LOCUS Exported 7068 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor ZNF695 with GFP. ACCESSION . VERSION . KEYWORDS pZNF695-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7068) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 7068) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7068 /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 3623..3643 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3656..3700 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3740..3763 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 3791..4510 /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(3836..3857) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4097..4116) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4548..5133 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4714..4731) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" CDS 5215..6018 /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(5278..5297) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5888..5907 /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 tcggtaccct ccatattgga gagaacccgt accaatgtaa 2701 aaaatgtggc aaagccttta atgagtgctc atgctttact gactgtaaga gaattcatgt 2761 tggagagaaa cattgcaaat gtgaagaatg taataacatt tttaagtctt gctcaagtct 2821 tgctgttgtt gagaaaaatc atactgaaaa gaaaacctac agatgtgaag aatgtggcaa 2881 agcttttaac ctgtgctcag ttcttactaa acataagaaa attcatactg gagagaaacc 2941 atacaaatgt gaagaatgtg gcaaatcctt taagttgttc ccatacctta ctcaacacaa 3001 gagaattcat agtagagaga aaccctacaa gtgtgaagaa tgtggcaaag tctttaaatt 3061 gttgtcatac cttactcaac atagaagaat tcatactgga gagaaaacct tccgatgtga 3121 agaatgtgga aaagccttta accagagctc acatctgact gaacatagga gaattcatac 3181 tggtgagaaa ccatacaaat gtgaggaatg tggcaaagct tttacctggt tctcatacct 3241 tattcagcat aagagaattc atactgggca gaaaccctac aaatgtgagg aatgtggcaa 3301 agcttttacc tggttttcat accttactca acataagaga attcatactg gagagaaacc 3361 ctacaaatgt gatgaatgtg gcaaagcttt taactggttt tcatatctta ctaatcataa 3421 gagaattcat actggagaga aaccctacaa atgtgaagaa tgtggcaaag cctttggcca 3481 gagctcacac ctttctaaac ataagacaat tcataccaga gagaaaccat acaagtgtga 3541 ggaatgtggc aaagccttta accactctgc acaacttgct gtacatgaga aaactcatac 3601 cgggattcca actactgcaa gcgagaatct ttattttcag ggcgccgcca aattcaaaga 3661 aaccgctgct gctaaattcg aacgccagca catggacagc ggaggtggag gttcgagcgg 3721 tccctcgggt tcgtcgagcc tggaagttct gttccagggg cccctgtcgt cgagcggtcc 3781 ctcgggttcg atggtgagca aaggcgagga gctgttcacc ggggtggtgc ccatcctggt 3841 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga 3901 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc 3961 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga 4021 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg 4081 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg 4141 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat 4201 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa 4261 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt 4321 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc 4381 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga 4441 tcacatggtc cttaaggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct 4501 gtacaagtaa tgatctagag tcgagttaat taagaattcc gccccccccc ctctccctcc 4561 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata 4621 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg 4681 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg cagggtctgt 4741 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag 4801 cgaccctttg caggcagcgg aaccccccac ctggcgacag gttcctctgc ggccaaaagc 4861 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga 4921 tagttgtgga aagagtcaaa tgtctctcct caagcgtatt caacaagggt ctgaaggatg 4981 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat 5041 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc 5101 tttgaaaaac acgatgataa tatggccaca acctgttaca ttgcacaaga taaaaatata 5161 tcatcacgaa cagtaaaact gtctgcttac ataaacagta atacaagggg tgttatggga 5221 tcggccattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 5281 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 5341 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 5401 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 5461 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 5521 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 5581 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 5641 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 5701 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 5761 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 5821 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 5881 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 5941 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 6001 cttgacgagt tcttctgagt ctgagaaaaa ccctacaaat ctaaacaata tggcatagtc 6061 tttaatacct attcacaact tcacagcaga atatttttac tgaataagag tgttacaaat 6121 gtaatgactg tcaaaaggcc atttacagtc tatgagcctt tgagtgcact aaaatgttta 6181 ggctacgaac aatacaaata gaccggttca acacctccac ttatatcaca gctcttactg 6241 tacacagaag aatttatact ggagggaaac cctccagttg ctcaaactgt attcaatttc 6301 aaagactttg tattggagag aaaccctaca aatgtaataa atgcagaaac aacatttgtt 6361 caaaaaatat acctcagaaa acaccagagt gttcacacta aaaactgttt tacagatgca 6421 gtaaatgtga aaaaatgtct aatcaaaaat tacatcaaaa cacatccaag aattcatagt 6481 aaaaagcact aagtcactga cactttcaga cattactgta aatctgagtg ttggttatag 6541 agaataattc aaagttaagt taaagtaagt aggagattca ccttttgggg aagttataat 6601 tacatttcaa gtataccttt tggtgccagg cacggtggct ctttcctata gttgctgcac 6661 ttttggatgc cgaggtcggg ggattgcttg agcccaggag tttgggacca ggctgggcaa 6721 catggcaaaa cctcatctct acaaaaagta aaataaaagc caggcttggt ggcacatgcc 6781 tgttgtccca gctacttgga aggctcaggt gggaggattg cttgagcctg gaggttgagg 6841 atgcagtgag ctgctatctg gcaacttcac tcctgcctgg gcaacagagc aagacccttt 6901 ctcaataata ataataacaa cataataata aagtatactc ggtgcactga aagagtttta 6961 gcttttttga aaatcacata tttatgtaat tcaagtctta aatcacttga taccatgcct 7021 tcatttctag tgtttatgtg aaggcatgag gcctactgtt gctacatg //