LOCUS Exported 6790 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor ZNF23 with GFP. ACCESSION . VERSION . KEYWORDS pZNF23-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6790) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 6790) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6790 /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 3575..3595 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3608..3652 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3692..3715 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 3743..4462 /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(3788..3809) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4049..4068) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4500..5085 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4666..4683) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4893..4912 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5167..5970 /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(5230..5249) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5840..5859 /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 cttggcttca ggtgcagctc ccagcttagg cagcatcaga gcatccacac 2701 aggagaaaag ccctatcagt gtaaagagtg tggaaaaggc ttcaataata atacaaaact 2761 cattcagcat cagagaatcc acacaggtga gaaaccctat gaatgcactg aatgtggaaa 2821 agccttcagt gtcaaaggga agttaatcca acaccagaga attcacacag gcgagaaacc 2881 ctatgagtgt aatgaatgcg ggaaagcctt cagatgtaac tcccaatttc ggcagcatct 2941 gagaattcac actggggaga agccctatga gtgtaatgag tgtggaaagg ccttcagcgt 3001 taatgggaaa ctaatgcggc atcagagaat tcacactggg gagaaacctt ttgaatgtaa 3061 tgagtgtggg agatgcttta cttctaaaag aaacctactt gatcatcacc gaatccatac 3121 tggagaaaag ccctatcaat gtaaggaatg tgggaaagcc ttcagtatca atgccaaact 3181 aactaggcat cagaggatac atactgggga gaaacctttc aaatgtatgg aatgtgagaa 3241 agcattcagc tgtagttcta actatattgt gcaccagaga atccatacag gagagaaacc 3301 ctttcagtgt aaggagtgtg gaaaagcctt ccatgttaat gcccatttaa ttcggcatca 3361 gagaagccac actggggaga aacccttcag atgtgtggaa tgtggcaaag gcttcagctt 3421 tagttctgac tacattatac atcagacagt ccacacttgg aagaaaccct atatgtgtag 3481 tgtgtgtggg aaagcattca ggtttagctt ccagctcagt cagcatcaga gtgtccatag 3541 tgaaggaaaa tccgggattc caactactgc aagcgagaat ctttattttc agggcgccgc 3601 caaattcaaa gaaaccgctg ctgctaaatt cgaacgccag cacatggaca gcggaggtgg 3661 aggttcgagc ggtccctcgg gttcgtcgag cctggaagtt ctgttccagg ggcccctgtc 3721 gtcgagcggt ccctcgggtt cgatggtgag caaaggcgag gagctgttca ccggggtggt 3781 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga 3841 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa 3901 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag 3961 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta 4021 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt 4081 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 4141 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat 4201 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga 4261 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 4321 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa 4381 cgagaagcgc gatcacatgg tccttaagga gttcgtgacc gccgccggga tcactctcgg 4441 catggacgag ctgtacaagt aatgatctag agtcgagtta attaagaatt ccgccccccc 4501 ccctctccct ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg 4561 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga 4621 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa 4681 tgcagggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa 4741 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct 4801 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg 4861 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg 4921 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca 4981 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg 5041 acgtggtttt cctttgaaaa acacgatgat aatatggcca caacctgtta cattgcacaa 5101 gataaaaata tatcatcacg aacagtaaaa ctgtctgctt acataaacag taatacaagg 5161 ggtgttatgg gatcggccat tgaacaagat ggattgcacg caggttctcc ggccgcttgg 5221 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc 5281 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt 5341 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt 5401 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc 5461 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc 5521 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac 5581 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag 5641 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag 5701 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat 5761 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg 5821 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa 5881 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc 5941 ttctatcgcc ttcttgacga gttcttctga taatgagaaa gatatagaaa actcttaagg 6001 ttaatgccaa aatggatcaa gtatcatcag attcatccat tgaaaaacct ccaagagggc 6061 atgaatatgg cagagtcttc atatggaaac agtttttatt ctattcagtt taaatcagga 6121 aaggatgacc agttaaagag aaacatccaa aaatagcttt gttttgtacc aacaggaatt 6181 agaaaatata atgaaaagat ttcgttccca gcagcatcaa gaaaagtaga ttttctagaa 6241 ataaacagtt atggaggact tgtatggaga aatttaagtc ttcactgagg gccactttac 6301 aaaggaaatt tgaataaatg gagagagaga gaagccttgt tgttggatag gaaaacccgt 6361 actaaagata ctctacctac attaatttat ttgtttaatt tttgacaaca agcatgtatt 6421 acttttgaaa agatgaaaaa taaagtttta tttaaaaaag gaaaaaggta tgagtacatt 6481 tttaattttc atgtaattga gaaaaccttt ctaagtatta gatgaaagct agaaactatc 6541 cacagggtat gtactgataa attgactact caaaaactaa atattcctgt atgtttttgt 6601 tttaaagttt aaaagataaa gcgcaaaatg gggaaatact tttgtaaaat ttgtgatata 6661 aaagtgtagt atttgggcat ggtggatcag acctgtaatc tcagtgcttt gggaggctga 6721 gacgggaaga ttgcgtgagg ccatgagttc cagacctgta tgggcaacat agcaagatcc 6781 catcgctaca //