LOCUS Exported 7387 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor NR2F1 with GFP. ACCESSION . VERSION . KEYWORDS pNR2F1-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7387) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 7387) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7387 /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 3876..3896 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3909..3953 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3993..4016 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 4044..4763 /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(4089..4110) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4350..4369) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4801..5386 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4967..4984) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5194..5213 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5468..6271 /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(5531..5550) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6141..6160 /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 actccccacc acccacatcc ccccctaccc 2701 ccccagttct gaagcagcag cagctgctgc ctgtagccta gctttgtgtc tgattgctga 2761 agatgtcgcc tttctgcagc ctggagtaac tgagtcatat tcttcctgat tattatacag 2821 cttacagaga tgtgaccaat tttcaattac ttgatctaag gtatagatag aagtgtacac 2881 cctcaccaag gatttgctgc agagtccttt cttgcataga taacggatct gaaatcctat 2941 attgctaaag cgtgggcttc atctgattcc aggcactgcc agcccatgat tccttacccc 3001 ctcccatgtg tttgttcact acacaatcta ggttctcctg gaggtttctg gttggggtgg 3061 tttgggtttg agaggggact ccaggaatcc tttaagaact gccttcaggc tgcatatttc 3121 ctaaggggta ctgaacaccc ccagatcaga ggcaaatggg gcaaaagtta atgagcacgg 3181 ccagaaagat gctcccttgc aggccgagga caggttgtta agcgcagcaa acatggttcc 3241 agcattgtac attatttaat ccaaaggtga aagaggaaaa acaaagggac tagagaggtt 3301 tctgcctctg catgtgtgtg cctctctctc cagctcccct aggcttggtg ggggtttgat 3361 ttatttttat ttgtattgta ttgggggcgg gggaggaggg aatgaagaag gggatggaga 3421 ggtataggag ggtgaatttt tcttttctct tttacttctt ttttattttc cattttctcc 3481 ttaaaaaaaa ttgatggctt attttgcctt tgctatttgt cagcctaacc gtgtgctccc 3541 tttccctgtc tctccctcct gtggctgctt gggcagacgc ctgtggcctg tcggatgcgg 3601 cccacatcga gagcctgcag gagaagtcgc agtgcgcact ggaggagtac gtgaggagcc 3661 agtaccccaa ccagcccagc cgttttggca aactgctgct gcgactgccc tcgctgcgca 3721 ccgtgtcctc ctccgtcatc gagcagctct tcttcgtccg tttggtaggt aaaaccccca 3781 tcgaaactct catccgcgat atgttactgt ctgggagcag cttcaactgg ccttacatgt 3841 ccatccagtg ctccgggatt ccaactactg caagcgagaa tctttatttt cagggcgccg 3901 ccaaattcaa agaaaccgct gctgctaaat tcgaacgcca gcacatggac agcggaggtg 3961 gaggttcgag cggtccctcg ggttcgtcga gcctggaagt tctgttccag gggcccctgt 4021 cgtcgagcgg tccctcgggt tcgatggtga gcaaaggcga ggagctgttc accggggtgg 4081 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg 4141 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca 4201 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca 4261 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct 4321 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg 4381 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg 4441 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata 4501 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg 4561 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc 4621 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca 4681 acgagaagcg cgatcacatg gtccttaagg agttcgtgac cgccgccggg atcactctcg 4741 gcatggacga gctgtacaag taatgatcta gagtcgagtt aattaagaat tccgcccccc 4801 cccctctccc tccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt 4861 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg 4921 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga 4981 atgcagggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa 5041 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc 5101 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac 5161 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag 5221 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc 5281 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg 5341 gacgtggttt tcctttgaaa aacacgatga taatatggcc acaacctgtt acattgcaca 5401 agataaaaat atatcatcac gaacagtaaa actgtctgct tacataaaca gtaatacaag 5461 gggtgttatg ggatcggcca ttgaacaaga tggattgcac gcaggttctc cggccgcttg 5521 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc 5581 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg 5641 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt 5701 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg 5761 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat 5821 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca 5881 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca 5941 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa 6001 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa 6061 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc 6121 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga 6181 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc 6241 cttctatcgc cttcttgacg agttcttctg agtcaccttg ggcgcttccc acctgccccg 6301 tccccctaga gactcagagg acccacctgg gccaaggact ccaaagccgc ggggacaccg 6361 ggaagtgcag cgggccaggc aggctgggtg ggagggagga gggccgagac aggagcagcc 6421 cacccagcag aaatacaatc cgagctacaa agcatgggaa aaagagactc ttttaggatc 6481 agatctgtga gcacgttggc gaggaaaaac aaaacaaaca aaaaaaagaa ccttgtgtct 6541 gtctggtgaa aaaaagaaaa acaaattgga agagaggacc atgagaattt taataaaaca 6601 gaaggaaact aatggacctt ccaggattta ttgtggacgg atgtggatat attctgtaca 6661 ggaacaacac atatggaagt ggactgaagc ctatgtagaa acacacacac actgaacatt 6721 gttattcatt ttgtaaaata ctagtcttta ttttcatttt ttgtaaaatt taaacatcgt 6781 atgcgcataa agaaaaagga aacaagaatt aggggaaaat aacattttcc aaataattat 6841 aaaaaattgt cctgtgtcta tgtatctata tctgttttgt atttttttct ggttccaaac 6901 cagatttcct gtgattctat actaataatt tttgatataa ccctttgctt cttataatga 6961 gtgcgatata tgttgtcgag gctgttcttc aagaattaaa attgaagtga aaatttaaac 7021 aaaaataaaa gaatttagca aaacccatgt gtctggttgc ttgatagatg ttatccgtgg 7081 aagagaactc cttggttaat ttccacctca actattatta gaattgaagg ttttggtgtg 7141 taaaagggga gacttttaaa gtctactaca aagttggtta gattcatttc caaaaaccaa 7201 gctgctacct tggttttgta aatgcggagg gccaggctct acatgggctg gggtccgcgg 7261 ggaggcaggg cgccttggtc gctgacttca gccgttgaga cctggtgggc tgcgggcggt 7321 cccgtgctca gttgggccag ggagtggtgt aggcagggtc gcgcgcccgg taagtctcca 7381 ggcacca //