LOCUS Exported 7242 bp ds-DNA circular SYN 29-MAR-2022 DEFINITION expression of Sox2. ACCESSION . VERSION . KEYWORDS pCAG-FLAG-IRES-blasticidin-Sox2 WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7242) TITLE Jang Lab Plasmids 2022 REFERENCE 2 (bases 1 to 7242) AUTHORS . TITLE Direct Submission JOURNAL Exported Mar 29, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7242 /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" regulatory 2454..2463 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2463..2486 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" misc_feature 3479..4052 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" misc_feature 3502..4054 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(3646..3663) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 3873..3892 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 4054..4452 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" protein_bind 4472..4505 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind complement(4582..4599) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4588..4812 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(4859..4875) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4859..4875) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4872..4894) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4883..4899 /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(4907..4937) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4952..4973 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5032..5227 /label=SV40 promoter /note="SV40 early promoter" rep_origin 5078..5213 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5140..5159 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 5233..5367 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5435..5452) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5606..6194) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5686..5705) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6365..7225) /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 6988..7007 /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 cgccrggsgg ggsggggsgg ggsgrggggs 1261 ggggsggggs grggcggaga 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 grgggctcgg gggrggggcg 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 ttcctacagc 2401 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttagccacca 2461 tggattacaa ggatgacgac gataagctcg agaccatgta taacatgatg gagacggagc 2521 tgaagccgcc gggcccgcag caagcttcgg ggggcggcgg cggaggaggc aacgccacgg 2581 cggcggcgac cggcggcaac cagaagaaca gcccggaccg cgtcaagagg cccatgaacg 2641 ccttcatggt atggtcccgg gggcagcggc gtaagatggc ccaggagaac cccaagatgc 2701 acaactcgga gatcagcaag cgcctgggcg cggagtggaa acttttgtcc gagaccgaga 2761 agcggccgtt catcgacgag gccaagcggc tgcgcgctct gcacatgaag gagcacccgg 2821 attataaata ccggccgcgg cggaaaacca agacgctcat gaagaaggat aagtacacgc 2881 ttcccggagg cttgctggcc cccggcggga acagcatggc gagcggggtt ggggtgggcg 2941 ccggcctggg tgcgggcgtg aaccagcgca tggacagcta cgcgcacatg aacggctgga 3001 gcaacggcag ctacagcatg atgcaggagc agctgggcta cccgcagcac ccgggcctca 3061 acgctcacgg cgcggcacag atgcaaccga tgcaccgcta cgacgtcagc gccctgcagt 3121 acaactccat gaccagctcg cagacctaca tgaacggctc gcccacctac agcatgtcct 3181 actcgcagca gggcaccccc ggtatggcgc tgggctccat gggctctgtg gtcaagtccg 3241 aggccagctc cagccccccc gtggttacct cttcctccca ctccagggcg ccctgccagg 3301 ccggggacct ccgggacatg atcagcatgt acctccccgg cgccgaggtg ccggagcccg 3361 ctgcgcccag tagactgcac atggcccagc actaccagag cggcccggtg cccggcacgg 3421 ccattaacgg cacactgccc ctgtcgcaca tgtgagcggc cgcgaattcc gccccccccc 3481 cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg 3541 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga 3601 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa 3661 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa 3721 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct 3781 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg 3841 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg 3901 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca 3961 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg 4021 acgtggtttt cctttgaaaa acacgatgat aatatggcca agcctttgtc tcaagaagaa 4081 tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac 4141 agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat 4201 cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca 4261 gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc 4321 tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag 4381 gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg 4441 tgggagggct aagaattcgt cgagggacct aataacttcg tatagcatac attatacgaa 4501 gttatacatg tttaagggtt ccggttccac taggtacaat tcgatatcaa gcttatcgat 4561 gatctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt 4621 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat 4681 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg 4741 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg 4801 tgggctctat ggcttctgag gcggaaagaa cctgcagccc aagcttggcg taatcatggt 4861 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 4921 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 4981 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa 5041 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag 5101 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc 5161 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt 5221 ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 5281 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 5341 atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa cgcgcgggga 5401 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 5461 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 5521 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 5581 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 5641 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 5701 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 5761 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 5821 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 5881 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 5941 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 6001 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 6061 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 6121 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 6181 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 6241 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 6301 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 6361 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 6421 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 6481 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 6541 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 6601 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 6661 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 6721 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 6781 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 6841 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 6901 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 6961 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 7021 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 7081 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 7141 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 7201 cgacacggaa atgttgaata ctcatactct tcctttttca at //