LOCUS Exported 6743 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor ZNF584 with GFP. ACCESSION . VERSION . KEYWORDS pZNF584-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6743) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 6743) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6743 /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 3567..3587 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3600..3644 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3684..3707 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 3735..4454 /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(3780..3801) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4041..4060) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4492..5077 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4658..4675) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4885..4904 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5159..5962 /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(5222..5241) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5832..5851 /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 tcggtacccc caagaaggca cactgagcat ggggcagcct 2701 tcccacctgg ttccagttgt gggcaacagc aagaagtcca tgtggcagag aagctgttca 2761 aatgcagtga ctgtgggaag gtgttcttaa aggcctttgc cctccttgac catctgataa 2821 cgcattctga agagagaccc ttcagatgcc caacaggcag aagtgctttc aagaagtcag 2881 ctcatattaa cccccgaaaa attcacactg gagaaacagc ccatgtgtgt aatgaatgtg 2941 ggaaggcctt cagttacccg tctaagctga ggaaacacca gaaggttcac acaggcataa 3001 aaccttttaa gtgtagtgac tgtggtaaaa ccttcaaccg caaagacgca cttgttctac 3061 accagaggat tcacactgga gaaaggcctt acgagtgcag caaatgtggt aaaaccttca 3121 gtgttctgtc taccctcatt cggcaccgga aagtgcacat tggagaaagg ccctatgagt 3181 gtgcagaatg tgggaagttc tttaaataca ataatagctt cattcttcac cagagagttc 3241 acactggaga aaggcctttt gaatgcaagc aatgtgggaa aggctacgtg acccgttcag 3301 gcctctatca gcactggaaa gtccacactg gggaacggcc ctatgaatgt agcctgtgtg 3361 ggaaaacctt cactaccaga tcctaccgca atcggcacca gcagttccac actgaagaga 3421 ggtcttatga atgtacagag tgtgggaaag ccttcaaaca tagttccacc ctccttcagc 3481 acaagaaagt ccatactcca gaaaggcgtc aggaggacag ggcacatggg aaggtcgtta 3541 gctgcgggat tccaactact gcaagcgaga atctttattt tcagggcgcc gccaaattca 3601 aagaaaccgc tgctgctaaa ttcgaacgcc agcacatgga cagcggaggt ggaggttcga 3661 gcggtccctc gggttcgtcg agcctggaag ttctgttcca ggggcccctg tcgtcgagcg 3721 gtccctcggg ttcgatggtg agcaaaggcg aggagctgtt caccggggtg gtgcccatcc 3781 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg 3841 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg 3901 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc 3961 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg 4021 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg 4081 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca 4141 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg 4201 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca 4261 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc 4321 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc 4381 gcgatcacat ggtccttaag gagttcgtga ccgccgccgg gatcactctc ggcatggacg 4441 agctgtacaa gtaatgatct agagtcgagt taattaagaa ttccgccccc ccccctctcc 4501 ctcccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc 4561 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc 4621 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcagggt 4681 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct 4741 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa 4801 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt 4861 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag 4921 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt 4981 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt 5041 ttcctttgaa aaacacgatg ataatatggc cacaacctgt tacattgcac aagataaaaa 5101 tatatcatca cgaacagtaa aactgtctgc ttacataaac agtaatacaa ggggtgttat 5161 gggatcggcc attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag 5221 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg 5281 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa 5341 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc 5401 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc 5461 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga 5521 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa 5581 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct 5641 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat 5701 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt 5761 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta 5821 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga 5881 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg 5941 ccttcttgac gagttcttct gagtcgagca ccgtgttcat caggaaaggt cttattccag 6001 aaaggaggtt gaggagagtg gccgtgagag tgccatccga aagaagctaa accttgcaca 6061 tcccaacacc caccccaggg agagttcccg tgtgtgcctg gtgtgtggga cgctttcggg 6121 agccacattg cactctgact tgcctggggc tgttggcagt gtcacatcac tgagtttatc 6181 caccgccatc cacctctatc caccccataa ggtccctaca gcaagtggga cagcagacct 6241 tgtgctcctt gctctaaaca gtagagaatc gttaatagtg gagcccaaaa caggcctcat 6301 tccctaccct tgactggttt agctgtggac atgacccacc tctggccgga ggagctgagc 6361 tggtatctgc tgggggcttc ctgggaaggt ttaccatttt catggaaatg gttgacatta 6421 cacatggtat ttgtcatgcc ctagcatgga ctgtctcata ctgctctgga ctatgtggta 6481 ataaaggctt tattgtttaa gccgcttaaa attttttttt tatttttttt ttgagatggt 6541 cttattctgt tgcccaggct ggaatgcagt ggtgccatca tagctcacca cagccttgaa 6601 ctgctggtct caagggatct tcctgcttca gcctcctgag tagctggcac tacaggtgtg 6661 cgccatcaca cctggctaat tttttttttt catttgtgga cagggtcttg ctatattgcc 6721 caagctggtc tcgaactctg agc //