LOCUS Exported 6769 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor ZNF696 with GFP. ACCESSION . VERSION . KEYWORDS pZNF696-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6769) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 6769) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6769 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(19..36) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(19..35) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(28..50) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(244..263) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 363..385 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(423..441) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 509..613 /gene="bla" /label=AmpR promoter CDS 614..1474 /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 complement(832..851) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1645..2233 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2134..2153 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2387..2404 /label=L4440 /note="L4440 vector, forward primer" protein_bind 2521..2542 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2557..2587 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2595..2611 /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)." primer_bind 2600..2622 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2619..2635 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2619..2635 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 3528..3548 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3561..3605 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3645..3668 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 3696..4415 /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(3741..3762) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4002..4021) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4453..5038 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4619..4636) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4846..4865 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5120..5923 /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(5183..5202) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5793..5812 /label=Neo-F /note="Neomycin resistance gene, forward primer" ORIGIN 1 ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg 61 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga 121 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct 181 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct 241 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc 301 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt 361 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa 421 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac 481 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 541 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 601 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc 661 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga 721 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga 781 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg 841 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc 901 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac 961 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact 1021 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca 1081 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg 1141 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact 1201 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg 1261 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg 1321 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat 1381 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc 1441 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat 1501 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt 1561 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc 1621 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt 1681 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac 1741 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt 1801 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 1861 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 1921 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 1981 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 2041 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt 2101 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc 2161 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg 2221 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc 2281 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc 2341 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag 2401 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca 2461 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat 2521 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg 2581 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga 2641 ttacgccaag cttggtcact tctgagaaaa ctggaggaca gagtggcctc gagtcagacg 2701 tccctccgaa cgcaggcccc ggcgcagagg gcgggggcag ctggaagggg cggcctttcc 2761 cgtgcggcgc ctgtggccgc agcttcaagt gctcctcgga cgcggcaaag caccggagca 2821 tccactcggg ggagaaaccg tacgagtgca gcgactgcgg gaaggccttc atccacagct 2881 cgcacgtggt ccggcaccag cgggcgcaca gcggggagag gccctacgcg tgcgccgagt 2941 gcggcaaggc cttcggccag agcttcaacc tcctccggca ccagcgcgtg cacacgggcg 3001 agaagcccta cgcgtgcgcc gactgcggca aggccttcgg ccagaggtcg gacgccgcca 3061 agcaccgccg cacccacacc ggggagaggc tgtacgcgtg cggcgagtgc gggaagcgct 3121 tcctgcacag ctcgaacgtg gtccggcacc ggcggaccca ccacggggag aacccgtacg 3181 agtgccggga gtgcggccag gccttcagcc agagctccaa cctcctccag caccagcgcg 3241 tgcacacggg ggagcggccc ttcgcctgcc aggactgcgg ccgcgccttc agccgcagct 3301 ccttcctccg cgagcaccgc cgcatccaca ccggggagaa gccccaccag tgcggccact 3361 gcgggcgcgc gttccgggcg ctgtcgggct tcttccggca ccagcggctc cacacgggcg 3421 agaagccgtt ccgctgcacc gagtgcggcc gcgccttccg cctgagcttc cacctcatcc 3481 agcaccggcg ggtgcatggc gccgagggga ttccaactac tgcaagcgag aatctttatt 3541 ttcagggcgc cgccaaattc aaagaaaccg ctgctgctaa attcgaacgc cagcacatgg 3601 acagcggagg tggaggttcg agcggtccct cgggttcgtc gagcctggaa gttctgttcc 3661 aggggcccct gtcgtcgagc ggtccctcgg gttcgatggt gagcaaaggc gaggagctgt 3721 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca 3781 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct 3841 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg 3901 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca 3961 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga 4021 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca 4081 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc 4141 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc 4201 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca 4261 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga 4321 gcaaagaccc caacgagaag cgcgatcaca tggtccttaa ggagttcgtg accgccgccg 4381 ggatcactct cggcatggac gagctgtaca agtaatgatc tagagtcgag ttaattaaga 4441 attccgcccc cccccctctc cctccccccc cctaacgtta ctggccgaag ccgcttggaa 4501 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat 4561 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct 4621 ctcgccaaag gaatgcaggg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct 4681 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc 4741 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa 4801 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc 4861 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg 4921 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc 4981 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaacctg 5041 ttacattgca caagataaaa atatatcatc acgaacagta aaactgtctg cttacataaa 5101 cagtaataca aggggtgtta tgggatcggc cattgaacaa gatggattgc acgcaggttc 5161 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg 5221 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac 5281 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc 5341 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg 5401 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga 5461 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg 5521 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg 5581 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt 5641 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc 5701 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg 5761 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga 5821 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc 5881 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagccgggg ctgcggcgga 5941 agagatgccg gcggcctggt gggcgcgagg ccgaggccgg gggaggctcc tgtccgcccc 6001 gtgcgcggtg aggaagtaac agccggctgc gcgcctggtt ctcgggttgc gaaagcctcg 6061 ccaggcctgc atccgccttg gcttgggagc aatcaggaga gacagggctc ggggaaggcg 6121 agcgctgccc gcgggaggcg attcccagag gcggggaggt ctcaggggtc tgtcccgggc 6181 cggccgcccg cctctgagac tcccagcctc ccacggtcag ctgctgcggg gcagtggccg 6241 gtgattgaat cggtcattcc tccgcaacca ggcctccatc aaaaccctgc atgagatgag 6301 ggcctcgggc ttcgggttgg tgaaggcggg gtgggggggc agttaggaag ggggcgccca 6361 gacagggcat ggaagcctcg cgtcctcccc cgtaccttgc gggtcggtgg ttcatccatg 6421 tatttcaacg ccttttcaca gccggttcat atggtcccga cttctgtgtg cttagaaaat 6481 tatggaactt ggccgggtgc ggtggctccc gcctgtaatc ctagcacttt gggaggccga 6541 ggcgggcgga tcacacggtc aagagatcaa gaccatcctg gccaacatgg caaaaccccc 6601 atctcgacta aaaatacaaa aaattaattg ggcgtggtgg catgcgcctg taatctcagc 6661 tacttgggag cctggggcag gagaatcact tgaacctggg aggcggaggt tgcagtgagc 6721 ggagatctgg gcactgtact ttagcctggg cgacagagag acccatctc //