LOCUS Exported 7441 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor ZNF211 with GFP. ACCESSION . VERSION . KEYWORDS pZNF211-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7441) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 7441) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7441 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(34..50) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(34..50) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(47..69) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 58..74 /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(82..112) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 127..148 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(265..282) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(436..1024) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(516..535) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1195..2055) /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 1818..1837 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2056..2160) /gene="bla" /label=AmpR promoter primer_bind 2228..2246 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(2284..2306) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 2406..2425 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2619..2641 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2633..2650 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2634..2650 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS 3887..3907 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3920..3964 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 4004..4027 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 4055..4774 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK EFVTAAGITLGMDELYK" primer_bind complement(4100..4121) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4361..4380) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4812..5397 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4978..4995) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5205..5224 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5479..6282 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5542..5561) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6152..6171 /label=Neo-F /note="Neomycin resistance gene, forward primer" ORIGIN 1 gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg 61 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg 121 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc 181 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt 241 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct 301 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga 361 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc 421 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg 481 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg 541 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt 601 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt 661 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg 721 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact 781 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt 841 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct 901 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac 961 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc 1021 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg 1081 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta 1141 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca 1201 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc 1261 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc 1321 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc 1381 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat 1441 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt 1501 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc 1561 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag 1621 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt 1681 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac 1741 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg 1801 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat 1861 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc 1921 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc 1981 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa 2041 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg 2101 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg 2161 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac 2221 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga 2281 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg 2341 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa 2401 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca 2461 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg 2521 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg 2581 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac 2641 gacggccagt gaattcgagc tcggtaccca tgtggacact gcctcgttta cacagagttg 2701 catagtccat gtgtcggaga aaccctttac ctgcagggag atcaggaaag acttcctggc 2761 caacatgagg tttctccatc aagacgccac tcaaacaggg gagaagccaa ataacagtaa 2821 caagtgtgcg gtggcctttt acagtggaaa aagtcatcac aactggggaa aatgcagtaa 2881 agcctttagc cacatagaca cacttgttca ggaccagaga atcctcacta gagaaggact 2941 ttttgagtgc agtaaatgtg ggaaagcatg tacgcgaaga tgtaacctca ttcagcacca 3001 gaaagtccac agtgaagaaa ggccttatga atgcaatgaa tgtggaaaat tctttaccta 3061 ctactccagt ttcattatac atcagagagt tcatactgga gaaaggcctt atgcgtgccc 3121 tgaatgtggg aaatcgttta gtcagatata cagcctcaat agccatagga aagttcacac 3181 tggagaaagg ccttatgaat gtggggaatg tgggaaatct tttagccaaa ggtccaacct 3241 catgcagcat cgcagagttc acactggaga aaggccttat gaatgcagcg aatgtgggaa 3301 atcttttagc caaaacttta gcctgatcta ccaccagaga gttcacactg gagaaagacc 3361 tcatgagtgc aatgaatgtg gaaaatcctt tagccgaagc tccagcctca ttcaccaccg 3421 gagacttcac actggagaaa gaccctatga gtgcagtaaa tgtgggaagt catttaagca 3481 aagctccagc ttcagttcac atcggaaagt ccacacaggg gaaaggcctt atgtgtgtgg 3541 ggaatgtggg aaatccttta gccatagctc caaccttaag aaccaccaga gagttcacac 3601 tggagaaaga cctgttgagt gcagtgaatg tagcaaatcc tttagctgta aatctaacct 3661 cattaaacac ctgagagttc acactggaga aaggccttat gagtgcagtg aatgtgggaa 3721 atcctttagc caaagttcta gcctcattca acaccgcaga gttcacacgg gaaaaaggcc 3781 ttatcagtgc agtcaatgtg ggaaatcctt tggctgcaaa tctgtcctca ttcaacacca 3841 gagagttcac attggagaaa agcccgggat tccaactact gcaagcgaga atctttattt 3901 tcagggcgcc gccaaattca aagaaaccgc tgctgctaaa ttcgaacgcc agcacatgga 3961 cagcggaggt ggaggttcga gcggtccctc gggttcgtcg agcctggaag ttctgttcca 4021 ggggcccctg tcgtcgagcg gtccctcggg ttcgatggtg agcaaaggcg aggagctgtt 4081 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag 4141 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg 4201 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt 4261 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat 4321 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac 4381 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat 4441 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca 4501 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg 4561 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat 4621 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag 4681 caaagacccc aacgagaagc gcgatcacat ggtccttaag gagttcgtga ccgccgccgg 4741 gatcactctc ggcatggacg agctgtacaa gtaatgatct agagtcgagt taattaagaa 4801 ttccgccccc ccccctctcc ctcccccccc ctaacgttac tggccgaagc cgcttggaat 4861 aaggccggtg cgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg 4921 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc 4981 tcgccaaagg aatgcagggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt 5041 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg 5101 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac 5161 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg 5221 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg 5281 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc 5341 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaacctgt 5401 tacattgcac aagataaaaa tatatcatca cgaacagtaa aactgtctgc ttacataaac 5461 agtaatacaa ggggtgttat gggatcggcc attgaacaag atggattgca cgcaggttct 5521 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc 5581 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc 5641 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc 5701 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg 5761 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag 5821 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc 5881 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt 5941 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc 6001 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc 6061 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg 6121 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag 6181 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg 6241 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagtcctgta ctgagaattg 6301 caatttcctt ttagtgtaat tatactgaag gagtacacct gtgagagaga caagtacctg 6361 atttggaagc cccaacatct aaggatatac agtgggcgga ttccccttaa gttccaggta 6421 tgtgttacac tttctaacat gccatttaga aagtgttaga ctttctcacc tgccatttat 6481 ggctcttgcc gtttatgtca ctgacagttt ctgaggcaga agccgtatca tgtctaccac 6541 ctgtgaggtc cacacagtgt gtatcattta cctcctgaac ctgctcaagg aagcagacct 6601 ctgcttctcc ccatttgcta gaagaaatca tgaatagtct gagtcttcct ctctgacaag 6661 ttagggcatg gacttgaccc agctttgtgc cagagaaccc aatatgagtg ttgttggcag 6721 cttgccaaga aggactgtct ttttcaagac atactggttt catgtgacac ctccatggat 6781 ttttttccag cctctaagtc accaacttgg gaactgcttg tctcacgttg ctttgttttt 6841 tacaataata aaagcattat tatttaagct acctttatct tggcatttga ttcactcttt 6901 gaggtggttc aaaaacagaa tcgggccctg ccagctttac agagtaaaca cctttggagg 6961 gagagggagg tgtcatatgc ctcagtgaca acagtataat ggaaaaatac aactgtcttt 7021 ccagctttgg cctaatctga atttgaatta ttgcccaagg ccttctagga aatactgcag 7081 gatattatgg tcttaattcg tggactggat gcaaggattt tttgtgggct caaaggacac 7141 cctgaggaag tgggaaattg tgacaaattc cattgcttct gggaacaaga tgaattttgt 7201 tgttattcac aagggttatt acatactgcc cagcactgtt gaggcagtct acccccagtg 7261 ctgccaagga gccaagtgcc ccaatacaat ttagtaggct actgtcactg taaaatgatg 7321 ggggaaccat aattggtgtg gaaacactgg gttactaggg agtgaaggag actgcagcaa 7381 actagatgga atgtgctcct tgtaatatgt tctagtaatt gtgcaggatc agtgtgtaca 7441 g //