LOCUS Exported 7071 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor ZNF174 with GFP. ACCESSION . VERSION . KEYWORDS pZNF174-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7071) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 7071) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7071 /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 3694..3714 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3727..3771 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3811..3834 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 3862..4581 /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(3907..3928) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4168..4187) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4619..5204 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4785..4802) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5012..5031 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5286..6089 /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(5349..5368) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5959..5978 /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 taccaggaaa gcctttggca gaacaatgta 2701 ttaggtaaga aggactttta acagcccctc accatactgc ttcctgttcc tacagggcct 2761 atttccagca gaagtaggtg gggggtggtg tgtagagtag gcatctgatg gcccagtgtt 2821 gtgactgacc ctgatatttg tgccagcaac taccctgatg actttgactc tggcccttga 2881 gaaaggccca agctttaggg gaagccactc ggctgatttg tagcatcctc attcctgtca 2941 ttcttcacgt tctccccttt acttcctttg agggtttcag tggtttctat gaaagttgac 3001 ctctagatgg ttatcagcaa ttcccaaggt aaacgggtca ttatacaaca gtgagagatc 3061 aagcttccct cagtcctgat ggatggaatt aatggatggg gcttgttcct aggtggccgt 3121 ttgtatgcag gggcaaaagg tgctcttgga gaaaactgga tctcagcttg gagaacagga 3181 actgccagac tttcaaccgc agactcctag gagagatctc agggagagct ctccagcaga 3241 gccttcccag gcaggagctt atgaccggct gagcccccat cattgggaga aatccccact 3301 cctccaagaa ccaaccccca aattggctgg gacaggtaaa cactctgcct tttctctccc 3361 tctcatcagc cctttatctc catcaatgta tctcatggtt ctgcaaaggc atagaccaca 3421 tgtgtgtgtt tttttaaatg ttatatataa tagtagatga ttattcacta aacaaatttc 3481 actaattagg atggtggtga tactacaagt aagtatgttt ttatcgtttt ctgttaatga 3541 aaatttaaga atccagtgca tgacagggtt gtacctactt ttctgtccgt ttcagaactt 3601 cttatagaaa agacagatcc aaatatggcc acagatgaac ttccatgcaa gctatggctg 3661 agtttcattg ctgggattcc aactactgca agcgagaatc tttattttca gggcgccgcc 3721 aaattcaaag aaaccgctgc tgctaaattc gaacgccagc acatggacag cggaggtgga 3781 ggttcgagcg gtccctcggg ttcgtcgagc ctggaagttc tgttccaggg gcccctgtcg 3841 tcgagcggtc cctcgggttc gatggtgagc aaaggcgagg agctgttcac cggggtggtg 3901 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag 3961 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag 4021 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc 4081 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac 4141 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg 4201 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag 4261 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc 4321 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag 4381 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc 4441 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac 4501 gagaagcgcg atcacatggt ccttaaggag ttcgtgaccg ccgccgggat cactctcggc 4561 atggacgagc tgtacaagta atgatctaga gtcgagttaa ttaagaattc cgcccccccc 4621 cctctccctc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc 4681 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa 4741 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat 4801 gcagggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac 4861 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg 4921 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt 4981 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg 5041 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac 5101 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga 5161 cgtggttttc ctttgaaaaa cacgatgata atatggccac aacctgttac attgcacaag 5221 ataaaaatat atcatcacga acagtaaaac tgtctgctta cataaacagt aatacaaggg 5281 gtgttatggg atcggccatt gaacaagatg gattgcacgc aggttctccg gccgcttggg 5341 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg 5401 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg 5461 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc 5521 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg 5581 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca 5641 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc 5701 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg 5761 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg 5821 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata 5881 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg 5941 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat 6001 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct 6061 tctatcgcct tcttgacgag ttcttctgag tctaaaatgc tcctctcttt agaggggcct 6121 ggtagaaata aaaacccacc ctatatctta tatctttgtc ctcctctgtg atgttgcttg 6181 tactttagat cctcaaatct atttcccttt ctctggtgtg atacttgtgt aactgctggc 6241 cttgttctac acagtcctgt gatctctaag cctgaatgac agtgacattt tggccaagcc 6301 tggtgatgtg aatacaagat acagtgagca cgttagggag gagggttaca agacgacaaa 6361 aatcctcagt ttcttaatat ttgctacatc caatgaggta cttcgtgttt tagtctgttc 6421 aggctgctaa tataaaatgc tagagactag gtagtttata aacaacagga gtttatttct 6481 cacagttcta gaggctagga agtctgagtt caaggtgcca gcatggttgg gttccagtga 6541 ggcccctctt ccaggtcaca gactgcctac ttctcagtcc tcatgtggca gaaggggaag 6601 acaggctccc gtggggctct ttcatgaagg cactaatccc attccagtga gggcagcacc 6661 ctcatgacct aatcacctcc caaaggcccc atgtcctaac actattacct tgggagttag 6721 aatttcatat gaatttggag gaacataaac atttagcccc tggcaaccac agtctacttt 6781 ttgtctctag aattgcttat tctgaacatt ttatacaaat ggaatcacac aggccgggca 6841 cagtggctca cgcctgtaat cccagcactt tgggaggcca aggcaggcgg atcacttagg 6901 ctcaggagtt cgagaccagc ctgggcaaca tggcgaaatc ctgtctctac aaaaaatgca 6961 aaaattagcg gggcatggtg ctgcacacct gtaatcccag ctgcttgggg gagctgaggc 7021 aggaggatca cttcaacctg ggaggtcgag gctgcagtga actgagatcg t //