LOCUS Exported 7461 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor CSDC2 with GFP. ACCESSION . VERSION . KEYWORDS pCSDC2-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7461) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 7461) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7461 /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..5395 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4976..4993) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5203..5222 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5477..6280 /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(5540..5559) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6150..6169 /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 gttctcacgc tcacagggcc atggcttcat 2701 cacccccgag aacgggtccg aggacatctt cgtacatgtg tctgagtgag tcccctccac 2761 ctccctgttc ttggcccctg ccctagtgtc cccaatggtg cctcaaaaat ggctctccag 2821 gcctctcctc tcactggctg aggtctgtat tcatggtgcc ttttctgtcc tctgagtagg 2881 ggctgagggt gcgttgggag aggagggctg gggccaggga gggagtgggc acgcagaccc 2941 cagaagcctg ggagccagcg gctgggccct ggatcctgtt acagcatcgt ggaggtctga 3001 gagggctagg ccagctcctg agcctcttgg cctggaaagg gccttcatgt gggtgggcca 3061 ggaggtagag atggatatca tgtctaggcc tggccccacc cacttctaat gccaggagga 3121 ccgggggagg tcactgagct tctcagagct ttagggacct ccccgtgccc tttcacatct 3181 cggtccccaa gggacccaga aagggaggta cagtttatga gtcccatttt attttatttt 3241 atttattttt tgagagggag tctcgctctg tcgcccaggc tggagtgcaa tggcacaatc 3301 tcggctcagt gcaacctcca cttcctgggt tcaagcgatt ctcccacctc agcctcccaa 3361 gtagctggaa ttacaggtgc cagccaccat gcctggctaa tttttgtact tttagtagag 3421 ccagggtttc gccatgttgg ccaggttggt gtcaaactcc tgactcaggt gatccgcccg 3481 cctcagactc ccaaagtgct gggattacag gcgtgagcca ctgtgcccag ccatgagtcc 3541 cgttttacag attcgagagc tgaggttctg agaagcaagg cctggccagg cctcctggcc 3601 tagggaggct tagggccagg ctgcccccaa ggccaggcca caggattgtc tggggcacag 3661 ggcctgctgc tgggctaata cccctcttcc tgcccgccag catcgagggg gagtacgtgc 3721 cagtggaggg cgacgaggtg acctacaaga tgtgccctat ccctcccaag aaccagaagt 3781 tccaggccgt ggaggtggtg ctcactcagc tggcccccca cactccccac gagacgtggt 3841 ctggccaggt cgtgggctcc 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 tctcccccct ccccccccct aacgttactg gccgaagccg cttggaataa 4861 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg 4921 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc 4981 gccaaaggaa tgcagggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct 5041 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac 5101 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc 5161 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta 5221 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg 5281 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg 5341 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca caacctgtta 5401 cattgcacaa gataaaaata tatcatcacg aacagtaaaa ctgtctgctt acataaacag 5461 taatacaagg ggtgttatgg gatcggccat tgaacaagat ggattgcacg caggttctcc 5521 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc 5581 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga 5641 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac 5701 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct 5761 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa 5821 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc 5881 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct 5941 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc 6001 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg 6061 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct 6121 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct 6181 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca 6241 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gtcgctgagt ggttcacagg 6301 ccagctggcc gggggttggg gagccacaca gggtgaacgg gcagcagccg gctccatgcc 6361 ccactgccct ggctgatgag tccttcggtg gcctcagtgt gcacgtctgt ctgtccgtct 6421 gtgcttgtgg ctatgagcgt gtgcctccac ccaccccacg acccgtgact actgtgcaag 6481 ctggccagga ggtaggtgga gggcaaggcc accagacctg gcttcgcgct gtccactgcc 6541 tctctcctcc cctccctgcc gcagacacgc aggacccgct cgccctcctg cttacccgtc 6601 cccacggtga ctgagctgcg agagcctgcg ctgggctcac tccctggcct ccgcccctga 6661 gctgtcccgt gtcagtccct ggcccggagc caggctctct gtgctactta catctccctc 6721 ccttggctga ggtcccccgc cacagcctgg acccttccct cagaagcctg gccgatgggt 6781 gcttttgggg agcctctcag cctctggcag gggaggagga ctgacacgga ggcacacagg 6841 cctctcactg ccgaggccag ccttccttcc tgctctgctc tctcagtgcc atggagaggc 6901 aggaagaggg gcctgagggc ggaaggggtt tggggctccc atttcgccgg ccagtccctc 6961 ctcctcagcc tggcagtggc gctgggctcc ttcccctggg ctgtactgag ccgagcccag 7021 gggtttgcag agggtggggg tccatctctc cagcttggct gcagacctcc tttaccctga 7081 ctcacaagcc ccactgatgc tctgggccat gcccggctga cggctgctgt ggatgccgcg 7141 agggacggac acacgtccgg ggcacccacg aggagggccc ccagcctggg agactggccc 7201 tgcggcctcc acgtcaaact ctcttcccaa agccctaaca gaccagtggc cgaggtgtgg 7261 ctcttattgc atccatccct gaagatgtgt ggctgttgct gtcacctgga gtccttttgg 7321 ggccaagatg tgtgtgcacc cggggtcgtg gccattcact cccaggcagg ggtgagggtg 7381 gcctggccca ggaggccagg aaggagggcc ctgtctgcct ccacctctgg gtgcaccccc 7441 tgcctaccac cctcccttct a //