LOCUS Exported 7154 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Donor vector for 3' FLAG tag of human TEAD3. ACCESSION . VERSION . KEYWORDS pFETCh_TEAD3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7154) AUTHORS Savic D, Partridge EC, Newberry KM, Smith SB, Meadows SK, Roberts BS, Mackiewicz M, Mendenhall EM, Myers RM TITLE CETCh-seq: CRISPR epitope tagging ChIP-seq of DNA-binding proteins. JOURNAL Genome Res. 2015 Sep 9. PUBMED 26355004 REFERENCE 2 (bases 1 to 7154) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7154 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 837..860 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 864..887 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 891..914 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 930..986 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 999..1793 /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(1053..1072) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 1663..1682 /label=Neo-F /note="Neomycin resistance gene, forward primer" CDS 2516..2527 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(2656..2673) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2656..2672) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2665..2687) /label=M13/pUC Forward /note="In lacZ gene" CDS complement(2891..3706) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 3614..3633 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 3698..3717 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 4086..4674 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4575..4594 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4828..4845 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5145..5166 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5181..5211 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5219..5235 /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 5224..5246 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5243..5259 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5243..5259 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 5436..5638 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5589..5609 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 5635..5659 /label=LNCX /note="Human CMV promoter, forward primer" CDS 5735..6865 /codon_start=1 /product="herpes simplex virus thymidine kinase" /label=HSV TK /note="confers sensitivity to nucleoside analogs such as acyclovir or ganciclovir" /translation="MASYPCHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRLEQK MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWQVLGASETIANIYTTQ HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR PGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAVPPQGAEPQSNAGPR PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDAL LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN" ORIGIN 1 attgacctct gaccctgctg catcctcatg gcttgtgggc agagtgccag agatggatcc 61 ttgggcagga ctctgatgtg gtgtggcggg aatacttctg gtctctgcca gcacctaaca 121 ccccccaccc ccatgcccag gccgacctca acagcaccat ccaggagggc ccgggagcct 181 tctatggggt cagctctcag tacagctctg ctgatagcat gaccatcagc gtctccacca 241 aggtgtgctc ctttggcaaa caggtggtag agaaggtgga ggtgagtggg ggctctgggt 301 ggcttggtgg ggctgggagc aggcaggccg agtgacgcct tctccatatt ctctgcagac 361 tgagtatgcc aggctggaga acgggcgctt tgtgtaccgt atccaccgct cgcccatgtg 421 cgagtacatg atcaacttca tccacaagct gaagcacctg cccgagaagt acatgatgaa 481 cagcgtgctg gagaacttca ccatcctgca ggtatggggg cagcaggggt cgtggggagt 541 cccacaggcc agggcctcca agttgggtag ccagacacat cgcttggctg agaggcatag 601 atccggaaag gagaatggtc tgttcagccc cctgccccgt ggcccttttc tccctgacct 661 cctgaatctc ttacctgctc accccccagg tggtcacgag ccgggactcc caggagaccc 721 tgcttgtcat tgcttttgtc ttcgaagtct ccaccagtga gcacggggcc cagcaccatg 781 tctacaagct cgtcaaagat gggagcggag gaggttccgg tggaggtggt tctggagatt 841 acaaggatga cgacgataag ggcgattaca aggatgacga cgataaggga gattacaagg 901 atgacgacga taaggtttca ggaagcggag ctactaactt cagcctgctg aagcaggctg 961 gagacgtgga ggagaaccct ggacctggat cgtttcgcat gattgaacaa gatggattgc 1021 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga 1081 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt 1141 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat 1201 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg 1261 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg 1321 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc 1381 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga 1441 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag 1501 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc 1561 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg 1621 actgtggccg gctgggtgtg gccgaccgct atcaggacat agcgttggct acccgtgata 1681 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg 1741 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgattcgaac 1801 atccgcctcc ttaaggatgc agggtgagca tctcctctcc acacctgcct ggcacccctg 1861 ggggggtcca ggattgagga ttcatctacc tgccaggcct caggcccagg acccaggagg 1921 cctccccacc taccccagca cacacactcc ctgccactgt tctgcgcttt aattgtggga 1981 gaagagagga gaggagggct cagcggtggg gcagcctgtc cggggcgctg acccaccatc 2041 accctgctct gcccagcctc gcgtgacctc agagaggtgg ggatagggga caccttcagc 2101 ctccagcatg tgtggccact gtacccccac ccacccttgg gggagcatga tgggcaggtg 2161 agggcaggat ggagaccaag ggagtcagtg agcagaggcc ctgggagtgt ccggttgggg 2221 ttggactgag gacagagggg cccacacttc cttgcccctt tgtgtcccag gcctggtgcc 