LOCUS Exported 7267 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor ZNF347 with GFP. ACCESSION . VERSION . KEYWORDS pZNF347-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7267) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 7267) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7267 /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 3881..3901 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3914..3958 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3998..4021 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 4049..4768 /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(4094..4115) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4355..4374) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4806..5391 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4972..4989) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5199..5218 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5473..6276 /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(5536..5555) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6146..6165 /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 tcggtacccg cctagctatt catctggtaa ttcacaccgg 2701 agaaaagcct tacaaatgtc atgaatgcgg caaggtcttt aggcgtaatt cacaccttgc 2761 aaggcatcag ctaattcata ctggagagaa accttataag tgtaatgagt gtggcaaagc 2821 ctttagagca cattcaaacc taactaccca tcaggtcatc catactggag aaaagcctta 2881 caaatgtaat gaatgtggca aggtcttcac tcaaaattca caccttgcaa atcatcaaag 2941 aattcatact ggagtgaagc cttatatgtg taatgagtgc ggcaaagcct tcagtgtgta 3001 ttcaagccta actacccatc aggtcatcca tactggagaa aaaccttaca aatgtaatga 3061 gtgtggcaag gtcttcactc agaattcaca ccttgcaaga catcggggaa ttcatactgg 3121 agagaaacct tataaatgta atgaatgtgg caaggtcttt aggcataatt catacctttc 3181 aaggcatcag cgaattcata ctggagagaa accttacaag tataatgagt atggcaaagc 3241 ctttagtgaa cattcaaacc taactaccca tcaggtcatc catactggtg aaaaacctta 3301 caaatgtaat gagtgtggca aggtcttcac tcagaattca caccttgcaa gacatcggag 3361 agttcatact ggaggtaaac cttaccagtg taatgaatgt ggcaaagcct ttagtcaaac 3421 atcaaagctt gcaaggcatc agagagttca tactggagag aaaccatatg agtgtaatca 3481 gtgtgggaaa gcctttagtg tccgttcaag cctaactacc catcaggcaa tccatactgg 3541 aaaaaaacct tacaaatgca atgagtgtgg gaaggtcttc actcagaatt cacaccttgc 3601 aagacatcgg ggaattcata ctggagagaa accttacaag tgtaatgagt gtggcaaagc 3661 ctttagtcaa acttcaaaac ttgcaaggca tcagagaatt cataccggag agaaaccata 3721 tgagtgtggg aaacccttta gtatctgttc aagcctaact acccatcaga caatccatac 3781 tggtgggaaa ccttacaaat gtaacgtgtg gaaagttcta aagtcagagt tcaaaccttg 3841 caagccatca cagaattcag ggattccaac tactgcaagc gagaatcttt attttcaggg 3901 cgccgccaaa ttcaaagaaa ccgctgctgc taaattcgaa cgccagcaca tggacagcgg 3961 aggtggaggt tcgagcggtc cctcgggttc gtcgagcctg gaagttctgt tccaggggcc 4021 cctgtcgtcg agcggtccct cgggttcgat ggtgagcaaa ggcgaggagc tgttcaccgg 4081 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc 4141 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac 4201 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg 4261 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga 4321 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc 4381 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt 4441 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt 4501 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa 4561 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga 4621 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga 4681 ccccaacgag aagcgcgatc acatggtcct taaggagttc gtgaccgccg ccgggatcac 4741 tctcggcatg gacgagctgt acaagtaatg atctagagtc gagttaatta agaattccgc 4801 ccccccccct ctccctcccc ccccctaacg ttactggccg aagccgcttg gaataaggcc 4861 ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg 4921 cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca 4981 aaggaatgca gggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa 5041 gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt 5101 gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt 5161 gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca 5221 acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc 5281 ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc 5341 acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaac ctgttacatt 5401 gcacaagata aaaatatatc atcacgaaca gtaaaactgt ctgcttacat aaacagtaat 5461 acaaggggtg ttatgggatc ggccattgaa caagatggat tgcacgcagg ttctccggcc 5521 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat 5581 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg 5641 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg 5701 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta 5761 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta 5821 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc 5881 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc 5941 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg 6001 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg 6061 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt 6121 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc 6181 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc 6241 atcgccttct atcgccttct tgacgagttc ttctgagtct ggagagaaac cttacaaatg 6301 aaggtggtaa cttttgcagt tagaattcat tcctttgaca acgtcagaga atttatgcct 6361 gagagattcc ttgaaaatgc aatgactggc tgggtgcagt ggctcatgcc tgtaatccca 6421 acactttggg agtccgaggc aggtggatca cctgaggtca ggagttcaag accagcctga 6481 ccaacatggc gaaaccccat ttctactaaa agtacacaat tagctaggcc tggtggcgtg 6541 cgcctgtaat cccagctata taggaggctg aggcaggaga atcacttgaa cccaggaggc 6601 ggaggttgca gtgagccaag atcgtgccaa tgcactccat cctgggcaaa aagagcaaaa 6661 ctccatctta aaggaaaaaa aaaaaatgca atgactgtgg caaactcctc atgagttcaa 6721 gtattaatag acatccaaga gtccataata atgagaaatc acataaatgt actgtgtgtg 6781 acagaggctt tattcaggcc tcacagcttt gcaggcatta aaatctacgt atttgatgga 6841 accacacaaa tgtaatgtgt aagagcatcc tgaagctctt actctaagac tgtaactgta 6901 ggtgaggatt catacaaaga gtgactcagt ctctagttct ttgattcact tttaaaatga 6961 taaactgcac aataattaca cgttccaata tggtaaaact taagacatta tatatttcaa 7021 aggtcaaagg acatttggaa atggctactt accttcaggt tataaaatct tttattccat 7081 tgtttgaggt ttctcaaaaa ggctactgtt tgtgactttc aacctgaact atggatctgt 7141 ggctattttg catccttaag cacctttcgt ggtgcctatc tgggaaagaa ccacagtatg 7201 gaaaggctta cttcattaca tgaggatcat ttttatccat gtttattgaa gaaggaggac 7261 tctgcct //