LOCUS Exported 5400 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Homology arms and mEGFP-linker sequence for N-terminus tagging of human SON. ACCESSION . VERSION . KEYWORDS AICSDP-82:SON-mEGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5400) TITLE Allen Institute for Cell Sciences Plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 5400) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5400 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(61..649) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(141..160) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(820..1680) /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 1443..1462 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1681..1785) /gene="bla" /label=AmpR promoter primer_bind 1853..1871 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(1909..1931) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 2031..2050 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2244..2266 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2258..2275 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2259..2275 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS 3293..4009 /codon_start=1 /product="enhanced GFP with monomerizing A206K mutation (Zacharias et al., 2002)" /label=mEGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3338..3359) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3599..3618) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3946..3967 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(5059..5075) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5059..5075) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5072..5094) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5083..5099 /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(5107..5137) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5152..5173 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5290..5307) /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 61 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 121 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 181 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 241 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 301 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 361 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 421 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 481 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac 541 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 601 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 661 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 721 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 781 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 841 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 901 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 961 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 1021 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 1081 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 1141 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 1201 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 1261 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 1321 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 1381 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 1441 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 1501 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 1561 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 1621 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 1681 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 1741 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 1801 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 1861 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 1921 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 1981 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 2041 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 2101 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 2161 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 2221 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt 2281 gacgcgtatt gggatcccgc tcgccggaag tcggcccacc gggaccggcg cgggaccagc 2341 gcggcaccgc ccctcactgc cccgccctac gacggcttcc ctgccccatt ggtccgcgcg 2401 gccctgggtc tccgcggcga gaatctagtc tgattccctc ccgccctggg cgctgcctga 2461 ctgctcccca gccgagacac ccaagcctta gcacgtttat aatgtgacag tggtacgggg 2521 tgccacgcag tgtttccagg agcgccgggg gggagcgcca aaatgctaaa cccattctta 2581 gccaattgtg cgccgctcac gagtcagagc tgagcagcta ccgcggaaaa atctcgtgac 2641 acgcctgact cctgatgaga ctaaaatgca ccgggctttg aatgccggtc gcttaggaca 2701 tccggcctat tgctaaaaaa aaaaatgagg ctggttatag aacgcactgg ggaagacaca 2761 gggcaacagc gagttcctcc tctcacatgc cagtgggggc gggggctgaa aaattctctc 2821 ccggatgcat atccctgtcg aaactgaact tggcgccacc tgagagcgcc tgaaagccgt 2881 gtgttaaaca ccgggttgcc cttcttgctc ttcttcctct tcgcctttaa tctcaggcac 2941 ttagcaagcc tgttaaggct cccaaacact gctggggact gacaacttat gcggacacgg 3001 tcgcctcaag agagacggct cctgtaatgg cgcctgcgtc agacactaac atggcggaca 3061 tagtcgtgcg cggcgcaaac gtcagcacct ctaccccagc cgctcagaaa cagcagagga 3121 gagcagggcc gtgctcactg attggtccct cggtggagcg ggccggaccg ggcattccct 3181 ttggagccaa tcaggaaggc gagagaccga attggccgct gcgcctcctc ccgaggcatg 3241 ctgggagcct ggaggactag cgaggaggag ttgagagaac ggagcggacg ccatggtgag 3301 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt 3361 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct 3421 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac 3481 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga 3541 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga 3601 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg 3661 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga 3721 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa 3781 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta 3841 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag 3901 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga 3961 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaaga gcggcctgag 4021 aagcgcgaca aacatcgagc agatctttag gtctttcgtc gtcagtaaat tccgggaaat 4081 tcaacaggag ctttccaggt aaacgcctcc cagattcttc tgctcccaac ggaccgttag 4141 gccctcacct agccttcttt ggcacctttc ttcggagacg cagtcgttcc ccggctcagg 4201 ccccgctagg gccccgcctg ggcctgggat ccattttccg ggcccccccc aaccgccccc 4261 ccagccgctc ccttcctttt ccctcgagga acttcgcctt ttccgaggta acccaagact 4321 ccctgtttca gaaccgagtt aagatgttca cccttctccc ctgtttgggt cccccctgcc 4381 gtttagctac tcgtaggccc ggttgtccgc cagggacggg cctagttcct tcctcagctc 4441 ctcctccgcc gccgcgtatc ccctccccct ctccgcctcc gcggattttg ccggccggtg 4501 cctatgaccg gaaagggtct ctaacggccg tttcttggtt aatcttaaat cctctttatt 4561 attgcgagct tcagaccgct tgggggtctg gatctattaa taagattatt cggggcctag 4621 tgtcctgtag tatctgtcag ccgttctgat agttaaatgt ttcaattaaa acgaactcaa 4681 actgatgcac ttgctagttt gttaaaatac tcgagataaa gatggaaggg gactttgaaa 4741 tggttgcgat taattacagc aagtgaaagt ggactttgtg tgtaggtcta ttattttctt 4801 gctacccctg ccccagttga cacgctattt ttgtcattgc tggtagactt tacataattt 4861 gcgcttggct tcacaaaatt tgaaggcgaa gagattgtta aagaaatggt tgtttactta 4921 ggaaaatagt gtgtcatgtt cagcggttgt ctctcgatat ttgcttttgt ataagatact 4981 catttcttta aagtacagaa gtaggaaaga gatcttgaaa cgtaatccca atggcgcgcc 5041 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 5101 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 5161 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 5221 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 5281 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 5341 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa //