LOCUS Exported 6963 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor ZNF192 with GFP. ACCESSION . VERSION . KEYWORDS pZNF192-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6963) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 6963) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6963 /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 3645..3665 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3678..3722 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3762..3785 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 3813..4532 /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(3858..3879) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4119..4138) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4570..5155 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4736..4753) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4963..4982 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5237..6040 /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(5300..5319) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5910..5929 /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 agaccaggag tgagaacagg gaattagctt 2701 caaaacaggt aatatctact ggaatccagc cacatggaga gacagctgcc aaatgcaacg 2761 gggatgttat caggggtctt gagcatgaag aagcccgaga ccttctgggc agattagaga 2821 ggcagcgggg aaatcccaca caagagagac gacataaatg tgatgaatgt gggaaaagct 2881 ttgctcagag ctcaggcctt gttcgccact ggagaatcca cactggggag aaaccctatc 2941 agtgtaatgt gtgtggtaaa gccttcagtt acaggtcagc ccttctttca catcaggata 3001 tccacaacaa agtaaaacgc tatcactgta aggagtgtgg caaagccttc agtcagaaca 3061 caggcctgat tctgcaccag agaatccaca ctggggagaa gccatatcag tgcaatcagt 3121 gtgggaaggc tttcagtcag agtgcgggcc ttattctgca ccagagaatc cacagtggag 3181 agagacccta tgaatgtaat gagtgtggga aagctttcag tcatagctca cacctcattg 3241 gacatcagag aatccacact ggggagaagc cctatgagtg tgatgagtgt gggaaaacct 3301 tcaggcggag ctcacatctt attggtcatc agaggagcca cactggggag aaaccctaca 3361 aatgcaatga gtgtgggagg gccttcagtc agaagtcagg ccttattgaa catcagagaa 3421 tccacactgg agaaagaccc tataaatgta aagaatgtgg gaaagctttc aatgggaaca 3481 ctggtctcat tcaacacctg agaattcaca caggggagaa gccctaccaa tgtaatgagt 3541 gtgggaaagc ctttattcag aggtcaagtc tcattcgaca tcagagaatc cacagtggtg 3601 aaaaatctga atccataagc gttgggattc caactactgc aagcgagaat ctttattttc 3661 agggcgccgc caaattcaaa gaaaccgctg ctgctaaatt cgaacgccag cacatggaca 3721 gcggaggtgg aggttcgagc ggtccctcgg gttcgtcgag cctggaagtt ctgttccagg 3781 ggcccctgtc gtcgagcggt ccctcgggtt cgatggtgag caaaggcgag gagctgttca 3841 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg 3901 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca 3961 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc 4021 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc 4081 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc 4141 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg 4201 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca 4261 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc 4321 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg 4381 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca 4441 aagaccccaa cgagaagcgc gatcacatgg tccttaagga gttcgtgacc gccgccggga 4501 tcactctcgg catggacgag ctgtacaagt aatgatctag agtcgagtta attaagaatt 4561 ccgccccccc ccctctccct ccccccccct aacgttactg gccgaagccg cttggaataa 4621 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg 4681 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc 4741 gccaaaggaa tgcagggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct 4801 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac 4861 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc 4921 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta 4981 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg 5041 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg 5101 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca caacctgtta 5161 cattgcacaa gataaaaata tatcatcacg aacagtaaaa ctgtctgctt acataaacag 5221 taatacaagg ggtgttatgg gatcggccat tgaacaagat ggattgcacg caggttctcc 5281 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc 5341 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga 5401 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac 5461 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct 5521 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa 5581 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc 5641 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct 5701 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc 5761 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg 5821 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct 5881 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct 5941 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca 6001 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gtctaggaac aacatcagtt 6061 agagtttgag cattattcag cattagggaa accacacact ggtgagaggt ctttcagtgt 6121 actaaaaggc agaaaggtca tcacaacttt agtgtcagaa tctatagtgg tggcaaagtt 6181 aggatagctc tttagtaatt tgggcccagt gccttggtga aggttgatta gcttgactgt 6241 ctttgaaagt atctgatgga gctcctgatg acctaatgta tcctttagaa atttaaaata 6301 gcattagagc aagttgcttg tcatggcttg aagagcttaa ctttgtcttt tgggtgagta 6361 gctataggtt tgagagaggc tggctactcc tagttcctgt gcttcttccc atctcctgtg 6421 atccccctct cccatttatt cccttgaagg tgcccttgta ttcctaatct gatctaagaa 6481 gctgctgtag agtcagaagt agcttctgtg tgaaacttat atttgtatgt agctttctag 6541 gaaaatttta cagacactta aagtatttat gcttaagtgt gcattttttt attctaatat 6601 tccttctagt cagtggaaat ttgtattccc atacaaactc agaatgctat atttttaaca 6661 gcactccagc atttttcttc atgtatcacc ttcactgact ccattgagtc attgagcatc 6721 tgtacatatc cttcaccacc cccagtccca gcacctgtgt cagtaaagaa agtggacaca 6781 ttagtaatgg ttatgggagt aatacagaga aagaggtgag tggagactca gcccagctgc 6841 tcttgagctt ggatagttat acattttggt gacttctgac tctccttcca cctccccact 6901 gtacactgtg gtacagatag ttggcactat cctagttatg gcatcaggta ggagaaggag 6961 acg //