LOCUS Exported 7305 bp DNA circular SYN 20-DEC-2021 DEFINITION . ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7305) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 18, 2022 from SnapGene Viewer 6.0.0 https://www.snapgene.com FEATURES Location/Qualifiers source 1..7305 /mol_type="other DNA" /organism="synthetic DNA construct" polyA_signal 576..710 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" /note="color: #a6acb3" enhancer 724..1103 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" /note="color: #ffffff" promoter 1105..1382 /label=chicken beta-actin promoter /note="color: #ffffff; direction: RIGHT" intron 1383..2399 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" /note="color: #------" regulatory 2454..2463 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /note="color: #a6acb3" /regulatory_class="other" CDS 2463..2486 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /note="color: #cc99b2" /translation="DYKDDDDK" CDS 2487..3446 /codon_start=1 /label=Sox2 WT /note="color: #993366" /translation="MYNMMETELKPPGPQQASGGGGGGGNATAAATGGNQKNSPDRVKR PMNAFMVWSRGQRRKMAQENPKMHNSEISKRLGAEWKLLSETEKRPFIDEAKRLRALHM KEHPDYKYRPRRKTKTLMKKDKYTLPGGLLAPGGNSMASGVGVGAGLGAGVNQRMDSYA HMNGWSNGSYSMMQEQLGYPQHPGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGS PTYSMSYSQQGTPGMALGSMGSVVKSEASSSPPVVTSSSHSRAPCQAGDLRDMISMYLP GAEVPEPAAPSRLHMAQHYQSGPVPGTAINGTLPLSHM" misc_feature 3462..4035 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /note="color: #a6acb3" misc_feature 3485..4037 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /note="color: #a6acb3" CDS 4037..4435 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /note="color: #ccffcc" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" protein_bind 4455..4488 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." /note="color: #31849b; direction: RIGHT" protein_bind 4761..4793 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." /note="color: #31849b; direction: RIGHT" primer_bind complement(4922..4938) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" /note="color: #a020f0; direction: LEFT" protein_bind 4946..4962 /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)." /note="color: #31849b" promoter complement(4970..5000) /label=lac promoter /note="promoter for the E. coli lac operon" /note="This reverse directional feature has 3 segments: 1: 4970 .. 4976 / #ffffff / -10 2: 4977 .. 4994 / #ffffff 3: 4995 .. 5000 / #ffffff / -35" protein_bind 5015..5036 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /note="color: #31849b" promoter 5095..5290 /label=SV40 promoter /note="SV40 early promoter" /note="color: #ffffff; direction: RIGHT" rep_origin 5141..5276 /label=SV40 ori /note="SV40 origin of replication" /note="color: #ffff00" polyA_signal 5296..5430 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" /note="color: #a6acb3" rep_origin complement(5669..6257) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" CDS complement(6428..7288) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /note="This feature has 2 segments: 1: 6428 .. 7219 / #ccffcc 2: 7220 .. 7288 / #ccffcc / signal sequence Cleavage site after base 7219" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" 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 attgtgctgt ctcatcattt tggcaaagaa ttagccacca 2461 tggattacaa ggatgacgac gataagatgt ataacatgat ggagacggag ctgaagccgc 2521 cgggcccgca gcaagcttcg gggggcggcg gcggaggagg caacgccacg gcggcggcga 2581 ccggcggcaa ccagaagaac agcccggacc gcgtcaagag gcccatgaac gccttcatgg 2641 tatggtcccg ggggcagcgg cgtaagatgg cccaggagaa ccccaagatg cacaactcgg 2701 agatcagcaa gcgcctgggc gcggagtgga aacttttgtc cgagaccgag aagcggccgt 2761 tcatcgacga ggccaagcgg ctgcgcgctc tgcacatgaa ggagcacccg gattataaat 2821 accggccgcg gcggaaaacc aagacgctca tgaagaagga taagtacacg cttcccggag 2881 gcttgctggc ccccggcggg aacagcatgg cgagcggggt tggggtgggc gccggcctgg 2941 gtgcgggcgt gaaccagcgc atggacagct acgcgcacat gaacggctgg agcaacggca 3001 gctacagcat gatgcaggag cagctgggct acccgcagca cccgggcctc aacgctcacg 3061 gcgcggcaca gatgcaaccg atgcaccgct acgacgtcag cgccctgcag tacaactcca 3121 tgaccagctc gcagacctac atgaacggct cgcccaccta cagcatgtcc tactcgcagc 3181 agggcacccc cggtatggcg ctgggctcca tgggctctgt ggtcaagtcc gaggccagct 3241 ccagcccccc cgtggttacc tcttcctccc actccagggc gccctgccag gccggggacc 3301 tccgggacat gatcagcatg tacctccccg gcgccgaggt gccggagccc gctgcgccca 3361 gtagactgca catggcccag cactaccaga gcggcccggt gcccggcacg gccattaacg 3421 gcacactgcc cctgtcgcac atgtgagaat tccgcccccc ccccctctcc ctcccccccc 3481 cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta 3541 ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc 3601 ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat 3661 gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc 3721 ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt 3781 gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt 3841 gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag 3901 aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt 3961 tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga 4021 aaaacacgat gataatatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag 4081 agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc 4141 tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc 4201 ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg 4261 tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca 4321 ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc 4381 gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctaagagct 4441 cgtcgaggga cctaataact tcgtatagca tacattatac gaagttatac atgtttaagg 4501 gttccggttc cactaagtac aattcgatat caagcttatc gatgatctag aactcgctga 4561 tcagcctcga ctgggccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct 4621 tccttgaccc tggaaggggc catcccactg tcctttccaa aaaaatggga aattgcatcc 4681 attgtcgaga aggttcatct aatctggggg gtgggggtgg gaagacccaa ggggagaaat 4741 gggaaacaat aacggccttt taacttcgta tagcatacat tatacgaagt tatacatgtt 4801 taagggttcc ggttccacta agtacaattc gatatcaagc ttgcaggcat gctggggatg 4861 cggtgggctc tatggcttct gaggcggaaa gaacctgcag cccaagcttg gcgtaatcat 4921 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 4981 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 5041 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct 5101 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc 5161 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga 5221 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg 5281 cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc 5341 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 5401 ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg 5461 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct 5521 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca 5581 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga 5641 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 5701 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 5761 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 5821 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt 5881 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 5941 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 6001 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 6061 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg 6121 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 6181 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 6241 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 6301 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 6361 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 6421 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 6481 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 6541 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 6601 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 6661 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 6721 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 6781 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 6841 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 6901 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 6961 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 7021 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 7081 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 7141 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 7201 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 7261 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaat //