LOCUS Exported 7272 bp ds-DNA circular SYN 14-JUL-2021 DEFINITION Expresses SOX2 in mammalian cells. ACCESSION . VERSION . KEYWORDS MAC_N_SOX2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7272) TITLE MAC-N tagged TF-plasmids REFERENCE 2 (bases 1 to 7272) AUTHORS . TITLE Direct Submission JOURNAL Exported Jul 14, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7272 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 586..604 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 607..625 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 635..659 /label=LNCX /note="Human CMV promoter, forward primer" CDS 827..1789 /codon_start=1 /product="promiscuous R118G mutant of the E. coli biotin protein ligase (Cronan, 2005)" /label=BirA(R118G) /note="also known as BirA* or BioID" /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG DACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK" CDS 1790..1816 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1847..1870 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 1907..1930 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" protein_bind 1943..1967 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(2928..2952) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind complement(2990..3007) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2996..3220 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(3353..3372) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 3504..3551 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS 3559..4581 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " polyA_signal 4711..4832 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4748..4767) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4802..4821 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4881..4897) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4881..4897) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4894..4916) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4905..4921 /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(4929..4959) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4974..4995 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5112..5129) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5283..5871) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5363..5382) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6042..6902) /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 6665..6684 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6903..7007) /gene="bla" /label=AmpR promoter primer_bind complement(7082..7101) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata 601 gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg 661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc 721 tccggactct agcgtttaaa cttaagcttg gtaccgagct cggatccact agtccagtgt 781 ggtggaattc tgcagatatc cagcacagtg gcggccgctc gagaccatga aggacaacac 841 cgtgcccctg aagctgatcg ccctgctggc caacggcgag ttccactctg gcgagcagct 901 gggcgagaca ctgggcatga gcagagccgc catcaacaag cacatccaga ccctgcggga 961 ctggggcgtg gacgtgttca cagtgcccgg caagggctac agcctgcccg agcccatcca 1021 gctgctgaac gccaagcaga tcctgggcca gctggatggc ggatctgtgg ccgtgctgcc 1081 cgtgatcgac agcaccaacc agtatctgct cgaccggatc ggcgagctga agtccggcga 1141 cgcctgtatc gccgagtacc agcaggccgg cagaggcggc aggggcagaa agtggttcag 1201 ccccttcggc gccaacctgt acctgagcat gttctggaga ctggaacagg gcccagccgc 1261 cgctattggc ctgagcctgg tcatcggcat cgtgatggcc gaggtgctgc ggaagctggg 1321 cgccgacaaa gtgcgcgtga agtggcccaa cgacctgtat ctgcaggacc ggaagctggc 1381 cggcatcctg gtggaactga ccggcaaaac aggcgacgcc gcccagatcg tgatcggcgc 1441 tggcatcaac atggccatgc ggcgggtgga agagagcgtg gtcaaccagg gctggatcac 1501 cctgcaggaa gccggcatca acctggaccg gaacaccctg gccgccatgc tgatcagaga 1561 gctgagagcc gccctggaac tgttcgagca ggaaggcctg gccccctacc tgagcagatg 1621 ggagaagctg gacaacttca tcaaccggcc cgtgaaactg atcatcggcg acaaagagat 1681 cttcggcatc agccggggca tcgacaagca gggggccctg ctcctggaac aggacggcat 1741 catcaagccc tggatgggcg gcgagatcag cctgcggagc gccgagaagt acccatacga 1801 tgttcctgac tatgccggta ccgagctcgg atccaccatg gctagctgga gccacccgca 1861 gttcgagaaa ggtggaggtt ccggaggtgg atcgggaggt ggatcgtgga gccacccgca 1921 gttcgaaaaa gcggccgata tcacaagttt gtacaaaaaa gcaggctcca ccatgtacaa 1981 catgatggag acggagctga agccgccggg cccgcagcaa acttcggggg gcggcggcgg 2041 caactccacc gcggcggcgg ccggcggcaa ccagaaaaac agcccggacc gcgtcaagcg 2101 gcccatgaat gccttcatgg tgtggtcccg cgggcagcgg cgcaagatgg cccaggagaa 2161 ccccaagatg cacaactcgg agatcagcaa gcgcctgggc gccgagtgga aacttttgtc 2221 ggagacggag aagcggccgt tcatcgacga ggctaagcgg ctgcgagcgc tgcacatgaa 2281 ggagcacccg gattataaat accggccccg gcggaaaacc aagacgctca tgaagaagga 2341 taagtacacg ctgcccggcg ggctgctggc ccccggcggc aatagcatgg cgagcggggt 2401 cggggtgggc gccggcctgg gcgcgggcgt gaaccagcgc atggacagtt acgcgcacat 2461 gaacggctgg agcaacggca gctacagcat gatgcaggac cagctgggct acccgcagca 2521 cccgggcctc aatgcgcacg gcgcagcgca gatgcagccc atgcaccgct acgacgtgag 2581 cgccctgcag tacaactcca tgaccagctc gcagacctac atgaacggct cgcccaccta 2641 cagcatgtcc tactcgcagc agggcacccc tggcatggct cttggctcca tgggttcggt 2701 ggtcaagtcc gaggccagct ccagcccccc tgtggttacc tcttcctccc actccagggc 2761 gccctgccag gccggggacc tccgggacat gatcagcatg tatctccccg gcgccgaggt 2821 gccggaaccc gccgccccca gcagacttca catgtcccag cactaccaga gcggcccggt 2881 gcccggcacg gccattaacg gcacactgcc cctctcacac atgttggacc cagctttctt 2941 gtacaaagtg gtgacgtaag ctaggggccc gtttaaaccc gctgatcagc ctcgactgtg 3001 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa 3061 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt 3121 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa 3181 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc 3241 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt 3301 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 3361 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 3421 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 3481 tagggtgatg gttcacgtac ctagaagttc ctattccgaa gttcctattc tctagaaagt 3541 ataggaactt ccttggccaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg 3601 atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt 3661 gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat 3721 ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg 3781 gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca 3841 cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc 3901 gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca 3961 ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct 4021 gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg 4081 caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg 4141 cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt 4201 gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg 4261 aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag 4321 cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat 4381 cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca 4441 atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc 4501 gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact 4561 cgtccgaggg caaaggaata gcacgtacta cgagatttcg attccaccgc cgccttctat 4621 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg 4681 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac 4741 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 4801 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc 4861 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 4921 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 4981 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 5041 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 5101 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 5161 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 5221 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 5281 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 5341 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 5401 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 5461 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 5521 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 5581 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 5641 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 5701 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt 5761 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 5821 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 5881 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 5941 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 6001 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 6061 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 6121 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 6181 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 6241 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 6301 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 6361 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 6421 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 6481 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 6541 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 6601 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 6661 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 6721 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 6781 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 6841 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 6901 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 6961 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 7021 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca 7081 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg 7141 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca 7201 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca 7261 gatatacgcg tt //