LOCUS Exported 7137 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor TFE3 with GFP. ACCESSION . VERSION . KEYWORDS pTFE3-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7137) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 7137) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7137 /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 3882..3902 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3915..3959 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3999..4022 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 4050..4769 /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(4095..4116) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4356..4375) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4807..5392 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4973..4990) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5200..5219 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5474..6277 /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(5537..5556) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6147..6166 /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 ggtggaggca gagagagagc aggccagggt 2701 ttcccttcct ggcatacttg ggggatccct gggcttccca gatcacacga tgatgatagt 2761 gataatgata aataataatc ataatattaa caaggataat aatattgcta atgtacatag 2821 catttatctt gtgctctgta ctgtgacaag agctttatat ccactaactt attcaatcct 2881 cagaattcta tgagataggg gccattattt tatttattta tttatttatt tatttttgag 2941 atggagtttc gctcgttgcc caggctggag tgcaatggcg caatctcggc tcaccgcaac 3001 ctccgcctcc cgggttcaag caattctcct gtctcagcct cctgagtagc taggattata 3061 ggcatgcacc accacaccca gctaattttt gtatttttag tagagatggg gtttctccat 3121 gttggtcaag ctggtctcga actcctgacc tcaggtgatc cacctgcctc ggcctcccaa 3181 agtgattaca ggcgtgagcc accgcgccca gcctaggggc cattattata tacccacttt 3241 acagtttgcg actctgaagc ttcagagatg ttaagcacct tgcctaaagt tttggaacta 3301 gtgagtagca gaagctgaat tccaacccag ggaacctacc cttttcaacc tttatactcc 3361 tctgcctctg agggactctg aaccctactt atcctgcacc caacccctat tttttacagg 3421 aactagaact gcaggcccag atccatggcc tgccagtacc tcccactcca gggctgcttt 3481 ccttggccac gacttcggct tctgacagcc tcaagccaga gcagctggac attgaggagg 3541 agggcaggcc aggcgcagca acgttccatg tagggggggg acctgcccag aatgctcccc 3601 atcagcagcc ccctgcaccg ccctcagatg cccttctgga cctgcacttt cccagcgacc 3661 acctggggga cctgggagac cccttccacc tggggctgga ggacattctg atggaggagg 3721 aggagggggt ggtgggagga ctgtcggggg gtgccctgtc cccactgcgg gctgcctccg 3781 atcccctgct ctcttcagtg tcccctgctg tctccaaggc cagcagccgc cgcagcagct 3841 tcagcatgga agaggagtcc gggattccaa ctactgcaag cgagaatctt tattttcagg 3901 gcgccgccaa attcaaagaa accgctgctg ctaaattcga acgccagcac atggacagcg 3961 gaggtggagg ttcgagcggt ccctcgggtt cgtcgagcct ggaagttctg ttccaggggc 4021 ccctgtcgtc gagcggtccc tcgggttcga tggtgagcaa aggcgaggag ctgttcaccg 4081 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt 4141 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca 4201 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt 4261 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg 4321 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg 4381 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact 4441 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg 4501 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca 4561 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg 4621 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag 4681 accccaacga gaagcgcgat cacatggtcc ttaaggagtt cgtgaccgcc gccgggatca 4741 ctctcggcat ggacgagctg tacaagtaat gatctagagt cgagttaatt aagaattccg 4801 cccccccccc tctccctccc cccccctaac gttactggcc gaagccgctt ggaataaggc 4861 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg 4921 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc 4981 aaaggaatgc agggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga 5041 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg 5101 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag 5161 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc 5221 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct 5281 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac 5341 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa cctgttacat 5401 tgcacaagat aaaaatatat catcacgaac agtaaaactg tctgcttaca taaacagtaa 5461 tacaaggggt gttatgggat cggccattga acaagatgga ttgcacgcag gttctccggc 5521 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga 5581 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct 5641 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac 5701 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct 5761 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt 5821 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt 5881 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt 5941 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag 6001 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt 6061 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg 6121 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg 6181 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg 6241 catcgccttc tatcgccttc ttgacgagtt cttctgagtc tcaggcctca cccctcccct 6301 gggactttcc cacccaggaa aggaggacca gtcaggatga ggccccgcct tttcccccac 6361 cctcccatga gactgccctg cccaggtatc ctgggggaag aggagatgtg atcaggcccc 6421 acccctgtaa tcaggcaagg aggaggagtc agatgaggcc ctgcaccttc cccaaaggaa 6481 ccgcccagtg caggtatttc agaaggagaa ggctggagaa ggacatgaga tcagggcctg 6541 ccccctgggg atcacagcct cacccctgcc cctgtgggac tcatccttgc ccaggtgagg 6601 gaaggagaca ggatgaggtc tcgaccctgt cccctaggga ctgtcctagc caggtctcct 6661 gggaaaggga gatgtcagga tgttgctcca tcctttgtct tggaaccacc agtctagtcc 6721 gtcctggcac agaagaggag tcaagtaatg gaggtcccag ccctgggggt ttaagctctg 6781 ccccttcccc atgaaccctg ccctgctctg cccaggcaag gaacagaagt gaggatgaga 6841 cccagcccct tcccctggga actctcctgg ccttctagga atggaggagc caggccccac 6901 cccttcccta taggaacagc ccagcacagg tatttcaggt gtgaaagaat cagtaggacc 6961 aggccaccgc tagtgcttgt ggagatcaca gccccaccct tgtccctcag caacatccca 7021 tctaagcatt ccacactgca gggaggagtg gtacttaagc tcccctgcct taacctggga 7081 ccaacctgac ctaacctagg agggctctga gccaaccttg ctcttgggga aggggac //