LOCUS Exported 6720 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Donor vector for 3' FLAG tag of human BCL6_iso1. ACCESSION . VERSION . KEYWORDS pFETCh_BCL6_iso1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6720) 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 6720) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6720 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 30..47 /label=L4440 /note="L4440 vector, forward primer" protein_bind 347..368 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 383..413 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 421..437 /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 426..448 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 445..461 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 445..461 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 638..840 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 791..811 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 837..861 /label=LNCX /note="Human CMV promoter, forward primer" CDS 937..2067 /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" CDS 3101..3124 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 3128..3151 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 3155..3178 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 3194..3250 /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 3263..4057 /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(3317..3336) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3927..3946 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4578..4595) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4578..4594) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4587..4609) /label=M13/pUC Forward /note="In lacZ gene" CDS complement(4813..5628) /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 5536..5555 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 5620..5639 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 6008..6596 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6497..6516 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagaag 61 ctcgcacatt cagcagcgtt tttcagcgcg ttttcgatca acgtttcaat gttggtatca 121 acaccaggtt taactttgaa cttatcggca ctgacggtta ctgattttga acttttgctt 181 tgccacggaa cggtctgcgt tgtcgggaag atgcgtgatc tgatccttca actcagcaaa 241 agttcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 301 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc 361 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa 421 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg 481 catgcctgca ggtcgactct agaggatccc cgggtaccga gctcgaatta gcttgcatgc 541 ctgcaggtca attccctggc attatgccca gtacatgacc ttatgggact ttcctacttg 601 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat 661 caatgggcgt ggatagcggt ttgactcacg gggatttcaa gtctccaccc cattgacgtc 721 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc 781 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct 841 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccatagg 901 agacaccggg accgatccag cctggggatc ccgaccatgg cttcgtaccc ctgccatcaa 961 cacgcgtctg cgttcgacca ggctgcgcgt tctcgcggcc atagcaaccg acgtacggcg 1021 ttgcgccctc gccggcagca agaagccacg gaagtccgcc tggagcagaa aatgcccacg 1081 ctactgcggg tttatataga cggtcctcac gggatgggga aaaccaccac cacgcaactg 1141 ctggtggccc tgggttcgcg cgacgatatc gtctacgtac ccgagccgat gacttactgg 1201 caggtgctgg gggcttccga gacaatcgcg aacatctaca ccacacaaca ccgcctcgac 1261 cagggtgaga tatcggccgg ggacgcggcg gtggtaatga caagcgccca gataacaatg 1321 ggcatgcctt atgccgtgac cgacgccgtt ctggctcctc atatcggggg ggaggctggg 1381 agctcacatg ccccgccccc ggccctcacc ctcatcttcg accgccatcc catcgccgcc 1441 ctcctgtgct acccggccgc gcgatacctt atgggcagca tgacccccca ggccgtgctg 1501 gcgttcgtgg ccctcatccc gccgaccttg cccggcacaa acatcgtgtt gggggccctt 1561 ccggaggaca gacacatcga ccgcctggcc aaacgccagc gccccggcga gcggcttgac 1621 ctggctatgc tggccgcgat tcgccgcgtt tacgggctgc ttgccaatac ggtgcggtat 1681 ctgcagggcg gcgggtcgtg gcgggaggat tggggacagc tttcggggac ggccgtgccg 1741 ccccagggtg ccgagcccca gagcaacgcg ggcccacgac cccatatcgg ggacacgtta 1801 tttaccctgt ttcgggcccc cgagttgctg gcccccaacg gcgacctgta caacgtgttt 1861 gcctgggcct tggacgtctt ggccaaacgc ctccgtccca tgcacgtctt tatcctggat 1921 tacgaccaat cgcccgccgg ctgccgggac gccctgctgc aacttacctc cgggatggtc 1981 cagacccacg tcaccacccc cggctccata ccgacgatct gcgacctggc gcgcacgttt 2041 gcccgggaga tgggggaggc taactgagga tcccacgtca ctattgtata ctctatatta 2101 tactctatgt tatactctgt aatcctactc aataaacgtg tcacgcctgt gaaaccgtac 2161 taagtctccc gtgtattctt atcaccatca ggtgacatcc tcgcccaggc tgtcaatcat 2221 gccggtatcg attccagtag caccggcccc acgctgacaa cccactcttg cagcgttagc 2281 agcgcccctc ttaacaagcc gacccccacc agcgtcgcgg ttactaacac tcctctcccc 2341 gacctgcagc ccagcttccg tgaattcaga ctgatctagt