LOCUS Exported 6970 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Donor vector for 3' FLAG tag of human NR2F6. ACCESSION . VERSION . KEYWORDS pFETCh_NR2F6 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6970) 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 6970) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6970 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 763..786 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 790..813 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 817..840 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 856..912 /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 925..1719 /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(979..998) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 1589..1608 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(2490..2507) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2490..2506) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2499..2521) /label=M13/pUC Forward /note="In lacZ gene" CDS complement(2725..3540) /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 ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNILQFHLMLDEFF" primer_bind 3448..3467 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 3532..3551 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 3920..4508 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4409..4428 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4662..4679 /label=L4440 /note="L4440 vector, forward primer" protein_bind 4979..5000 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5015..5045 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5053..5069 /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 5058..5080 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5077..5093 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5077..5093 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 5270..5472 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5423..5443 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 5469..5493 /label=LNCX /note="Human CMV promoter, forward primer" CDS 5569..6699 /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 ccgacctgca gcccagctcg atcctcccat tagccttccg agtagctgga actgcaggca 61 cgcactacca tgcctggcta attattctta tttttagtag agacggaata tcatcatgtt 121 ggccaggctg gtcttgaact cctgacctca agtgatccac ctgcctcagc ctcccaaagt 181 gctgggatta caggcgtgag ctactgtgcc cagccccagc tgatttgttt ttgcagagaa 241 gggatcatac tatgttgccc tggctggtct caaatcctgg ccttaaatga tgcttctgcc 301 ttggcctccc aaagtgctgg gactgtggcc gtgagccatc atgcccggcc ccctgttctt 361 acccacagtg gcgagtggtg tcccctagtc acccctgtgg gctctgtgtt ctctagctgc 421 ctcccacctg tctgacttgc ctttgtcccc ctagacgcct gtggcctctc agacccggcc 481 cacgttgaga gcctgcagga gaaggcgcag gtggccctca ccgagtatgt gcgggcgcag 541 tacccgtccc agccccagcg cttcgggcgc ctgctgctgc ggctccccgc cctgcgcgcg 601 gtccctgcct ccctcatctc ccagctgttc ttcatgcgcc tggtggggaa gacgcccatt 661 gagacactga tcagagacat gctgctgtcg gggagtacct tcaactggcc ctacggctcg 721 ggccagggga gcggaggagg ttccggtgga ggtggttctg gagattacaa ggatgacgac 781 gataagggcg attacaagga tgacgacgat aagggagatt acaaggatga cgacgataag 841 gtttcaggaa gcggagctac taacttcagc ctgctgaagc aggctggaga cgtggaggag 901 aaccctggac ctggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg 961 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 1021 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 1081 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg 1141 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 1201 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 1261 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 1321 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 1381 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 1441 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc 1501 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 1561 ggtgtggccg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 1621 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 1681 cgcatcgcct tctatcgcct tcttgacgag ttcttctgat tcgaacatca cggggccacg 1741 tgtgctgtgg ccaggcctgc agacagacct caagggacag ggaatgctga ggcctcgagg 1801 ggcctcccgg ggcccaggac tctggcttct ctcctcagac ttctattttt taaagactgt 1861 gaaatgtttg tcttttctgt tttttaaatg atcatgaaac caaaaagaga ctgatcatcc 1921 aggcctcagc ctcatcctcc ccaggacccc tgtccaggat ggagggtcca attctaggac 1981 agccttgttc ctcagcaccc ctagcatgaa cttgtgggat ggtggggttg gcttccctgg 2041 catgatggac aaaggcctgg cgtcggccag aggggctgct ccagtgggca ggggtagcta 2101 gcgtgtgcca ggcagatcct ctggacacgt aacctatgtc agacactaca tgatgactca 2161 aggccaataa taaagacatt tcctacctgc acagcctccc tggtctttat ttttattttt 2221 tatttttgag acggagtcct acttcgtcac ccaggctgga gtgcagtggc gtgatcttgg 2281 ctcactgtaa cctctgcctc acaggttcaa gcgattctcc tgcctcagcc tccccagtag 2341 ctggaactac aggcacccac cacgatgccc agctaatttt atttttggta gagatggggt 2401 ttcaccatgt tgaccaggtt ggtctcgaac tcctgacctt ggaaagtcct ctccactgac 2461 tgtagcctcc