2281 cagactcctt gcatggcttg tgtggtcctc agactccgca cagcgagcgt aggtctctgg 2341 gtttcagatg aagtgcccag gctccaggaa gttgagggac ccacaggaga ggtgggcaga 2401 gctggagttc tcatccaggg ctgcttgtcc ccagagccca ggtttatact acctccctgg 2461 ggcgggggct ggccgcaggg taggggagag gctctgcagt gtggagtgga gcctcatcga 2521 ggggcgctgg gttaggggag cacctgtttc agactgggca tgaagaaggg agcacagcag 2581 ctactagacc ccattagcac ctccttggaa agtcctctcc actgactgta gcctccaatt 2641 cactggccgt cgttcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt 2701 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga 2761 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gcgatttatt 2821 caacaaagcc gccgtcccgt caagtcagcg taatgctctg ccagtgttac aaccaattaa 2881 ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag 2941 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga 3001 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat 3061 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat 3121 gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt 3181 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca 3241 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa 3301 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg 3361 aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta 3421 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg 3481 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat 3541 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg 3601 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat 3661 ttaatcgcgg cttcgagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat 3721 tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa 3781 tgtaacatca gagattttga gacacaacgt ggctttgttg aataaatcga acttttgctg 3841 agttgaagga tcagatcacg catcttcccg acaacgcaga ccgttccgtg gcaaagcaaa 3901 agttcaaaat caccaactgg tccacctaca acaaagctct catcaaccgt ggctccctca 3961 ctttctggct ggatgatggg gcgattcagg cctggtatga gtcagcaaca ccttcttcac 4021 gaggcagacc tctcgacgga gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 4081 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 4141 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact 4201 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac 4261 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 4321 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 4381 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 4441 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc 4501 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 4561 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 4621 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc 4681 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt 4741 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc 4801 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagaagct 4861 cgcacattca gcagcgtttt tcagcgcgtt ttcgatcaac gtttcaatgt tggtatcaac 4921 accaggttta actttgaact tatcggcact gacggttact gattttgaac ttttgctttg 4981 ccacggaacg gtctgcgttg tcgggaagat gcgtgatctg atccttcaac tcagcaaaag 5041 ttcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac 5101 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc 5161 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt 5221 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgca 5281 tgcctgcagg tcgactctag aggatccccg ggtaccgagc tcgaattagc ttgcatgcct 5341 gcaggtcaat tccctggcat tatgcccagt acatgacctt atgggacttt cctacttggc 5401 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca 5461 atgggcgtgg atagcggttt gactcacggg gatttcaagt ctccacccca ttgacgtcaa 5521 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc 5581 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg 5641 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccataggag 5701 acaccgggac cgatccagcc tggggatccc gaccatggct tcgtacccct gccatcaaca 5761 cgcgtctgcg ttcgaccagg ctgcgcgttc tcgcggccat agcaaccgac gtacggcgtt 5821 gcgccctcgc cggcagcaag aagccacgga agtccgcctg gagcagaaaa tgcccacgct 5881 actgcgggtt tatatagacg gtcctcacgg gatggggaaa accaccacca cgcaactgct 5941 ggtggccctg ggttcgcgcg acgatatcgt ctacgtaccc gagccgatga cttactggca 6001 ggtgctgggg gcttccgaga caatcgcgaa catctacacc acacaacacc gcctcgacca 6061 gggtgagata tcggccgggg acgcggcggt ggtaatgaca agcgcccaga taacaatggg 6121 catgccttat gccgtgaccg acgccgttct ggctcctcat atcggggggg aggctgggag 6181 ctcacatgcc ccgcccccgg ccctcaccct catcttcgac cgccatccca tcgccgccct 6241 cctgtgctac ccggccgcgc gataccttat gggcagcatg accccccagg ccgtgctggc 6301 gttcgtggcc ctcatcccgc cgaccttgcc cggcacaaac atcgtgttgg gggcccttcc 6361 ggaggacaga cacatcgacc gcctggccaa acgccagcgc cccggcgagc ggcttgacct 6421 ggctatgctg gccgcgattc gccgcgttta cgggctgctt gccaatacgg tgcggtatct 6481 gcagggcggc gggtcgtggc gggaggattg gggacagctt tcggggacgg ccgtgccgcc 6541 ccagggtgcc gagccccaga gcaacgcggg cccacgaccc catatcgggg acacgttatt 6601 taccctgttt cgggcccccg agttgctggc ccccaacggc gacctgtaca acgtgtttgc 6661 ctgggccttg gacgtcttgg ccaaacgcct ccgtcccatg cacgtcttta tcctggatta 6721 cgaccaatcg cccgccggct gccgggacgc cctgctgcaa cttacctccg ggatggtcca 6781 gacccacgtc accacccccg gctccatacc gacgatctgc gacctggcgc gcacgtttgc 6841 ccgggagatg ggggaggcta actgaggatc ccacgtcact attgtatact ctatattata 6901 ctctatgtta tactctgtaa tcctactcaa taaacgtgtc acgcctgtga aaccgtacta 6961 agtctcccgt gtattcttat caccatcagg tgacatcctc gcccaggctg tcaatcatgc 7021 cggtatcgat tccagtagca ccggccccac gctgacaacc cactcttgca gcgttagcag 7081 cgcccctctt aacaagccga cccccaccag cgtcgcggtt actaacactc ctctccccga 7141 cctgcagccc agct //