LOCUS Exported 7124 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor ZNF639 with GFP. ACCESSION . VERSION . KEYWORDS pZNF639-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7124) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 7124) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7124 /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 3730..3750 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3763..3807 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3847..3870 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 3898..4617 /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(3943..3964) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4204..4223) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4655..5240 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4821..4838) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5048..5067 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5322..6125 /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(5385..5404) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5995..6014 /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 tcggtaccca attgctgtag aagtgcatgc gatttctgag 2701 gattatgata tagagacaga aaacaattcc tctgagagtc tccaagacca aactgatgaa 2761 gaaccgccag ctaaactttg taaaattctt gacaagagcc aagctttgaa tgtgactgcc 2821 cagcagaaat ggcctttact gagagctaat agcagtggcc tctataaatg tgaactttgt 2881 gagtttaaca gcaaatattt ttctgactta aagcagcata tgatcctgaa gcataaacgt 2941 actgattcaa atgtgtgtcg agtatgcaag gaaagtttct ctaccaatat gcttctgata 3001 gaacatgcca aactgcatga agaggatccc tacatttgta aatactgtga ttataagaca 3061 gtaatttttg agaacctcag ccagcacatt gcagacaccc attttagtga tcacctctat 3121 tggtgtgaac agtgtgatgt acagttctcc tcaagcagtg aactctacct acatttccag 3181 gagcacagct gtgatgaaca gtacttgtgt cagttctgtg aacatgaaac taatgatcca 3241 gaagacttgc atagccatgt ggtaaatgag catgcatgta aattaataga gttaagtgat 3301 aagtataaca atggtgaaca tggacagtat agcctcttaa gcaaaattac ctttgacaaa 3361 tgtaaaaact tctttgtatg tcaagtatgt ggttttcgga gtagacttca cacaaatgtt 3421 aacaggcatg ttgctattga acatacaaaa atttttcctc atgtttgtga tgactgtggg 3481 aaaggctttt caagtatgct agaatattgc aagcatttaa attcacattt atctgaaggg 3541 atttatttat gtcaatattg tgaatattca acaggacaaa ttgaagatct taaaattcat 3601 ctagatttca agcattcagc tgacttgcct cataaatgta gtgactgctt gatgaggttt 3661 ggaaatgaaa gggaattaat aagtcacctt ccagtccatg agacaactgg gattccaact 3721 actgcaagcg agaatcttta ttttcagggc gccgccaaat tcaaagaaac cgctgctgct 3781 aaattcgaac gccagcacat ggacagcgga ggtggaggtt cgagcggtcc ctcgggttcg 3841 tcgagcctgg aagttctgtt ccaggggccc ctgtcgtcga gcggtccctc gggttcgatg 3901 gtgagcaaag gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc 3961 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc 4021 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc 4081 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag 4141 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc 4201 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg 4261 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag 4321 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc 4381 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac 4441 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac 4501 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctt 4561 aaggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatga 4621 tctagagtcg agttaattaa gaattccgcc cccccccctc tccctccccc cccctaacgt 4681 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac 4741 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag 4801 cattcctagg ggtctttccc ctctcgccaa aggaatgcag ggtctgttga atgtcgtgaa 4861 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag 4921 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga 4981 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag 5041 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc 5101 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag 5161 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg 5221 atgataatat ggccacaacc tgttacattg cacaagataa aaatatatca tcacgaacag 5281 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgggatcg gccattgaac 5341 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact 5401 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc 5461 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg 5521 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg 5581 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt 5641 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc 5701 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag 5761 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg 5821 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc 5881 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt 5941 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg 6001 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt 6061 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct 6121 tctgagtctg attattctct ttaacttaca gaatgttagt ttaaaataat aaattcatcc 6181 tttttttgga gatgattaaa tggatgattg taaacacaac ttatgaaatc tgcctttaac 6241 aagtaacttt tttaaattat aaaattttat tggcattgct ccattttctg tatataaata 6301 tatctttaat gtggtatttt caattgcgtg atagtttgta gtttcaacca ctcttggtga 6361 ctgtcatcct gtttcttcca tattctctga tttcatgaat tgaaaagaaa caaatgtatt 6421 gaagaagtga gctacagttt tccttcctta accatgggtg ctagtaactt tttaaaactc 6481 aagacaagat tagtttttta tgtgtgaagt cattaaatta ttacacgacc agaactaaaa 6541 tgcaatatac agttaagtcc acggatactc ccattaatga gaaataacac taggaagcca 6601 ctattacagg aagaaaagat ttggttttca tggcagtctg tttttttaaa aaaaaatttt 6661 tgagccacta tctattgttg aatattttaa gatgggatga gggaggaact aataagggct 6721 tacacaataa aaaataacta tatcataact cattcataac ttgatgtttc attttctgtt 6781 gaggaaccat aaattcattc acagacttaa tatttttttc ttagagatgg tctcgttctg 6841 ttgtccagga tggagtgcag tggttgatca tagctcccgg gctgtagtct cccaggctcg 6901 agcaatcctc ccacttcacc ctcctgcata gctaggacta caggcatgtg gcaccatgcc 6961 cgctaagttt ttaaatttct tgtagagatg agatctcgct atgttgccca ggccagtctc 7021 aaactcctgg actcaagcaa tcctcccacc ttggccttcc aaatcactgg gattataggc 7081 atgagccatt atgcctgact cttgcccaaa tttctgatgt caaa //