LOCUS Exported 6820 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor ZNF436 with GFP. ACCESSION . VERSION . KEYWORDS pZNF436-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6820) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 6820) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6820 /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 3566..3586 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3599..3643 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3683..3706 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 3734..4453 /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(3779..3800) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4040..4059) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4491..5076 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4657..4674) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4884..4903 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5158..5961 /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(5221..5240) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5831..5850 /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 gtcgcagctc acaccttatt cagcatcaaa gaacacatac 2701 tggggagagg ccttatgact gtaacgagtg tgggaaaagt tttggaagaa gttctcacct 2761 gattcagcat cagacaatcc acactggaga gaagcctcac aaatgtaatg agtgtggaaa 2821 aagtttctgc cgtctctctc acctaatcca acaccaaagg acccacagtg gtgagaaacc 2881 ctatgagtgt gaggagtgtg ggaaaagctt cagccggagc tctcacctag ctcagcacca 2941 gaggacccac acgggtgaga aaccttatga atgtaacgaa tgtggccgag gcttcagtga 3001 gagatctgat ctcatcaaac actatcgagt ccacacaggg gagaggccct acaagtgtga 3061 tgagtgtggg aagaacttca gtcagaactc cgaccttgtg cgtcatcgca gagcccacac 3121 gggagagaag ccataccact gtaacgaatg tggggaaaat ttcagccgca tctcacactt 3181 ggttcagcac cagagaactc acactggaga gaagccatat gaatgcaatg cttgtgggaa 3241 aagcttcagc cggagctctc atctcatcac acaccagaaa attcacactg gagagaagcc 3301 ttatgagtgt aatgagtgtt ggcgaagctt tggtgaaagg tcagatctaa ttaaacatca 3361 gagaacccac acaggggaga agccctacga gtgtgtgcag tgtgggaaag gtttcaccca 3421 gagctccaac ctcatcacac atcaaagagt tcacacggga gagaaacctt atgaatgtac 3481 cgaatgtgag aagagtttca gcaggagctc agctcttatt aaacataaga gagttcatac 3541 ggacgggatt ccaactactg caagcgagaa tctttatttt cagggcgccg ccaaattcaa 3601 agaaaccgct gctgctaaat tcgaacgcca gcacatggac agcggaggtg gaggttcgag 3661 cggtccctcg ggttcgtcga gcctggaagt tctgttccag gggcccctgt cgtcgagcgg 3721 tccctcgggt tcgatggtga gcaaaggcga ggagctgttc accggggtgg tgcccatcct 3781 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg 3841 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt 3901 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc 3961 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga 4021 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga 4081 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa 4141 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga 4201 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag 4261 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct 4321 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg 4381 cgatcacatg gtccttaagg agttcgtgac cgccgccggg atcactctcg gcatggacga 4441 gctgtacaag taatgatcta gagtcgagtt aattaagaat tccgcccccc cccctctccc 4501 tccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct 4561 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc 4621 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcagggtc 4681 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg 4741 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa 4801 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt 4861 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg 4921 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta 4981 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt 5041 tcctttgaaa aacacgatga taatatggcc acaacctgtt acattgcaca agataaaaat 5101 atatcatcac gaacagtaaa actgtctgct tacataaaca gtaatacaag gggtgttatg 5161 ggatcggcca ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg 5221 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg 5281 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat 5341 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca 5401 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg 5461 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat 5521 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa 5581 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg 5641 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg 5701 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg 5761 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat 5821 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac 5881 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc 5941 cttcttgacg agttcttctg agctgtaatt atgatggctg agaaatgatt catttgaaga 6001 tacaatttta tttgatatca atgaacgccc tcaagactga gctgctttta tcatactctc 6061 ctagttgtgg gccacgattt aaaccatcag agatgacaag ccatttgaaa ttctgaccct 6121 cagctttggg aatgttatct cctccaaaat ggtgattttt attcactcaa tgggttactt 6181 cattaaaagc agccccacaa gtaactggaa atctgaagac caggggacaa atgctggtga 6241 atgcctaggc ctggaaatgg agtaaatctt tcaatgttat tttctcccat ccttggccca 6301 aggaactatg ctaagtgaaa cgtgggactg taatagggtg gtaatggctg ctttggaaaa 6361 aggcaactag agactctgcc taaattgcca cacctattca cacaccatag tagttgggca 6421 cacacatctt cccttccaaa gggctttttc cttgagttgc tcatgcattt gtatcttttc 6481 catcttcctg agggcaagat tttgcacgat gaaggcaatg attgtaactt ttctccttct 6541 cattgtttct aattagctcc tttaaagctt gcatctttgt gaaggctaac tgaagatacg 6601 gttggaaagg aaaaatgaga cacaggtttg gggaccaagg acccatcaat gatggtgact 6661 ttagcagaag atgcccacag ttattactgc cattaatcag atttatgaat tttctttggg 6721 gatcactata gggaatattg tatagaaaat atcttcaaga aaagatagga ccatcagtga 6781 cagttaagtg taaggagcaa gtggaattga gtccttcagg //