aattcactgg ccgtcgttca ctggccgtcg ttttacaacg tcgtgactgg 2521 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg 2581 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc 2641 gaatgcgatt tattcaacaa agccgccgtc ccgtcaagtc agcgtaatgc tctgccagtg 2701 ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa 2761 tatattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg 2821 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc 2881 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag 2941 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc 3001 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac 3061 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa 3121 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac 3181 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat 3241 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag 3301 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac 3361 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata 3421 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc 3481 atccatgttg gaatttaatc gcggcttcga gcaagacgtt tcccgttgaa tatggctcat 3541 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt 3601 tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa 3661 tcgaactttt gctgagttga aggatcagat cacgcatctt cccgacaacg cagaccgttc 3721 cgtggcaaag caaaagttca aaatcaccaa ctggtccacc tacaacaaag ctctcatcaa 3781 ccgtggctcc ctcactttct ggctggatga tggggcgatt caggcctggt atgagtcagc 3841 aacaccttct tcacgaggca gacctctcga cggagttcca ctgagcgtca gaccccgtag 3901 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 3961 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 4021 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc 4081 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 4141 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 4201 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc 4261 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa 4321 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 4381 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 4441 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 4501 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg 4561 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg 4621 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg 4681 aagcggaaga agctcgcaca ttcagcagcg tttttcagcg cgttttcgat caacgtttca 4741 atgttggtat caacaccagg tttaactttg aacttatcgg cactgacggt tactgatttt 4801 gaacttttgc tttgccacgg aacggtctgc gttgtcggga agatgcgtga tctgatcctt 4861 caactcagca aaagttccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 4921 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta 4981 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta 5041 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt 5101 acgccaagct tgcatgcctg caggtcgact ctagaggatc cccgggtacc gagctcgaat 5161 tagcttgcat gcctgcaggt caattccctg gcattatgcc cagtacatga ccttatggga 5221 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt 5281 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc aagtctccac 5341 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt 5401 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat 5461 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt 5521 gacctccata ggagacaccg ggaccgatcc agcctgggga tcccgaccat ggcttcgtac 5581 ccctgccatc aacacgcgtc tgcgttcgac caggctgcgc gttctcgcgg ccatagcaac 5641 cgacgtacgg cgttgcgccc tcgccggcag caagaagcca cggaagtccg cctggagcag 5701 aaaatgccca cgctactgcg ggtttatata gacggtcctc acgggatggg gaaaaccacc 5761 accacgcaac tgctggtggc cctgggttcg cgcgacgata tcgtctacgt acccgagccg 5821 atgacttact ggcaggtgct gggggcttcc gagacaatcg cgaacatcta caccacacaa 5881 caccgcctcg accagggtga gatatcggcc ggggacgcgg cggtggtaat gacaagcgcc 5941 cagataacaa tgggcatgcc ttatgccgtg accgacgccg ttctggctcc tcatatcggg 6001 ggggaggctg ggagctcaca tgccccgccc ccggccctca ccctcatctt cgaccgccat 6061 cccatcgccg ccctcctgtg ctacccggcc gcgcgatacc ttatgggcag catgaccccc 6121 caggccgtgc tggcgttcgt ggccctcatc ccgccgacct tgcccggcac aaacatcgtg 6181 ttgggggccc ttccggagga cagacacatc gaccgcctgg ccaaacgcca gcgccccggc 6241 gagcggcttg acctggctat gctggccgcg attcgccgcg tttacgggct gcttgccaat 6301 acggtgcggt atctgcaggg cggcgggtcg tggcgggagg attggggaca gctttcgggg 6361 acggccgtgc cgccccaggg tgccgagccc cagagcaacg cgggcccacg accccatatc 6421 ggggacacgt tatttaccct gtttcgggcc cccgagttgc tggcccccaa cggcgacctg 6481 tacaacgtgt ttgcctgggc cttggacgtc ttggccaaac gcctccgtcc catgcacgtc 6541 tttatcctgg attacgacca atcgcccgcc ggctgccggg acgccctgct gcaacttacc 6601 tccgggatgg tccagaccca cgtcaccacc cccggctcca taccgacgat ctgcgacctg 6661 gcgcgcacgt ttgcccggga gatgggggag gctaactgag gatcccacgt cactattgta 6721 tactctatat tatactctat gttatactct gtaatcctac tcaataaacg tgtcacgcct 6781 gtgaaaccgt actaagtctc ccgtgtattc ttatcaccat caggtgacat cctcgcccag 6841 gctgtcaatc atgccggtat cgattccagt agcaccggcc ccacgctgac aacccactct 6901 tgcagcgtta gcagcgcccc tcttaacaag ccgaccccca ccagcgtcgc ggttactaac 6961 actcctctcc //