LOCUS Exported 7035 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor ZNF565 with GFP. ACCESSION . VERSION . KEYWORDS pZNF565-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7035) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 7035) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7035 /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 3675..3695 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3708..3752 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3792..3815 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 3843..4562 /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(3888..3909) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4149..4168) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4600..5185 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4766..4783) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4993..5012 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5267..6070 /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(5330..5349) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5940..5959 /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 tcggtaccct gtgggaaagc atttagccgt ggctcacacc 2701 ttattcaaca tcagaaaatt cacactggtg aaaaaccctt tggatgtaag gaatgtggga 2761 aggccttcag ccgtgcctca caccttgttc agcatcagag aattcacacg ggtgagaaac 2821 cttatgactg taaggactgt gggaaggcct ttggtcgtac atcagaactt attctacatc 2881 agagacttca tactggtgtc aaaccttatg aatgtaaaga atgtgggaag acctttaggc 2941 agcattcaca gctgattctg catcaaagaa ctcacacagg cgagaaaccc tacgtatgta 3001 aagactgtgg caaggctttc attcgtggct cccaactcac tgtgcatcgg agaatccaca 3061 caggggccag accctatgag tgtaaagaat gcgggaaagc ctttagacag cactcacagc 3121 tgactgtaca ccagcgaatc cacactggtg agaaacccta cgagtgtaag gaatgcggaa 3181 agggctttat tcacagctca gaagttactc gacatcaaag aattcattct ggggagaaac 3241 cctatgagtg taaggaatgt gggaaggcct tcagacagca cgcacagctc acacgacatc 3301 agagagtcca tactggcgac agaccctatg aatgtaagga ctgcgggaag gcatttagtc 3361 gtagctcata cctcattcaa catcaaagaa ttcatacagg tgacaaaccc tacgaatgta 3421 aggaatgtgg gaaggccttt attcgtgttt cacaactgac tcatcatcag cgaattcaca 3481 cttgtgagaa accctatgaa tgcagggagt gtggaatggc ctttattcgt agttcacaac 3541 ttaccgaaca tcagagaatt catcctggta tcaaacctta cgaatgtaga gagtgtgggc 3601 aggcatttat tcttggctca caactcattg aacactacag aattcatact ggtgggattc 3661 caactactgc aagcgagaat ctttattttc agggcgccgc caaattcaaa gaaaccgctg 3721 ctgctaaatt cgaacgccag cacatggaca gcggaggtgg aggttcgagc ggtccctcgg 3781 gttcgtcgag cctggaagtt ctgttccagg ggcccctgtc gtcgagcggt ccctcgggtt 3841 cgatggtgag caaaggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg 3901 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct 3961 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca 4021 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga 4081 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct 4141 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc 4201 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc 4261 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga 4321 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg 4381 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc 4441 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg 4501 tccttaagga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt 4561 aatgatctag agtcgagtta attaagaatt ccgccccccc ccctctccct ccccccccct 4621 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt 4681 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg 4741 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcagggtct gttgaatgtc 4801 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt 4861 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta 4921 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg 4981 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag 5041 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag 5101 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa 5161 acacgatgat aatatggcca caacctgtta cattgcacaa gataaaaata tatcatcacg 5221 aacagtaaaa ctgtctgctt acataaacag taatacaagg ggtgttatgg gatcggccat 5281 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta 5341 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca 5401 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga 5461 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga 5521 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct 5581 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg 5641 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga 5701 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca 5761 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga 5821 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg 5881 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc 5941 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt 6001 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga 6061 gttcttctga gtctagaaag ccttgagtgt aagggataaa gataagcctt tatgtggagt 6121 acacatctgg ctcagtacta ggtaattttt aatgtagtgt aattaatgtc agaaaacacc 6181 tgtcacttcc atcatagaac atcagaaaac tcacacccaa agaaaattaa ataaatgtgg 6241 gaattctttt tgcctcactc actgtttcct aagcatcaga taatctatgc taaaaaaaaa 6301 tgaatataga aaagctgctt cttgccattc aaaatatgtt aaatttctga caattcctaa 6361 tacaaaaatg acccgattaa aagattctgt ggccaggcgt ggtggctcat gcctgtaatc 6421 ccagcacttt gggaggccaa ggccggcaga tcatttgagg tcaggagttt gagaccagcc 6481 tggccaatgt ggcgaaaccc catcactact aaaaatacaa aaattagcca ggcatggtgg 6541 cgggtgcctg taatcccagc tactcaggag gctgaggcag gaaaaccgtt tgaccccagg 6601 aggcagaggt tgcagcaagc cgagatccca ccattgcatt ccagcctcgt tgacaaggac 6661 aaaactctgt ctcaaaaaaa aaaaaaaaaa aaaaggttct ggctgggcat agtggctcac 6721 gcctgtaatc ccagcactct gggaggccga ggcgggcgga tcacaaggtc aggagatcga 6781 gaccatcctg gctaacacag tgaaacctgt ctcaactaaa aactatgaaa aaattagctg 6841 ggcctagtgg cgggcacctg tagtctcagc tactcgggag gctgaggcag gagaatggcg 6901 tgaacccagg aggcggagct tgcagtgagc caagatcgca ccactgcact ccagcctcgg 6961 cgacagagca agactccatc tcagaaaaaa aaaaaaaaaa gattctatga ctgtgccttt 7021 catcatggcg tatac //