LOCUS Exported 5069 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor THRB with GFP. ACCESSION . VERSION . KEYWORDS pTHRB-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5069) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 5069) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5069 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 31..619 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 520..539 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 1674..1694 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 1707..1751 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 1791..1814 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 1842..2561 /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(1887..1908) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2148..2167) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 2599..3184 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(2765..2782) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 2992..3011 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 3266..4069 /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(3329..3348) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3939..3958 /label=Neo-F /note="Neomycin resistance gene, forward primer" ORIGIN 1 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg 61 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct 121 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct 181 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct 241 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg 301 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc 361 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga 421 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 481 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta 541 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 601 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggggaa gcccagtagt 661 ctgcacagga cttgggaaga tttgctgggc tttccagggt cccggtgtct tctcactcca 721 gctcccacta tagctgttct cttctctgca tggggtgact ctggcacagc tcccctcccc 781 ttttcatttc aattccaaaa tcatgactaa tacaaggctg ctactggtag caattttact 841 aaaggcataa gggaagggaa aggccagttc attgcagcct ggaagacaac acaaaattaa 901 acatggtgtt tcttggtcct caaggaaatg tcttactatt ccgcgttcct cctttggcct 961 caggcctgct gcaatgtccc cagtcccttc atatatctcc cccttctatg catgtccacg 1021 ctcatcctct cctctctgcc aggcagccag tgaaaattca aaattgttcc cctcatcttg 1081 cagaatattt cagctatgtc aggcttctcc ttctgtaccc taggagggag gcatcacctt 1141 tcacactcaa gaaggtaatt cacactcaag ctaaactgtc taaacagaga tatttatctg 1201 aacaaagtac tgcctctctt catataaatg atgccactcc cctggtacct ggtccctgga 1261 gcaccagagt tcacgctgca tccatgctca tggctaagag ggaagaccct agatctgccc 1321 gccccccaat aaaggcctgg aattggacaa agcaagcctt cccccttcca tctctgaatc 1381 aatgtccatc ttctctcttc cccgcagatc gcccggggct tgcctgtgtt gagagaatag 1441 aaaagtacca agatagtttc ctgctggcct ttgaacacta tatcaattac cgaaaacacc 1501 acgtgacaca cttttggcca aaactcctga tgaaggtgac agatctgcgg atgataggag 1561 cctgccatgc cagccgcttc ctgcacatga aggtggaatg ccccacagaa ctcttccccc 1621 ctttgttctt ggaagtgttc gaggatgatt atgatattcc aactactgca agcgagaatc 1681 tttattttca gggcgccgcc aaattcaaag aaaccgctgc tgctaaattc gaacgccagc 1741 acatggacag cggaggtgga ggttcgagcg gtccctcggg ttcgtcgagc ctggaagttc 1801 tgttccaggg gcccctgtcg tcgagcggtc cctcgggttc gatggtgagc aaaggcgagg 1861 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca 1921 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt 1981 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct 2041 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt 2101 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact 2161 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga 2221 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca 2281 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca 2341 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca 2401 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg 2461 ccctgagcaa agaccccaac gagaagcgcg atcacatggt ccttaaggag ttcgtgaccg 2521 ccgccgggat cactctcggc atggacgagc tgtacaagta atgatctaga gtcgagttaa 2581 ttaagaattc cgcccccccc cctctccctc ccccccccta acgttactgg ccgaagccgc 2641 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt 2701 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt 2761 tcccctctcg ccaaaggaat gcagggtctg ttgaatgtcg tgaaggaagc agttcctctg 2821 gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca 2881 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg 2941 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc 3001 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct 3061 gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta 3121 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac 3181 aacctgttac attgcacaag ataaaaatat atcatcacga acagtaaaac tgtctgctta 3241 cataaacagt aatacaaggg gtgttatggg atcggccatt gaacaagatg gattgcacgc 3301 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat 3361 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt 3421 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg 3481 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag 3541 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc 3601 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc 3661 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga 3721 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga 3781 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg 3841 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg 3901 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc 3961 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc 4021 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgat agactgactg 4081 gattcattct cataattcct acagcactac tgggtgtcat ttcattccat tgcctagctc 4141 ttttttgttt gtttctttgt gttgggaggg attatttggg agggaaaagg gaagtagtcc 4201 ttggcataga catggatgaa attgcccctt gaatgcgggt acttgaaact attgcatttc 4261 gttctccggt cctgtgatgt gaatgctctg aaggttttat ggttgtggag gtggggtggg 4321 ggacaatcat taactcacca gcaccaagca tcaccagctc ccacccgtcc ctggtccaag 4381 acttgagtca gcaaaatggc gccacaggac actaaagaag ccttaaaacc aagataatac 4441 gaccacctcc acccaatcct gatgttcgca gggctgaagt taacagagca cagaccacct 4501 ttagttagat gtggctttca gccttttagg gaaagactcg aacaaatttt catctattca 4561 agagcatgct gtttgcagca ctctttatgg tctcctagga gcataacttg gaatgtacag 4621 gaatatttgt gtcctaaaaa aatcagctgc ccttccaata tcatttttat aacgttatgg 4681 tttaattcaa gttatacaaa ggtgtgactc aaggatgtgc aggtattgac tgatggaggc 4741 caacaaggaa aatatttttt ttcagtgagc ccagagaaat tagaaatctt caccaatgtc 4801 agtgactgca tggtttccaa aaattcacat agtattacct cacagtagga aggcttcttt 4861 cccatttagg ctcagagaac acttttacag aaatcaaacc tatttacagg gtggcacccc 4921 ttttctgtgg catctgatct gtcatctccc aagctcctga agttctttat ccaggttgta 4981 tctttgctga gctgacaaca gatttctcta tggatcaaat attctaatca aggaaagata 5041 cggtgaaact tcagtcttct tctggagaa //