LOCUS Exported 7039 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 7039) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 7039) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7039 /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 3676..3696 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3709..3753 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3793..3816 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 3844..4563 /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(3889..3910) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4150..4169) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4601..5186 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4767..4784) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4994..5013 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5268..6071 /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(5331..5350) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5941..5960 /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 gcagggarag catttagccg tggctcacac 2701 cttattcaac atcagaaaac tcacactggt gaaaaaccct ttggatgtaa ggaatgtggg 2761 aaggccttca gccgtgcctc acaccttgtt cagcatcaga gaattcacac gggtgagaaa 2821 ccttatgact gtaaggactg tgggaaggcc tttggtcgta catcagaact tattctacat 2881 cagagacttc atactggtgt caaaccttat gaatgtaaag aatgtgggaa gacctttagg 2941 cagcattcac agctgattct gcatcaaaga actcacacag gcgagaaacc ctacgtatgt 3001 aaagactgtg gcaaggcttt cattcgtggc tcccaactca ctgtgcatcg gagaatccac 3061 acaggggcca gaccctatga gtgtaaagaa tgcgggaaag cctttagaca gcactcacag 3121 ctgactgtac accagcgaat ccacactggt gagaaaccct acgagtgtaa ggaatgcgga 3181 aagggcttta ttcacagctc agaagttact cgacatcaaa gaattcattc tggggagaaa 3241 ccctatgagt gtaaggaatg tgggaaggcc ttcagacagc acgcacagct cacacgacat 3301 cagagagtcc atactggcga cagaccctat gaatgtaagg actgcgggaa ggcatttagt 3361 cgtagctcat acctcattca acatcaaaga attcatacag gtgacaaacc ctacgaatgt 3421 aaggaatgtg ggaaggcctt tattcgtgtt tcacaactga ctcatcatca gcgaattcac 3481 acttgtgaga aaccctatga atgcagggag tgtggaatgg cctttattcg tagttcacaa 3541 cttaccgaac atcagagaat tcatcctggt atcaaacctt acgaatgtag agagtgtggg 3601 caggcattta ttcttggctc acaactcatt gaacactaca gaattcatac tggtgggatt 3661 ccaactactg caagcgagaa tctttatttt cagggcgccg ccaaattcaa agaaaccgct 3721 gctgctaaat tcgaacgcca gcacatggac agcggaggtg gaggttcgag cggtccctcg 3781 ggttcgtcga gcctggaagt tctgttccag gggcccctgt cgtcgagcgg tccctcgggt 3841 tcgatggtga gcaaaggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg 3901 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc 3961 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc 4021 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg 4081 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc 4141 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc 4201 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg 4261 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag 4321 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc 4381 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac 4441 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg 4501 gtccttaagg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag 4561 taatgatcta gagtcgagtt aattaagaat tccgcccccc cccctctccc tccccccccc 4621 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt 4681 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt 4741 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcagggtc tgttgaatgt 4801 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct 4861 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt 4921 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt 4981 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa 5041 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta 5101 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa 5161 aacacgatga taatatggcc acaacctgtt acattgcaca agataaaaat atatcatcac 5221 gaacagtaaa actgtctgct tacataaaca gtaatacaag gggtgttatg ggatcggcca 5281 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct 5341 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc 5401 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg 5461 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg 5521 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc 5581 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc 5641 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg 5701 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc 5761 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg 5821 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc 5881 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag 5941 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg 6001 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg 6061 agttcttctg agtctagaaa gccttgagtg taagggataa agataagcct ttatgtggag 6121 ttcacatctg gctcagtact aggtaatttt taatgtagtg taattaatgt cagaaaacac 6181 ctgtcacttc catcatagaa catcagaaaa ctcacaccca aagaaaatta aataaatgtg 6241 ggaattcttt ttgcctcact cactgtttcc taagcatcag ataatctatg ctaaaaaaaa 6301 aatgaatata gaaaagctgc ttcttgccat tcaaaatatg ttaaatttct gacaattcct 6361 aatacaaaaa tgacccgatt aaaagattct gtggccaggc gtggtggctc atgcctgtaa 6421 tcccagcact ttgggaggcc aaggccggca gatcatttga ggtcaggagt ttgagaccag 6481 cctggccaat gtggcgaaac cccatcacta ctaaaaatac aaaaattagc caggcatggt 6541 ggcgggtgcc tgtaatccca gctactcagg aggctgaggc aggaaaaccg tttgacccca 6601 ggaggcagag gttgcagcaa gccgagatcc caccattgca ttccagcctc gttgacaagg 6661 acaaaactct gtctcaaaaa aaaaaaaaaa aaaaaaggtt ctggctgggc atagtggctc 6721 acgcctgtaa tcccagcact ctgggaggcc gaggcgggcg gatcacaagg tcaggagatc 6781 gagaccatcc tggctaacac agtgaaacct gtctcaacta aaaactatga aaaaattagc 6841 tgggcctagt ggcgggcacc tgtagtctca gctactcggg aggctgaggc aggagaatgg 6901 cgtgaaccca ggaggcggag cttgcagtga gccaagatcg caccactgca ctccagcctc 6961 ggcgacagag caagactcca tctcagaaaa aaaaaaaaaa aaaagattct atgactgtgc 7021 ctttcatcat ggcgtatac //