ccatttgtcc agatgtgttc 2401 agattaattg cataatttga ataagaggta tattctgtga ggtcctagga agattataaa 2461 gtttaggaaa aaaatagcca ctgccctcaa agtgttaccc actgaagaag gatccatgca 2521 cacggagggc tgtgtggtaa aggtagaaat cttacagagg ctcaaaggaa aacaaggatt 2581 attttcatcc ttgagagatt cagaaatgcc ttaatcatta gaagagtttg catctaactg 2641 cacttttgga taagactcaa acttgtggag ttaaggcagg gaaggacgtc tctagcaagg 2701 gagaggcaga agcaacactt gggaaagcag ggaacggaca tttgcacaac gctagcttat 2761 ctgtcggcag aagcccggat gcatatgggt tcttcacctg gccttggaga ggcaagagct 2821 gccccagtcc ctcgtaggga gaatcttcct ctctgggtct ttgagcaatg acatctttct 2881 tgggttgatg acagctaaca gtttgcctct gatatccctg cagtgtgaga agtgtaacct 2941 gcatttccgt cacaaaagcc agctgcgact tcacttgcgc cagaagcatg gcgccatcac 3001 caacaccaag gtgcaatacc gcgtgtcagc cactgacctg cctccggagc tccccaaagc 3061 ctgcgggagc ggaggaggtt ccggtggagg tggttctgga gattacaagg atgacgacga 3121 taagggcgat tacaaggatg acgacgataa gggagattac aaggatgacg acgataaggt 3181 ttcaggaagc ggagctacta acttcagcct gctgaagcag gctggagacg tggaggagaa 3241 ccctggacct ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc 3301 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga 3361 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct 3421 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac 3481 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct 3541 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt 3601 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt 3661 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt 3721 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag 3781 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt 3841 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg 3901 tgtggccgac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg 3961 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg 4021 catcgccttc tatcgccttc ttgacgagtt cttctgattc gaacatcgca tggagtgttg 4081 atgctttcgt ctccagcccc ttctcagaat ctacccaaag gatactgtaa cactttacaa 4141 tgttcatccc atgatgtagt gcctctttca tccactagtg caaatcatag ctgggggttg 4201 ggggtggtgg gggtcggggc ctgggggact gggagccgca gcagctcccc ctcccccact 4261 gccataaaac attaagaaaa tcatattgct tcttctccta tgtgtaaggt gaaccatgtc 4321 agcaaaaagc aaaatcattt tatatgtcaa agcaggggag tatgcaaaag ttctgacttg 4381 actttagtct gcaaaatgag gaatgtatat gttttgtggg aacagatgtt tcttttgtat 4441 gtaaatgtgc attcttttaa aagacaagac ttcagtatgt tgtcaaagag agggctttaa 4501 tttttttaac caaaggtgaa ggaatcttgg aaagtcctct ccactgactg tagcctccaa 4561 ttcactggcc gtcgttcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc 4621 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa 4681 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgcgattta 4741 ttcaacaaag ccgccgtccc gtcaagtcag cgtaatgctc tgccagtgtt acaaccaatt 4801 aaccaattct gattagaaaa actcatcgag catcaaatga aactgcaata tattcatatc 4861 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc 4921 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac 4981 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc 5041 atgagtgacg actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg 5101 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt 5161 cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca 5221 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc 5281 tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag 5341 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc 5401 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc 5461 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc 5521 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga 5581 atttaatcgc ggcttcgagc aagacgtttc ccgttgaata tggctcataa caccccttgt 5641 attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc 5701 aatgtaacat cagagatttt gagacacaac gtggctttgt tgaataaatc gaacttttgc 5761 tgagttgaag gatcagatca cgcatcttcc cgacaacgca gaccgttccg tggcaaagca 5821 aaagttcaaa atcaccaact ggtccaccta caacaaagct ctcatcaacc gtggctccct 5881 cactttctgg ctggatgatg gggcgattca ggcctggtat gagtcagcaa caccttcttc 5941 acgaggcaga cctctcgacg gagttccact gagcgtcaga ccccgtagaa aagatcaaag 6001 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 6061 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 6121 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc 6181 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 6241 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 6301 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 6361 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc 6421 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 6481 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 6541 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 6601 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct 6661 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata //