LOCUS Exported 8043 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor JUNB with GFP. ACCESSION . VERSION . KEYWORDS pJUNB-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8043) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 8043) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8043 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(70..89) /label=T7 /note="T7 promoter, forward primer" promoter complement(71..89) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(96..113) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(96..112) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(105..127) /label=M13/pUC Forward /note="In lacZ gene" CDS 250..552 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 431..450 /label=ccdB-fwd /note="ccdB gene, forward primer" CDS 901..1695 /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" regulatory 1895..1904 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1901..2275 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" rep_origin 2413..3001 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2902..2921 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3155..3172 /label=L4440 /note="L4440 vector, forward primer" protein_bind 3289..3310 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 3325..3355 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3363..3379 /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 3368..3390 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 3387..3403 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 3387..3403 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 3421..3439 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 3421..3438 /label=SP6 /note="SP6 promoter, forward primer" CDS 4581..4601 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 4614..4658 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 4698..4721 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 4749..5468 /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(4794..4815) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(5055..5074) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 5506..6091 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(5672..5689) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5899..5918 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 6173..6976 /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="MGSAIGQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" ORIGIN 1 aagggcgaat tctgcagata tccatcacac tggcggccgc tcgagcatgc atctagaggg 61 cccaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt 121 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc 181 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagccta 241 tacgtacggc agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg 301 gatgtacaga gtgatattat tgacacgccg gggcgacgga tggtgatccc cctggccagt 361 gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat 421 gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa 481 gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc 541 tggggaatat aaatgtcagg catgagatta tcaaaaagga tcttcaccta gatccttttc 601 acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag ctactgggct 661 atctggacaa gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag tgggcttaca 721 tggcgatagc tagactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg 781 gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca 841 aggatctgat ggcgcagggg atcaagctct gatcaagaga caggatgagg atcgtttcgc 901 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc 961 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca 1021 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg 1081 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg 1141 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag 1201 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg 1261 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc 1321 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa 1381 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac 1441 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat 1501 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac 1561 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc 1621 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt 1681 gacgagttct tctgaattat taacgcttac aatttcctga tgcggtattt tctccttacg 1741 catctgtgcg gtatttcaca ccgcatcagg tggcactttt cggggaaatg tgcgcggaac 1801 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc 1861 ctgataaatg cttcaataat agcacgtgag gagggccacc atggccaagt tgaccagtgc 1921 cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct 1981 cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac 2041 cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg 2101 ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg 2161 ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc 2221 cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt 2281 gctaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat 2341 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat 2401 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa 2461 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa 2521 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt 2581 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt 2641 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata 2701 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt 2761 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac 2821 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga 2881 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg 2941 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa 