LOCUS Exported 7359 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses Mouse Nanog K137A Mutants. It is modified from pPyCAG-Nanog-IP (Addgene Plasmid #13838).. ACCESSION . VERSION . KEYWORDS pPyCAG-Nanog(K137A)-IP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7359) TITLE Plasmids of NANOG HD Mutants JOURNAL Unpublished REFERENCE 2 (bases 1 to 7359) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7359 /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..4018 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(3599..3616) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 3826..3845 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 4019..4618 /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(4019..4038) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 4515..4535 /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind complement(4699..4716) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4705..4929 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(4976..4992) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4976..4992) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4989..5011) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5000..5016 /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(5024..5054) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5069..5090 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5149..5344 /label=SV40 promoter /note="SV40 early promoter" rep_origin 5195..5330 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5257..5276 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 5350..5484 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5552..5569) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5723..6311) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5803..5822) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6482..7342) /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 7105..7124 /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 gatgcaagaa ctctcctcca ttctgaacct gagctatgcg 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 cctaacgtta ctggccgaag ccgcttggaa 3481 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat 3541 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct 3601 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct 3661 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc 3721 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa 3781 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc 3841 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg 3901 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc 3961 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat 4021 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg 4081 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg 4141 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat 4201 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag 4261 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc 4321 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc 4381 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag 4441 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga 4501 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga 4561 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg 4621 cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcgatgat 4681 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc 4741 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 4801 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 4861 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg 4921 gctctatggc ttctgaggcg gaaagaacct gcagcccaag cttggcgtaa tcatggtcat 4981 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa 5041 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc 5101 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta 5161 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc 5221 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc 5281 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg 5341 caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 5401 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 5461 aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag 5521 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 5581 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 5641 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 5701 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 5761 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 5821 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 5881 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 5941 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 6001 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 6061 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 6121 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 6181 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 6241 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 6301 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 6361 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 6421 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 6481 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 6541 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 6601 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 6661 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 6721 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 6781 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 6841 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 6901 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 6961 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 7021 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 7081 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 7141 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 7201 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 7261 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 7321 cacggaaatg ttgaatactc atactcttcc tttttcaat //