LOCUS Exported 7377 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor ZNF416 with GFP. ACCESSION . VERSION . KEYWORDS pZNF416-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7377) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 7377) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7377 /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 3888..3908 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3921..3965 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 4005..4028 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 4056..4775 /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(4101..4122) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4362..4381) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4813..5398 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4979..4996) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5206..5225 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5480..6283 /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(5543..5562) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6153..6172 /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 tcggtacccg agaagccaaa caaaatcagc aaatgtgagg 2701 aggcctttca tgttggaata agtcattaca agtggagtca atgcaggaga gagtccagcc 2761 acaaacacac tttttttcac cctagagtct gcactggaaa aaggctttat gaatctagca 2821 aatgtgggaa agcctgctgc tgtgagtgct cccttgttca gctgcaaaga gtccaccctg 2881 gagaaaggcc ttatgagtgc agtgaatgtg ggaaatcttt tagccaaacc tctcatctga 2941 atgatcatcg gagaatccac actggagaaa ggccttatgt gtgtggtcag tgtgggaaat 3001 catttagcca aagagccacc ctcattaaac atcacagagt tcacactgga gaaaggcctt 3061 acgagtgtgg tgaatgtggg aaatctttta gccaaagttc caaccttatt gaacattgca 3121 gaattcacac tggagaaagg ccttatgagt gtgatgaatg tggaaaagcc tttgggtcca 3181 aatccactct tgttcgacac cagagaactc acacaggaga aaagccatat gagtgtggtg 3241 aatgtgggaa attattcaga caaagcttca gccttgttgt acaccagaga attcacacta 3301 cagcaaggcc ttatgagtgt ggccagtgtg ggaaatcatt tagcctaaag tgtggcctca 3361 ttcagcacca gttaattcac agtggagcta ggccctttga gtgtgatgag tgcggaaaat 3421 cctttagcca aagaaccacc ctcaataaac accacaaagt tcacactgca gaaaggcctt 3481 atgtatgtgg ggaatgtggg aaagctttta tgttcaaatc taaacttgtt aggcaccaga 3541 gaactcacac tggagaaagg ccttttgagt gcagtgaatg tgggaaattt tttagacaaa 3601 gctataccct cgttgaacac cagaaaattc acactggatt aaggccttac gactgtggac 3661 agtgcgggaa atcctttatc caaaagtcta gcctcattca acaccaagtg gttcacacag 3721 gagaaaggcc atatgagtgt ggcaaatgtg ggaagtcctt tacacaacac tctggcctca 3781 ttctccaccg aaaatctcac actgtggaga ggcctcgtga cagcagcaaa tgtggaaaac 3841 cctacagccc aagatctaac attgttggga ttccaactac tgcaagcgag aatctttatt 3901 ttcagggcgc cgccaaattc aaagaaaccg ctgctgctaa attcgaacgc cagcacatgg 3961 acagcggagg tggaggttcg agcggtccct cgggttcgtc gagcctggaa gttctgttcc 4021 aggggcccct gtcgtcgagc ggtccctcgg gttcgatggt gagcaaaggc gaggagctgt 4081 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca 4141 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct 4201 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg 4261 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca 4321 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga 4381 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca 4441 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc 4501 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc 4561 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca 4621 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga 4681 gcaaagaccc caacgagaag cgcgatcaca tggtccttaa ggagttcgtg accgccgccg 4741 ggatcactct cggcatggac gagctgtaca agtaatgatc tagagtcgag ttaattaaga 4801 attccgcccc cccccctctc cctccccccc cctaacgtta ctggccgaag ccgcttggaa 4861 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat 4921 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct 4981 ctcgccaaag gaatgcaggg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct 5041 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc 5101 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa 5161 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc 5221 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg 5281 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc 5341 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaacctg 5401 ttacattgca caagataaaa atatatcatc acgaacagta aaactgtctg cttacataaa 5461 cagtaataca aggggtgtta tgggatcggc cattgaacaa gatggattgc acgcaggttc 5521 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg 5581 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac 5641 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc 5701 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg 5761 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga 5821 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg 5881 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg 5941 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt 6001 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc 6061 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg 6121 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga 6181 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc 6241 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagtcctct tgaaactcca 6301 aacctgagaa aagccttaga cctgcaggga atgtgccatg tctttcttca gtgttatggc 6361 tatagagagc ctttgtgaag gagccgtctg cctaaagtta aacctcatac ttctagacct 6421 cccgatgggt cagactccca ttagttccag gcatatgtgt cttgcaggag ctcctttgca 6481 tgttctaaac tgctggaggc ctttgccaga gttaaatcat tagtcaccct tgcatactaa 6541 ggcgaggcag atacacctgt gttctcccaa cccttggagg aaattccttg agcagtctga 6601 gcctttaagg ggaattcctt cccttttctg actgattaca gcatacatct gaccagtttt 6661 ggccagaagg gcccagcctg ggttctgctg gaagctgatg tggaaggctt cccttcatgg 6721 aaattcttgt tttcacacaa cacttggtag tttttccagc ctccaagtca ccacctagta 6781 actggctgcc tcacattgct ctggtttgtg tggtaataaa ggccttattt ttaaatctat 6841 gtgtggtctt agggtttggt ttactgtgtg aggtggctgg aaaacacatt tgggctccta 6901 cataaaggca tggacagctt ttgtggggcc ccaatgttgc tggcctgtgc aggaaaagta 6961 aaatgacagt agttcttgtt ctggttttag tccactttgc cttgctgctc aaggcctcct 7021 aggaaacaat gcaagaattt tgttgtccta atttatggcc tggatcccta cactttctct 7081 ggaccagagg tcatcctgag gacaagtcgg ctgttatgac aggctcctgt gctttaggga 7141 ataagatgga tttattgcat tgttctacag gctgtaatgc ccagcactgt tgagacgaag 7201 ctttttccca gcttctgcaa aggaacctat gtgccctgat gcccatgatt ccatagatgg 7261 tggtactcaa gctgatggga gctaagcatg ggacataatt ggtacagaaa ttttgggtat 7321 gattaataga aagtggagga ggtaatagca aaacaaatgt gcctgtggta aaagtgc //