LOCUS Exported 7038 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor MXD1 with GFP. ACCESSION . VERSION . KEYWORDS pMXD1-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7038) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 7038) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7038 /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 3695..3715 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3728..3772 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3812..3835 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 3863..4582 /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(3908..3929) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4169..4188) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4620..5205 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4786..4803) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5013..5032 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5287..6090 /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(5350..5369) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5960..5979 /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 tcggtaccca gcttcagcga gagcagcgac acctgaagag 2701 gcagctggag aagctgggca ttgagaggat ccggatggac agcatcggct ccaccgtctc 2761 ctcggagcgc tccgactccg acaggggtga gcctctctca ctctcctccc tgtctccctt 2821 gtgctcccca accccagagc aggggttcag tactgcggac aggaggcaga gtgtaagaag 2881 aactaagaca ctgggtatga tgtagagaag tcgaacagtg tgacctccag tgacaggttc 2941 cttctccctg ctaacactta ggtgacaacc ccagaagagt ttgattggtg tcagttttca 3001 gcctagtttg ttttgttttg ttttgagacg gaatcttgct ctgtcaccca ggctggagtg 3061 cagtggtgtg atcttggctc actgcaacct ccgcctcccg ggttcaagcg attcccctgc 3121 ctaagcctcc caagcagctg ggattacagg cacacaccac aatgcctggc aaattttcct 3181 atttttgtag agatgaggtt tcaccatatt ggccaggctg gtctcgaact cctgacctca 3241 agtaattcac ccaccttggc ctcccaaagt gctgggatta caggcgtgag ccaccacacc 3301 cagccaaatt tttgtattag tagaaacagg gtttcactgt gttggcccag ctggtctcaa 3361 actcctaacc tcaagtgatc cacccgcctt tgcctcccaa agtgctggga ttacaagcat 3421 gagccaccac gcctgagctt tctgcagagc gctttcatca atgtccttct ctcttgtcct 3481 ccctgcagaa gaaatcgacg ttgacgtgga gagcacggac tatctcacag gtgatctgga 3541 ctggagcagc agcagtgtga gcgactctga cgagcggggc agcatgcaga gcctcggcag 3601 tgatgagggc tattccagca ccagcatcaa gagaataaag ctgcaggaca gtcacaaggc 3661 gtgtcttggt ctcgggattc caactactgc aagcgagaat ctttattttc agggcgccgc 3721 caaattcaaa gaaaccgctg ctgctaaatt cgaacgccag cacatggaca gcggaggtgg 3781 aggttcgagc ggtccctcgg gttcgtcgag cctggaagtt ctgttccagg ggcccctgtc 3841 gtcgagcggt ccctcgggtt cgatggtgag caaaggcgag gagctgttca ccggggtggt 3901 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga 3961 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa 4021 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag 4081 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta 4141 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt 4201 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 4261 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat 4321 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga 4381 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 4441 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa 4501 cgagaagcgc gatcacatgg tccttaagga gttcgtgacc gccgccggga tcactctcgg 4561 catggacgag ctgtacaagt aatgatctag agtcgagtta attaagaatt ccgccccccc 4621 ccctctccct ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg 4681 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga 4741 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa 4801 tgcagggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa 4861 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct 4921 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg 4981 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg 5041 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca 5101 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg 5161 acgtggtttt cctttgaaaa acacgatgat aatatggcca caacctgtta cattgcacaa 5221 gataaaaata tatcatcacg aacagtaaaa ctgtctgctt acataaacag taatacaagg 5281 ggtgttatgg gatcggccat tgaacaagat ggattgcacg caggttctcc ggccgcttgg 5341 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc 5401 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt 5461 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt 5521 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc 5581 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc 5641 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac 5701 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag 5761 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag 5821 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat 5881 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg 5941 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa 6001 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc 6061 ttctatcgcc ttcttgacga gttcttctga gtctaagaga gtgggcactg cggctgtctg 6121 cttgaaggtt ctccctgttg gttctgatta ggtaacgtat tggacctgcc cacaactccc 6181 ttgcacgtaa acttcagtgt cccaccttga ccaaaatcag ctttgtaact gttttcaagg 6241 aggtgcttag gattgtgggt ttctgattgc atccactagc ttctcttttt ctcgccataa 6301 aaatttgtct ctgagagact atacattcca atcaatttga agcatccaag aattcttaga 6361 ccgaataagc aatgtccaca tctcagcaat ccccctcttt gctctcctcg tgtcctctcc 6421 cagacctttc ttccagtttt ctggcttact gtgttacttg cctaggagga atgtccaagt 6481 gtgtcttgta cttaggaaac tgaaggaaac aaacttttga aattgagatc ctgatctcag 6541 aactccaaag taagctttga aagcagcatt taagagcact taaccatgga cctcaccacc 6601 agtgaggaag tcaggaatac ctctagaaaa cacgcccttc acaccgcgca tgctcattcc 6661 cccgacgcgc cgcgtgtgga tgggagcagt atttctcact ttaaaatgga caccttgata 6721 gtgtgtctct ggggttgcac agctcatcaa agttccaaat tgtttgttct cacaaacaaa 6781 acggtacaat ctatttttgt gcgatgttct tgggacctcg ctgtgttctg ctatctcgag 6841 gcacattctc ccctccaact ttgttaatgc tacttgtctc tggaacattt ttttttccta 6901 cctcagagat aaatgagatc tccatttccc acttaacaca gtgattatcc ttttttcccc 6961 ccccaaaata gtgacatttg gtccaattat aatctatttt cctattgaac tttcagcaat 7021 aactgagctg tggcacta //