LOCUS Exported 7033 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor OVOL1 with GFP. ACCESSION . VERSION . KEYWORDS pOVOL1-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7033) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 7033) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7033 /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 3564..3584 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3597..3641 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3681..3704 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 3732..4451 /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(3777..3798) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4038..4057) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4489..5074 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4655..4672) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4882..4901 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5156..5959 /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(5219..5238) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5829..5848 /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 cgtcttccca atctgccagc ctgtggcccc 2701 aacttctgga tgtctgtcag caatgccgtc ctcacccaca ggtgaccctt ggggacagtc 2761 ccagtggaga cctgttcacc tgccgtgtct gccagaaggc cttcacctac cagcgcatgc 2821 tgaaccgcca catgaagtgt cacaacgacg tcaagaggca cctctgcacg tactgcggga 2881 agggcttcaa tgacaccttc gacctcaaga gacacgtccg aactcacact ggtaagtgga 2941 agcccacggc tgggaggagc acgccggatc cgcctggccg cggccgtgcg ggaaaagtcc 3001 agcaccccct gctctccctc ggtgacgggc agcctgggag ggcatcccgg agctcagggc 3061 caaagtcctc ctgagctcat cagttaggag ttttgatttg agagtcactc gggaagttgg 3121 tgttggacag gcctccgtcc atcccttggc cctagaggca ggggtcctgt cctttctgtg 3181 tctggaagca gccgacagcc tctgaagtcc ctgccaggtc tctgatgctg gcccctgtcc 3241 tccccacagg cgtgcggccc tacaagtgca gcctgtgtga caaggccttc acgcagcgct 3301 gctctctgga gtctcacctc aagaagatcc atggtgtgca gcagaagtac gcgtacaagg 3361 agcggcgggc caagctgtac gtgtgtgagg agtgcggctg cacatctgag agccaggagg 3421 gccacgtcct gcacctgaag gagcaccacc ctgacagccc gctgctgcgc aagacctcca 3481 agaaggtggc cgtggcacta cagaacactg tcacttccct gctgcagggc agcccccacc 3541 tggggattcc aactactgca agagagaatc tttattttca gggcgccgcc aaattcaaag 3601 aaaccgctgc tgctaaattc gaacgccagc acatggacag cggaggtgga ggttcgagcg 3661 gtccctcggg ttcgtcgagc ctggaagttc tgttccaggg gcccctgtcg tcgagcggtc 3721 cctcgggttc gatggtgagc aaaggcgagg agctgttcac cggggtggtg cccatcctgg 3781 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg 3841 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc 3901 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg 3961 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc 4021 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg 4081 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca 4141 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca 4201 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg 4261 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc 4321 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg 4381 atcacatggt ccttaaggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc 4441 tgtacaagta atgatctaga gtcgagttaa ttaagaattc cgcccccccc cctctccctc 4501 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat 4561 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct 4621 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcagggtctg 4681 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta 4741 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag 4801 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg 4861 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat 4921 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca 4981 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc 5041 ctttgaaaaa cacgatgata atatggccac aacctgttac attgcacaag ataaaaatat 5101 atcatcacga acagtaaaac tgtctgctta cataaacagt aatacaaggg gtgttatggg 5161 atcggccatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct 5221 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct 5281 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga 5341 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc 5401 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg 5461 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc 5521 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca 5581 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga 5641 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc 5701 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga 5761 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca 5821 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg 5881 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct 5941 tcttgacgag ttcttctgag tcgtggctcg agccctgggg gtgctcctgg aagccccaag 6001 agcatccagg attgcctccc agctgcctgg ccagcccacc ctcctgcaac ctctcacccg 6061 aacaccagtg atcaggactg gagcccccgt gccttggtct cccccctggg cacacgtgct 6121 cactcaggcc cagcaatgac ctctgctcat ttttgcattt ttgacttatg ggccgaggct 6181 gttctgagcc tgggaagatg tacctatgtc aagagaaggg atgaggccaa ggctgccttc 6241 aattagaagc agccgcccac agagacaggc actgtgtgcc tggcagcagg acttcctacc 6301 cagaggaggt tcgagctagg atcccactgc ccccgcctct cagcacaggg caggggctgc 6361 aggtccccag tggacatcag agtcaaaagc actggcaaag ggtacccctg caaacaactg 6421 tggtgggggc tggcagcaga ccccccacct ggcagggctt ctaatgctca gggttctgga 6481 gggctctgtc cttccggcaa ggagaggcac acatgtgtgc ccagccgtgt gtgtgcgtgt 6541 gcttgtgtgt gtgcactgct gtgtgtgtgt gcacgcacag gaagcctttc cacatatcac 6601 ctcatttcta agaaataaac tacaaggtgc caagaaggtt ttatttcctt ttttttttta 6661 aagatgacaa atgtacagat gttaatatat ttttggtgcc aatggcgatg tttttaagag 6721 tgggatggag ctggcttttc tccattcccg tgcgcttcta tttatcctgg acatttcaaa 6781 cctcctctgt gccttggctc tgggcggggg ctgccccaac ccacccccgt tctttgtacg 6841 tgctgagaca gccactagaa gatcttcctc cagcggcgcc ctggacggct gctcctgcga 6901 acagcccatg gcatcttctg ctcttccctc ccggctctgc cctgcacatc ctgttgagcc 6961 aagccccagt gacccggaga gctggcctga tgctgagagt gtgtcctcct ggggctttag 7021 ggggcaggaa ggt //