3001 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat 3061 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc 3121 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga 3181 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg 3241 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta 3301 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg 3361 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct 3421 atttaggtga cactatagaa tactcaagct atgcatcaag cttggtaccg agctcggatc 3481 cactagtaac ggccgccagt gtgctggata tctgcagaat tcgcccttaa cagcccttct 3541 accacgacga ctcatacaca gctacgggat acggccgggc ccctggtggc ctctctctac 3601 acgactacaa actcctgaaa ccgagcctgg cggtcaacct ggccgacccc taccggagtc 3661 tcaaagcgcc tggggctcgc ggacccggcc cagagggcgg cggtggcggc agctactttt 3721 ctggtcaggg ctcggacacc ggcgcgtctc tcaagctcgc ctcttcggag ctggaacgcc 3781 tgattgtccc caacagcaac ggcgtgatca cgacgacgcc tacacccccg ggacagtact 3841 tttacccccg cgggggtggc agcggtggag gtgcaggggg cgcagggggc ggcgtcaccg 3901 aggagcagga gggcttcgcc gacggctttg tcaaagccct ggacgatctg cacaagatga 3961 accacgtgac accccccaac gtgtccctgg gcgctaccgg ggggcccccg gctgggcccg 4021 ggggcgtcta cgccggcccg gagccacctc ccgtttacac caacctcagc agctactccc 4081 cagcctctgc gtcctcggga ggcgccgggg ctgccgtcgg gaccgggagc tcgtacccga 4141 cgaccaccat cagctacctc ccacacgcgc cgcccttcgc cggtggccac ccggcgcagc 4201 tgggcttggg ccgcggcgcc tccaccttca aggaggaacc gcagaccgtg ccggaggcgc 4261 gcagccggga cgccacgccg ccggtgtccc ccatcaacat ggaagaccaa gagcgcatca 4321 aagtggagcg caagcggctg cggaaccggc tggcggccac caagtgccgg aagcggaagc 4381 tggagcgcat cgcgcgcctg gaggacaagg tgaagacgct caaggccgag aacgcggggc 4441 tgtcgagtac cgccggcctc ctccgggagc aggtggccca gctcaaacag aaggtcatga 4501 cccacgtcag caacggctgt cagctgctgc ttggggtcaa gggacacgcc ttcgattatg 4561 atattccaac tactgcaagc gagaatcttt attttcaggg cgccgccaaa ttcaaagaaa 4621 ccgctgctgc taaattcgaa cgccagcaca tggacagcgg aggtggaggt tcgagcggtc 4681 cctcgggttc gtcgagcctg gaagttctgt tccaggggcc cctgtcgtcg agcggtccct 4741 cgggttcgat ggtgagcaaa ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg 4801 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg 4861 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct 4921 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc 4981 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca 5041 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg 5101 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc 5161 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc 5221 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc 5281 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg 5341 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc 5401 acatggtcct taaggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt 5461 acaagtaatg atctagagtc gagttaatta agaattccgc ccccccccct ctccctcccc 5521 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg 5581 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc 5641 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca gggtctgttg 5701 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg 5761 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca 5821 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata 5881 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc 5941 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt 6001 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt 6061 tgaaaaacac gatgataata tggccacaac ctgttacatt gcacaagata aaaatatatc 6121 atcacgaaca gtaaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgggatc 6181 ggccattgga caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt 6241 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc 6301 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact 6361 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt 6421 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca 6481 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat 6541 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg 6601 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga 6661 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga 6721 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa 6781 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga 6841 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt 6901 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct 6961 tgacgagttc ttctgaaacg tcccctgccc ctttacggac accccctcgc ttggacggct 7021 gggcacacgc ctcccactgg ggtccaggga gcaggcggtg ggcacccacc ctgggaccta 7081 ggggcgccgc aaaccacact ggactccggc cctcctaccc tgcgcccagt ccttccacct 7141 cgacgtttac aagccccccc ttccactttt ttttgtatgt tttttttctg ctggaaacag 7201 actcgattca tattgaatat aatatatttg tgtatttaac agggagggga agagggggcg 7261 atcgcggcgg agctggcccc gccgcctggt actcaagccc gcggggacat tgggaagggg 7321 acccccgccc cctgccctcc cctctctgca ccgtactgtg gaaaagaaac acgcacttag 7381 tctctaaaga gtttatttta agacgtgttt gtgtttgtgt gtgtttgttc tttttattga 7441 atctatttaa gtaaaaaaaa aattggttct ttattaattt ctgttgtctt tttttccaag 7501 ctgggagggc ggggggaaaa aaaaaagcac tggtttgccc ccagctcagt gctgttggtg 7561 gctcggtcct gtatgtgtcc ccctcgtcgg ttcggcgcag gcatcttgtg gtcccagccc 7621 aggagtccca cccttcccgc gtccccagat ctccagggtt ggatggttgg ggcgcgggac 7681 gcggtccagg gacccaggag ctgaaggcag ggtgctccgg ccgagacttg gagtgcgcag 7741 gcgcgtcccc gcccagccgc gcgcgccggg gctttccccg ctgacgcagc ggaagcgctg 7801 cccatacaag gaccgattct gcccagtgac gcgaccgcgg tctctgggca gattccggga 7861 atcccctccc ccgctctgcc gggcagggcc gcggcgccgg gaagggggcc gggattttcc 7921 cgggcaggcg gctgcccggg cacggaaagc cccaggcctg ggtccagagc gcccgcggtg 7981 ggcgggcagc gctcctgggc tccgggagcc acccctgtgc caccttcaag acactggcgc 8041 cta //