LOCUS Exported 7372 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor ZNF215 with GFP. ACCESSION . VERSION . KEYWORDS pZNF215-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7372) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 7372) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7372 /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 3883..3903 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3916..3960 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 4000..4023 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 4051..4770 /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(4096..4117) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4357..4376) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4808..5393 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4974..4991) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5201..5220 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5475..6278 /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(5538..5557) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6148..6167 /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 gctagaagaa ggaatattag gcagagactt 2701 catacactag ttacgaacca gtgataactt gaattatgat atgatatgcc acccgcatcc 2761 ttgttcatgt aaagatattt tactcgagcc ctataccgtt tttttagtat tccagaatag 2821 ctctcctatt ttatttacta acctcctttt aacatttttg taccatttat ttcttttata 2881 agtatttttt tctaaccttg cctcacattt ccattctgta tttaagaccc gtggtatatg 2941 catatacctt atacagttca gcctccattt cctattgaag cagtttccct tctagttcat 3001 ggcatttttt atttctcttt agacatgaag agtatttctg gagaagaatc atcccatgga 3061 gtgattatga caaggcttac cgaaagtgga cacccttctt cagatgcctg gaaaggtgag 3121 aattggttat ataggaacca gaaaaaatgg gacataaatt tgccacaaga ggctttcatt 3181 cctgagacaa tttacactga ggaggaagat tttgaatgta gtgaaaataa gaaaagcttt 3241 gatattaatt cagttagctc aatttgtgct atacaagtgg gaattccttc aagaaagggg 3301 tctccaaaat gtgataagtt taaaacttac ttcaaattta atttagactc agtaggtaag 3361 caacattcag aatatgaata tgggaatgac ttgagtttga gtacagatat tcgacaccaa 3421 aaaagtcata ctacaatgaa ttcctatgaa tgttatcaat gtgggaaagc cttctgccga 3481 agttcatccc ttattcgaca tcagatcatt cacacaggag agaaacccta taaatgcagt 3541 gaatgtggga gattcttcaa ccgacgtaca aaccttacta agcatcaaaa acttcatgct 3601 gaagcaaagg cctgcacaag caataaatgt ggaaaggcct tcagtaaaag tgaagacagt 3661 aataatccaa cactccattt tggaaacaat ttctatcaat gtgttaactg tggaaaatcc 3721 ttcaaccgga gctcctctct tattcgacac caaatgattc acacgggaga gaaaccattc 3781 aaatgtaagg aatgtagtaa agccttcaac aggagttcaa accttgttaa acatcaaaaa 3841 ctgcatactc gagataagtc cgggattcca actactgcaa gcgagaatct ttattttcag 3901 ggcgccgcca aattcaaaga aaccgctgct gctaaattcg aacgccagca catggacagc 3961 ggaggtggag gttcgagcgg tccctcgggt tcgtcgagcc tggaagttct gttccagggg 4021 cccctgtcgt cgagcggtcc ctcgggttcg atggtgagca aaggcgagga gctgttcacc 4081 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg 4141 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc 4201 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag 4261 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc 4321 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc 4381 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac 4441 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac 4501 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac 4561 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc 4621 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa 4681 gaccccaacg agaagcgcga tcacatggtc cttaaggagt tcgtgaccgc cgccgggatc 4741 actctcggca tggacgagct gtacaagtaa tgatctagag tcgagttaat taagaattcc 4801 gccccccccc ctctccctcc ccccccctaa cgttactggc cgaagccgct tggaataagg 4861 ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag 4921 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc 4981 caaaggaatg cagggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg 5041 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag 5101 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca 5161 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt 5221 caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc 5281 tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa 5341 ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca acctgttaca 5401 ttgcacaaga taaaaatata tcatcacgaa cagtaaaact gtctgcttac ataaacagta 5461 atacaagggg tgttatggga tcggccattg aacaagatgg attgcacgca ggttctccgg 5521 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg 5581 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc 5641 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga 5701 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc 5761 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag 5821 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat 5881 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg 5941 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca 6001 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct 6061 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg 6121 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg 6181 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc 6241 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagt caaaaagaaa aaatgaaaga 6301 ttaccttcag tcagaatgca gaactcattt aacatcatga atttatgctg gataaaatct 6361 catgaatata atgtaagaaa acatttgtca gatttttctt taaatgatat cacagaatta 6421 atgttggata gaaatatcat tggatagaaa tctgtttgaa ggaattttcc tggttttcag 6481 atgaagccat aaggctttgg agttaaacca cagtatcacc atacaagtat ttgtttatca 6541 ttattttgat atccattacc ctcacctctc cctagttcat aaataggtct atagcatact 6601 aatatccacc tgatttattg acgaagtaga cattaggcaa agccaaacaa tactcttgga 6661 gttgatattt aataaaactc agccctttta agaaggtcac aagaaatcat tagctcatgc 6721 gaatataaga gaatacttct aaggacagaa atctctgaat caatggctaa gacaacagta 6781 acagccattt actccactcc tgacaatacc ctttgttgtc ttgctgttga atggagacta 6841 gagttgggaa agggttatca actgcaatgg gagaagtatg tattgctgtt aatctatatg 6901 tattcttaaa atctgcattt gtttatagca ggtccttgaa aaatgttttt gttttgttat 6961 aatgttataa ttgttatgat gttgatgaga aaaatatcga ttcccagcca gggacactgt 7021 gtggagttcg cacactctcc ctatgtctcc gggtgctcca gtttcaaacc ccaaagatgt 7081 atacattagg ttaattgaca tatctaaatt atctcagtct gagtttgtga ggaagtgtgc 7141 cctgcagtgg aatggctcct aatcagggtt tgttcccacc attgccctga gctgctagga 7201 tgcgctctgt ccacccatgg ccctgtactg gaataacagg gtaaataatt atcttatctg 7261 tttttattaa tccgtcttaa atgtgtatat agctcagatt tattttaatg tctgatattg 7321 gaagcgtttg gggtctttag aagtttgatg atgttgtgac cagacatttg cc //