LOCUS Exported 5418 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Homology arms and linker-mEGFP sequence for C-terminus tagging of human SOX2. ACCESSION . VERSION . KEYWORDS AICSDP-60: SOX2-mEGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5418) AUTHORS Allen Institute for Cell Science TITLE Allen Institute for Cell Science Plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 5418) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5418 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 40..58 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(96..118) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 218..237 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 431..453 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 445..462 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 446..462 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS 1516..2232 /codon_start=1 /product="enhanced GFP with monomerizing A206K mutation (Zacharias et al., 2002)" /label=mEGFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(1558..1579) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1819..1838) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2166..2187 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(3264..3280) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3264..3280) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3277..3299) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3288..3304 /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(3312..3342) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3357..3378 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3495..3512) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3666..4254) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3746..3765) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4425..5285) /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 5048..5067 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5286..5390) /gene="bla" /label=AmpR promoter ORIGIN 1 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc 61 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag 121 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca 181 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg 241 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc 301 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct 361 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg 421 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgac gcgtattggg 481 attgaattga cgcgtattgg gatgccgagg gtcggcggcc gccggcgggc cgggcccgcg 541 cacagcgccc gcatgtacaa catgatggag acggagctga agccgccggg cccgcagcaa 601 acttcggggg gcggcggcgg caactccacc gcggcggcgg ccggcggcaa ccagaaaaac 661 agcccggacc gcgtcaagcg gcccatgaat gccttcatgg tgtggtcccg cgggcagcgg 721 cgcaagatgg cccaggagaa ccccaagatg cacaactcgg agatcagcaa gcgcctgggc 781 gccgagtgga aacttttgtc ggagacggag aagcggccgt tcatcgacga ggctaagcgg 841 ctgcgagcgc tgcacatgaa ggagcacccg gattataaat accggccccg gcggaaaacc 901 aagacgctca tgaagaagga taagtacacg ctgcccggcg ggctgctggc ccccggcggc 961 aatagcatgg cgagcggggt cggggtgggc gccggcctgg gcgcgggcgt gaaccagcgc 1021 atggacagtt acgcgcacat gaacggctgg agcaacggca gctacagcat gatgcaggac 1081 cagctgggct acccgcagca cccgggcctc aatgcgcacg gcgcagcgca gatgcagccc 1141 atgcaccgct acgacgtgag cgccctgcag tacaactcca tgaccagctc gcagacctac 1201 atgaacggct cgcccaccta cagcatgtcc tactcgcagc agggcacccc tggcatggct 1261 cttggctcca tgggttcggt ggtcaagtcc gaggccagct ccagcccccc tgtggttacc 1321 tcttcctccc actccagggc gccctgccag gccggggacc tccgggacat gatcagcatg 1381 tatctccccg gcgccgaggt gccggaaccc gccgccccca gcagacttca catgtcccag 1441 cactaccaga gcggcccggt gcccggcacg gccattaacg gcacactgcc cctctcacac 1501 atgggcggaa gcggagtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 1561 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 1621 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 1681 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc 1741 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 1801 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 1861 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 1921 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac 1981 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc 2041 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 2101 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc 2161 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 2221 ctgtacaagt gagggccgga cagcgaactg gaggggggag aaattttcaa agaaaaacga 2281 gggaaatggg aggggtgcaa aagaggagag taagaaacag catggagaaa acccggtacg 2341 ctcaaaaaga aaaaggaaaa aaaaaaatcc catcacccac agcaaatgac agctgcaaaa 2401 gagaacacca atcccatcca cactcacgca aaaaccgcga tgccgacaag aaaactttta 2461 tgagagagat cctggacttc tttttggggg actatttttg tacagagaaa acctggggag 2521 ggtggggagg gcgggggaat ggaccttgta tagatctgga ggaaagaaag ctacgaaaaa 2581 ctttttaaaa gttctagtgg tacggtagga gctttgcagg aagtttgcaa aagtctttac 2641 caataatatt tagagctagt ctccaagcga cgaaaaaaat gttttaatat ttgcaagcaa 2701 cttttgtaca gtatttatcg agataaacat ggcaatcaaa atgtccattg tttataagct 2761 gagaatttgc caatattttt caaggagagg cttcttgctg aattttgatt ctgcagctga 2821 aatttaggac agttgcaaac gtgaaaagaa gaaaattatt caaatttgga cattttaatt 2881 gtttaaaaat tgtacaaaag gaaaaaatta gaataagtac tggcgaacca tctctgtggt 2941 cttgtttaaa aagggcaaaa gttttagact gtactaaatt ttataactta ctgttaaaag 3001 caaaaatggc catgcaggtt gacaccgttg gtaatttata atagcttttg ttcgatccca 3061 actttccatt ttgttcagat aaaaaaaacc atgaaattac tgtgtttgaa atattttctt 3121 atggtttgta atatttctgt aaatttattg tgatatttta aggttttccc ccctttattt 3181 tccgtagttg tattttaaaa gattcggctc tgtattattt gaatcagtca tcccaatggc 3241 gcgccgagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 3301 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 3361 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 3421 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 3481 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 3541 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 3601 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 3661 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 3721 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 3781 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 3841 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 3901 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 3961 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 4021 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 4081 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca 4141 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 4201 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 4261 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 4321 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 4381 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 4441 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 4501 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 4561 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 4621 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 4681 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 4741 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 4801 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 4861 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 4921 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 4981 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 5041 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 5101 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 5161 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 5221 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 5281 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 5341 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 5401 cgaaaagtgc cacctgac //