LOCUS Exported 6850 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor ZNF700 with GFP. ACCESSION . VERSION . KEYWORDS pZNF700-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6850) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 6850) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6850 /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 3558..3578 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3591..3635 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3675..3698 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 3726..4445 /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(3771..3792) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4032..4051) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4483..5068 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4649..4666) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4876..4895 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5150..5953 /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(5213..5232) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5823..5842 /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 tccttctagt 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 ctttaaggaa tgtgggaaag ccttcagatc tacctcacac cttcgagtgc 2701 atggtaggac tcatactgga gagaaaccct atgaatgtaa ggaatgtggg aaagccttca 2761 gatatgtgaa gcaccttcaa attcatgaaa ggacagaaaa acacataaga atgccctctg 2821 gagaaagacc ttataaatgt agtatatgtg agaaaggctt ttattctgcc aagtcatttc 2881 aaacacatga aaaaactcac actggagaga aaccctatga atgcaaccaa tgtggtaaag 2941 ccttcagatg ttgcaattcc cttcgatatc atgaaaggac tcacactgga gagaaaccct 3001 atgagtgtaa gcaatgtggg aaagccttca gatctgcctc acaccttcga atgcatgaaa 3061 ggactcacac tggagagaaa ccctatgagt gtaagcaatg tgggaaagcc ttcagttgtg 3121 cctcaaacct tcgaaagcat ggtaggactc acactggaga gaaaccctat gagtgtaagc 3181 aatgtgggaa agccttcaga tctgcctcaa accttcagat gcatgaaagg actcacactg 3241 gagagaaacc ctatgaatgt aaggaatgcg aaaaagcatt ctgtaaattc tcttcttttc 3301 aaatacatga aaggaagcac agaggagaga agccctatga atgtaagcat tgtgggaatg 3361 gattcacatc tgccaagatt cttcaaatac atgcaagaac acacattgga gagaaacact 3421 atgaatgtaa ggaatgcgga aaagcattca attatttttc ttccttgcat atacacgcaa 3481 ggactcatat gggagagaag ccatatgaat gtaaggattg tgggaaagca ttcagcggga 3541 ttccaactac tgcaagcgag aatctttatt ttcagggcgc cgccaaattc aaagaaaccg 3601 ctgctgctaa attcgaacgc cagcacatgg acagcggagg tggaggttcg agcggtccct 3661 cgggttcgtc gagcctggaa gttctgttcc aggggcccct gtcgtcgagc ggtccctcgg 3721 gttcgatggt gagcaaaggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc 3781 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca 3841 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc 3901 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca 3961 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca 4021 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca 4081 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg 4141 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga 4201 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc 4261 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca 4321 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca 4381 tggtccttaa ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca 4441 agtaatgatc tagagtcgag ttaattaaga attccgcccc cccccctctc cctccccccc 4501 cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta 4561 ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc 4621 ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaggg tctgttgaat 4681 gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc 4741 ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt 4801 gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt 4861 gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag 4921 aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt 4981 tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga 5041 aaaacacgat gataatatgg ccacaacctg ttacattgca caagataaaa atatatcatc 5101 acgaacagta aaactgtctg cttacataaa cagtaataca aggggtgtta tgggatcggc 5161 cattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg 5221 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc 5281 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca 5341 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct 5401 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga 5461 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg 5521 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat 5581 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga 5641 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg 5701 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg 5761 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat 5821 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct 5881 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga 5941 cgagttcttc tgacctggtt ccttttatgg acatgaatag actcacactg gaaggaagca 6001 ctatgaatgc aagcaatgtg gcaaaacttt cacattttcc agttcttttc gatatcatga 6061 aaggactcac actggggaga aaccctatca atgtaagcag tgtgggaaag ccttcattcc 6121 ttttacttct tttcaatgtc atgaaaggac tcacacggga gagaaaccct atgagtgtat 6181 tctagttccg tttgatatca tgaaaggact tacactggag tgaaacccta tgaatgtaag 6241 caatgtggga aagccttcag atgtgcctcg caccttcaac ggcatggaag ggttcacact 6301 tgggagaaac tctatgaatg taagcagtat gggaaagcct tcagatctgc caagattctt 6361 tgaatacaga taattaatgt aaacaattat cataagtata ctaacatgtt attcttttta 6421 aataagaagg tataataaaa tatcccattg gttttatgta ttagatcaag cttataatgt 6481 tacattgtta ttatttggac attgtgagtc agtattacca tgtggataaa atgccaggca 6541 tctttttcct catggaaagt ttactttttg caccgggcac agtggctcac gcctgtaatc 6601 ccagcacttt gggaggccaa ggcaggcgga tcacgaggtg aagagatcaa gaccattctg 6661 accaacgtgg tgaaaccctg tctctactaa aaatacaaaa attagctggg catggtggcg 6721 cgcaccggta gtcccagctt ctcgggaggc tgaggcagga gaatcgcttg aacctgggag 6781 gcggaggttg cagtgagcag agatcacacc actgcactcc agcctggcaa cagagcaaga 6841 ctccatctcc //