LOCUS Exported 7010 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor ZBTB1 with GFP. ACCESSION . VERSION . KEYWORDS pZBTB1-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7010) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 7010) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7010 /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 3653..3673 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3686..3730 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3770..3793 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 3821..4540 /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(3866..3887) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4127..4146) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4578..5163 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4744..4761) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4971..4990 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5245..6048 /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(5308..5327) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5918..5937 /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 tcggtacccg aaaacacagg aatgtctagg gccaagggat 2701 gaccaaaatt taaaagatta tttcatatca gagtcaggga tattaataat tttatgtgag 2761 aaaatttgca gtttgttttc tcctatttga ggaaggaaca gttaattata cagcttaaaa 2821 agcccctact tacttaaaaa gttaacatgt ttttattctc cttaaactaa ctgctcatat 2881 caaacacaat caggagagct catttttaga tgatagaaac attaattcta agcatccaga 2941 ttatgaacca tctttttttt caaatgctta tgtttaaaaa atttaaaaaa aattaaggat 3001 cttattatga gggaaaagca ctgttgccaa aaatataatt ttactcagaa tcactagggc 3061 agattaggag gcttataagc ctaggaattt gcaggaaaac ccctcctctt cttttgtgcg 3121 tgtatgtgtg tgtgcgtgta tgcttcattt tcatttgcag agtggctgaa gacagtaaaa 3181 agaatatgaa ctttaaaaaa attgttagat ttatgtttca atcagagatt tgtcattaac 3241 aagttaaagt acattgtaaa atcattttat ttgaaggaaa acacttctta actgcttcct 3301 tcctccctac tttcctctat ttgtctgacc cctatctttt tttatgttat atatttatat 3361 gtcctgctga tagattaata agaacatctt atatttgaat ggtacgagtt acattcggga 3421 aataattaaa tttaaattct aggggctatg cttgtgtttc tattctctgt tctgttaaca 3481 gtgtgttggc ctagtgccaa agccaagaaa ttctgatcta agaattatgt tgtatcttgt 3541 agagcagtca acttgtaata ttatttttct tgagttttgt ctttttctgt gtggcagtgg 3601 tgaaataggg ccttctaaac ctgtggagaa ggggattcca actactgcaa gcgagaatct 3661 ttattttcag ggcgccgcca aattcaaaga aaccgctgct gctaaattcg aacgccagca 3721 catggacagc ggaggtggag gttcgagcgg tccctcgggt tcgtcgagcc tggaagttct 3781 gttccagggg cccctgtcgt cgagcggtcc ctcgggttcg atggtgagca aaggcgagga 3841 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa 3901 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt 3961 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta 4021 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc 4081 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta 4141 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa 4201 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa 4261 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa 4321 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac 4381 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc 4441 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc cttaaggagt tcgtgaccgc 4501 cgccgggatc actctcggca tggacgagct gtacaagtaa tgatctagag tcgagttaat 4561 taagaattcc gccccccccc ctctccctcc ccccccctaa cgttactggc cgaagccgct 4621 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg 4681 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt 4741 cccctctcgc caaaggaatg cagggtctgt tgaatgtcgt gaaggaagca gttcctctgg 4801 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac 4861 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg 4921 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct 4981 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg 5041 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag 5101 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca 5161 acctgttaca ttgcacaaga taaaaatata tcatcacgaa cagtaaaact gtctgcttac 5221 ataaacagta atacaagggg tgttatggga tcggccattg aacaagatgg attgcacgca 5281 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc 5341 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 5401 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg 5461 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg 5521 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct 5581 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct 5641 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 5701 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa 5761 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc 5821 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt 5881 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct 5941 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 6001 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagt ctgagagatc 6061 tgaatttgga gaggacatgg ggaagaatca atcttcaaca gataatcttt aagtacagac 6121 ccaggaatat cagatctgaa gtggcaccat cttttctgtc ttcctgacaa acttccatca 6181 acccaaaaag ttccagttgc aaagatccaa gaaaactcag taaagatcac tttggaatca 6241 ctttgttgta acaaaattga ttcatggtga ctctaatatt tgtgtaaatc tgctgaagaa 6301 aagtaactag aaaattcata caacatcagt ggttacagct atactaaaat aaaactttat 6361 atgctaatta tcttcaaggg tctttctaaa ataaaagggg ttgttaggct actgcaaata 6421 taaagagaca taatcacaaa tctatttata tagatataaa taattctaaa cttaaaatca 6481 ctagaattat agcttaaata ttatgaaatg tacttagtca acttaaaaac ttacaccaaa 6541 aatctgtagt ttagagtgta agaagtaatg ggtacagaga aaaaaaattt aagaccaata 6601 ttttaacagg ataatttggt tattactcac tatagtaaaa cacttgccct tcttcctttt 6661 ccactgtata tgccagtcac aatcttgatc atttatatga atgaaactcc cttttaaacc 6721 tgttaacttg aaggagaaca gaaatggtat tacttcaata gtggtagatt tccacaaatc 6781 agacacatta ttttcttctg aacttggcaa tataacctaa taaggcagaa taagtttctg 6841 ttaaaatact tattctcaat atttatgtat aaaccatgct tctcagcttt caaatttcaa 6901 gataaaattt aagtattcag taaaatgatg gttaacatgt tttaaaaaat ctttaatgga 6961 gaagttaaat atttggtgag attttagtct ggctataaat gctttgtttg //