LOCUS Exported 7139 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor TFAP2C with GFP. ACCESSION . VERSION . KEYWORDS pTFAP2C-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7139) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 7139) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7139 /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 3875..3895 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3908..3952 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3992..4015 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 4043..4762 /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(4088..4109) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4349..4368) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4800..5385 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4966..4983) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5193..5212 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5467..6270 /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(5530..5549) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6140..6159 /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 tcggtacccc agtcccgatg gctcaacaaa gaaatattct 2701 tgaaataaat tagggtggag caaaagaaaa tggcaaggga gcttctttta caaagtcaaa 2761 tacattaact ttttgtatga tttagaagag tgcatgactc agccatgagt atttttgagc 2821 tgcttagtta ttcaacccaa agtagttgag ctgccctttc attattgtgt tagcatatct 2881 tacaaaaatg tgttgaaaac ccgatttgtt ttgtcacaac cagtaaagag agattgtgtg 2941 ccttctacct cctgatagca agacttttgt ctttcaaatg taaagatcag ggtggatttg 3001 tgggcaaatt aactgtctcc aaaacgactt aatgcagctg tgggttttct catagaaacc 3061 aacagtaagt tgatttgaaa atgatggtaa atgaagcaga actagtcgag ccaggaaatg 3121 gggtaaaaga gagcgcagtg aaccaggctg tgtcaggagt cagaatttca gggggatgct 3181 tccttctccc ggttcctcac ggaatttgtt tattggaacc aacttgtacc aaggtaaact 3241 gtcccttaag ctcatgggtt ggaatccagg ccccattact tttaccagtc ctggtgacct 3301 tggccagtta gctggggtta agttaatcct gttgtctttt ctaattgcag tgaggattaa 3361 ataagatagt gctttaaaac agcttaactc agagcctggc gcagaatagt tgctaagcgc 3421 agcagttggc tattagtatt aaattctcct tcctcgggtt gaagcagttg tgcttgcctc 3481 ccgggcgcca gcctcagggt agttggttct gtgctcttga cctgtaagga gctagatgga 3541 actcatcgag tcaactgact gtcttcacag gcaactgtgt aaagaattca cagaacttct 3601 cagccaagac cggacacccc atgggaccag caggctcgcc ccagtcttgg agacgaacat 3661 acagaactgc ttgtctcatt tcagtctgat tacccacggg tttggcagcc aggccatctg 3721 tgccgcggtg tctgccctgc agaactacat caaagaagcc ctgattgtca tagacaaatc 3781 ctacatgaac cctggagacc agagtccagc tgattctaac aaaaccctgg agaaaatgga 3841 gaaacacagg aaagggattc caactactgc aagcgagaat ctttattttc agggcgccgc 3901 caaattcaaa gaaaccgctg ctgctaaatt cgaacgccag cacatggaca gcggaggtgg 3961 aggttcgagc ggtccctcgg gttcgtcgag cctggaagtt ctgttccagg ggcccctgtc 4021 gtcgagcggt ccctcgggtt cgatggtgag caaaggcgag gagctgttca ccggggtggt 4081 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga 4141 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa 4201 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag 4261 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta 4321 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt 4381 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 4441 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat 4501 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga 4561 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 4621 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa 4681 cgagaagcgc gatcacatgg tccttaagga gttcgtgacc gccgccggga tcactctcgg 4741 catggacgag ctgtacaagt aatgatctag agtcgagtta attaagaatt ccgccccccc 4801 ccctctccct ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg 4861 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga 4921 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa 4981 tgcagggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa 5041 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct 5101 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg 5161 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg 5221 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca 5281 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg 5341 acgtggtttt cctttgaaaa acacgatgat aatatggcca caacctgtta cattgcacaa 5401 gataaaaata tatcatcacg aacagtaaaa ctgtctgctt acataaacag taatacaagg 5461 ggtgttatgg gatcggccat tgaacaagat ggattgcacg caggttctcc ggccgcttgg 5521 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc 5581 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt 5641 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt 5701 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc 5761 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc 5821 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac 5881 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag 5941 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag 6001 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat 6061 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg 6121 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa 6181 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc 6241 ttctatcgcc ttcttgacga gttcttctga gtcgattgga acgaagaaag gttaggagag 6301 tagggaagga acaggactgc aaaaatcctt ctccaccgca cagactggga acccctcctg 6361 gcctggggga agagtttgtt acctacctta ctatttaaag agccttcact ggttctgcat 6421 cacccgcccc tggacttctt agttgtttct ctagcgctga gctatctcct aactttggac 6481 ctattatcag aaggtgacaa gtactggctc tttattcatt aagctttttt tttttgaacc 6541 ccattctttc cttctctgaa agtggtgcta taagttttag aatcttttaa atacattccc 6601 tgggccaaca gacccacaca cttagccatt gaaatgtcaa attgatgtgc cctagatcaa 6661 cagatcaaca ataccttttt tttcagtgtt aaggtaatgg ttggtttttg tgtccgctaa 6721 atatttacct tgaaaaaaag aaaagtgtgt atctagcttc ttcagagatc aagtcctctg 6781 gtaggaggca aaggttctat ctgcttagca actagttaat aagtggtatc tgacacactc 6841 taaaccccgt gttcaaacgg gggccttctg gttttaggaa acttgtagaa acgaagcctg 6901 ctgattgatt tttttctcct tttttttttt tttttttaac tttgaaagtt aactcttcaa 6961 atgggagact ctttgaaatg acatgttcct ttaaggtact gaagctttat ttgcatattt 7021 atttcagatg tttcgagtaa acttgaaaag ggtaggcacg aagcaatttg ttgctgcttg 7081 tcacccccaa gtccccgtgg aggttctgta ttttaagaaa cagtgcgttg agtgtacag //