7141 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag 7201 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 7261 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 7321 gacacggaaa tgttgaatac tcatactctt cctttttcaa t //