LOCUS Exported 6827 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor TBPL1 with GFP. ACCESSION . VERSION . KEYWORDS pTBPL1-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6827) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 6827) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6827 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(19..36) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(19..35) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(28..50) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(244..263) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 363..385 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(423..441) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 509..613 /gene="bla" /label=AmpR promoter CDS 614..1474 /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 complement(832..851) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1645..2233 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2134..2153 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2387..2404 /label=L4440 /note="L4440 vector, forward primer" protein_bind 2521..2542 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2557..2587 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2595..2611 /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)." primer_bind 2600..2622 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2619..2635 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2619..2635 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 3543..3563 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3576..3620 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3660..3683 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 3711..4430 /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(3756..3777) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4017..4036) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4468..5053 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4634..4651) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4861..4880 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5135..5938 /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(5198..5217) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5808..5827 /label=Neo-F /note="Neomycin resistance gene, forward primer" ORIGIN 1 ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg 61 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga 121 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct 181 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct 241 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc 301 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt 361 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa 421 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac 481 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 541 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 601 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc 661 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga 721 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga 781 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg 841 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc 901 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac 961 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact 1021 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca 1081 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg 1141 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact 1201 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg 1261 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg 1321 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat 1381 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc 1441 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat 1501 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt 1561 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc 1621 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt 1681 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac 1741 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt 1801 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 1861 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 1921 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 1981 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 2041 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt 2101 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc 2161 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg 2221 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc 2281 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc 2341 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag 2401 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca 2461 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat 2521 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg 2581 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga 2641 ttacgccaag cttaggatgt gtggttagga ctttcagaga gtttacagtg ggtggatttg 2701 ttagtttcct gggtctttca aaagaggtta agtgtgtatt ggtgaagcca tttttattca 2761 aaattttatg agaatagttt tcattgtcct tttttatgtt tctttttagc gatttaaatg 2821 aaagattaca tttttataaa gttgttcaag aattagcata atgcatggtc acctcgtagc 2881 cttgtgcttt ttttgttttc tattgacttg tatacgctct gagggtgaaa tgaccagatt 2941 ggtaataaat atgtggatac ccagtggtgg taaatctgtg gggattttta aaccagcatt 3001 ctggtttccc aacattttgg tagcagcttt gtttcactct gctgtgttcc tccatatccc 3061 tgagcctctg ttgtagccta accagaagcc agatgatagt gacttaaagg atagctgtta 3121 tttagtccca tttgttactc cttcctttca tgtacatatt atattacagt gctttttata 3181 ttagtccatg atagtttgta tgccctaaaa caaagtgaat aacctcattt tgtgtttcat 3241 atgtccttta attgcccaga acaaaatctg tattactttt acttattatg gctatgtgta 3301 taattatatg tctagttttc ttttttttaa cagttaactt gatattctat accacttgaa 3361 tatttttaat tctggaaaat cagtttttcc tagtgctccc ttttccaact cttctcactc 3421 cttcctccat tgtgtttatt acagggccca atgtaaaggc tgttgctact gctgtggaac 3481 agatttaccc atttgtgttt gaaagcagga aagaaatttt agggattcca actactgcaa 3541 gcgagaatct ttattttcag ggcgccgcca aattcaaaga aaccgctgct gctaaattcg 3601 aacgccagca catggacagc ggaggtggag gttcgagcgg tccctcgggt tcgtcgagcc 3661 tggaagttct gttccagggg cccctgtcgt cgagcggtcc ctcgggttcg atggtgagca 3721 aaggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa 3781 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga 3841 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca 3901 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact 3961 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg 4021 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca 4081 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt 4141 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg 4201 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc 4261 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca 4321 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc cttaaggagt 4381 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa tgatctagag 4441 tcgagttaat taagaattcc gccccccccc ctctccctcc ccccccctaa cgttactggc 4501 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg 4561 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct 4621 aggggtcttt cccctctcgc caaaggaatg cagggtctgt tgaatgtcgt gaaggaagca 4681 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg 4741 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct 4801 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa 4861 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt 4921 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa 4981 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa 5041 tatggccaca acctgttaca ttgcacaaga taaaaatata tcatcacgaa cagtaaaact 5101 gtctgcttac ataaacagta atacaagggg tgttatggga tcggccattg aacaagatgg 5161 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca 5221 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt 5281 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg 5341 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga 5401 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca 5461 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct 5521 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac 5581 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc 5641 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt 5701 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt 5761 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg 5821 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat 5881 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgatt 5941 caccacttaa ttggttagaa tctctaactg agcacctttt aaacctgctg cacattggac 6001 tcaaaaggaa aactggacca acaataattg aggaaataga ctcttttatt cattcacggc 6061 tacagtgtaa gctccagtcc ctttggattt tattccaaac cttgctgtaa tataaaagga 6121 agtttacaag acatgatatt gctgctttta caaaaggaca ttctatttat tttcgcagta 6181 attctcatgt ccccataagc agagctgtca cagtgtgcac taccttagat tgttttattg 6241 tcgtcattgt tatttttttc cattttgagc taatgtgttt tatttgtgaa tagtctttta 6301 catttttgta tgctgaatat gggcaccaaa gaacctgtaa aagttatctt tttcaattga 6361 atgtgcacaa ataaaagttt ggaaaagatc tctcttcatt ctataacttt tattccccat 6421 tttctatttt aacaaaaatt gtattcatac ttcttttttt taaaatctat gcaagcttaa 6481 gactctggct aaagtgtgtt ggcttctttt tgaagtgtat tttgtgtgtg tgtgtgtgtg 6541 tgtatatata tatatatatg caccacatgt gtatagtatc ttttaatgct ctgttaccaa 6601 ataacaaata ggaatttttt ttcttgcaga ttagaaataa aaacgtgtat ataaactcta 6661 gggaaccaga ctagaagttt atagataaag gataatgttt tatgttgcta atttaatatg 6721 atagaaaatg ttaaataact tgtaaaggtt aagaaattgt agttttaaga agagaaacct 6781 cttacccact agatggaaat gagaaagaag ctgtgatgaa tggtgcc //