LOCUS Exported 7278 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor ZNF462 with GFP. ACCESSION . VERSION . KEYWORDS pZNF462-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7278) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 7278) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7278 /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 3872..3892 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3905..3949 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3989..4012 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 4040..4759 /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(4085..4106) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4346..4365) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4797..5382 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4963..4980) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5190..5209 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5464..6267 /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(5527..5546) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6137..6156 /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 tagaaggaca 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 gtcacccagc cgcaagtcaa atagggaaaa 2701 aaatcgtctt tatgaatgat cagtcctgct caagcctgtg tgcagaagta tagtgttttc 2761 tccagccaat atccctttag tcttcattag cgtgacattc tataatagaa cgtgaatcga 2821 ggtcaaaaga tctggcttct gttcctgcct ctctgactga tcattccctg agtggccctg 2881 aacattccca tgtactgcct cggggacaga gccagcttct gcctgcctcc caggtaggtg 2941 agagcatact tgtaaaatac ttatgagatc cctgaacata aaatcagccc ctcaacaagg 3001 cacgttagtc caccagatgc agagagaacc taggatgtgg tcttttcagg aggaaacatg 3061 gcttgtgtgg tgataggaag cctgaagctt ggtccctgaa gcctcaaggg tctagacctc 3121 agagaatata accaagcttc ccgagacatg ggttccatct tcaggttttt gtttttagtt 3181 tgttttcgtg atttgtttcg ggacctgaca aataaatctt tagtacctga gttactcttt 3241 tagacttcat aaatataacc aatctccaag cttctgggtt cagccgatga ctctgagcat 3301 aagacaattc ttgctgttac cacttatgac ccagtagaga aagtgacata aaatgattat 3361 gcttatctag aaagaaaaca tgggataatc aatatatagt ataataatgg atacttctgt 3421 aagttcaaat ctttaagata cagaaatata tgacttaagc aaaacatagc agggatgcat 3481 cacttggtat tttgtcatac acccatggca tttgttcaaa ggaaacccca aggctttttg 3541 aggatcagga aaataaagac actcgagggt ttttattatt tttttgttta aaaagagaaa 3601 tatatatact atactcatct gtctcttcga taaatgtttt tccagggcat tgaatgacac 3661 caagcaggtg agcagagaag aaatccaccc aaaagagatc atggagaaca gtgttaaaat 3721 gccctccata gaggaaaagg aagatgacga ggccattggg atagactttt ccctaaagaa 3781 tgaaacagta gccatctgtg tagtaactgc cgacaaatct ctcctggaga atgcagaggc 3841 caaaaaagaa gggattccaa ctactgcaag cgagaatctt tattttcagg gcgccgccaa 3901 attcaaagaa accgctgctg ctaaattcga acgccagcac atggacagcg gaggtggagg 3961 ttcgagcggt ccctcgggtt cgtcgagcct ggaagttctg ttccaggggc ccctgtcgtc 4021 gagcggtccc tcgggttcga tggtgagcaa aggcgaggag ctgttcaccg gggtggtgcc 4081 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg 4141 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct 4201 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg 4261 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt 4321 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa 4381 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga 4441 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat 4501 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga 4561 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt 4621 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga 4681 gaagcgcgat cacatggtcc ttaaggagtt cgtgaccgcc gccgggatca ctctcggcat 4741 ggacgagctg tacaagtaat gatctagagt cgagttaatt aagaattccg cccccccccc 4801 tctccctccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt 4861 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac 4921 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc 4981 agggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa 5041 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg 5101 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg 5161 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc 5221 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat 5281 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg 5341 tggttttcct ttgaaaaaca cgatgataat atggccacaa cctgttacat tgcacaagat 5401 aaaaatatat catcacgaac agtaaaactg tctgcttaca taaacagtaa tacaaggggt 5461 gttatgggat cggccattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg 5521 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg 5581 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc 5641 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct 5701 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa 5761 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg 5821 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa 5881 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat 5941 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg 6001 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc 6061 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac 6121 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg 6181 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc 6241 tatcgccttc ttgacgagtt cttctgagtc gcgtttggtg aaattcttaa tcaaacctta 6301 cttgaacagt gatgaaaaag tgggagggct ggcttgggct gagaagggag ggacagaaaa 6361 gagaagacag aacaaagctg ctttttagga ctgaacaatc tattttcaaa gcactggtac 6421 ctgtgtgagt gagtatgtaa attaaagtta tttaaatggt tggaatatgt ggctcctttt 6481 ccatcactac atcttttctt ccggatcttc atcatggaag tttcatttgt tgcggaatat 6541 ggaagcacct cccaatggta cggtgcaccc tgtggtggtc ttggacagta tgtggaaaca 6601 gaagctccat gacggtagaa gacttctcat tggggagcaa ctttttgacg cacaactttt 6661 ggtgcgtttt tctagtttta ataccttaag ctttttcaag acctaactgc agccgctttg 6721 ggaaaaaaaa acaaaaaaca aaaaacagaa aacaaaaaaa aaaaaactgc tttgcataaa 6781 acagtcacct gtttcctagt acctcaaact taaccaaggg aactctctgc atttgtcagt 6841 actacctgtc gtcgttgtgg ttaaatgtag agagaactgc tgggcaagat ggtgaatgga 6901 gaaaaaaaaa gagttttaaa gaaaggaaca caaattgtgc ttaaaatccc catgtgggtt 6961 ttatttctta aaatactgtg atttttttaa ttattttagt aaaaaacaaa acaaaaaaaa 7021 gaaacagact ttaattatta gataactgtt tgtgaattgt catcctttat tccaggtaag 7081 ctgtttatta gtgttcaact ataatttaga atttggtaga ttcatgtgag ctaattttaa 7141 ggtgattgtg gagttttgtt tgccacatgt ttattttcta tcaatttgag tgttacaaac 7201 acagcacaat tcagtttttc atatgaattg tttttcgtgt gtgtgttgtt attgttgttt 7261 taatttcggg gcaaggga //