LOCUS Exported 6967 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Donor vector for 3' FLAG tag of human RERE. ACCESSION . VERSION . KEYWORDS pFETCh_RERE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6967) 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 6967) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6967 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(216..233) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(387..975) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(467..486) /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind complement(1344..1363) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1355..2170 /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 complement(1428..1447) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 2374..2396 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2388..2405 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2389..2405 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS complement(3176..3970) /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(3287..3306) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 3897..3916 /label=Neo-R /note="Neomycin resistance gene, reverse primer" CDS complement(3983..4039) /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 complement(4055..4078) /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS complement(4082..4105) /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS complement(4109..4132) /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS complement(5163..6293) /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" primer_bind complement(6369..6393) /label=LNCX /note="Human CMV promoter, forward primer" promoter complement(6390..6592) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(6419..6439) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind complement(6769..6785) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6769..6785) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6782..6804) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6793..6809 /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(6817..6847) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6862..6883 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 ggcggtttgc gtattgggaa cttttgctga gttgaaggat cagatcacgc atcttcccga 61 caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc agtaaccgtc agtgccgata 121 agttcaaagt taaacctggt gttgatacca acattgaaac gttgatcgaa aacgcgctga 181 aaaacgctgc tgaatgtgcg agcttcttcc gcttcctcgc tcactgactc gctgcgctcg 241 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 301 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 361 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 421 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 481 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 541 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat 601 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 661 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 721 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 781 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt 841 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 901 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 961 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 1021 tccgtcgaga ggtctgcctc gtgaagaagg tgttgctgac tcataccagg cctgaatcgc 1081 cccatcatcc agccagaaag tgagggagcc acggttgatg agagctttgt tgtaggtgga 1141 ccagttggtg attttgaact tttgctttgc cacggaacgg tctgcgttgt cgggaagatg 1201 cgtgatctga tccttcaact cagcaaaagt tcgatttatt caacaaagcc acgttgtgtc 1261 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact 1321 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc 1381 ttgctcgaag ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc 1441 tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc 1501 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat 1561 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg 1621 tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt 1681 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg 1741 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct 1801 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga 1861 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataagcttt tgccattctc 1921 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg 1981 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct 2041 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca 2101 aaaatatggt attgataatc ctgatatgaa tatattgcag tttcatttga tgctcgatga 2161 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg 2221 acgggacggc ggctttgttg aataaatcgc attcgccatt caggctgcgc aactgttggg 2281 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg 2341 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg 2401 ccagtgaacg acggccagtg aattggaggc tacagtcagt ggagaggact ttccaagtct 2461 gtccatgtgg aacgcccctt tgctcccttt aattaaaggg ataattatat ataacttttt 2521 attaaaaaat caactctgta ttctgtcaat agttggtagg aattcacaat tagtgacatg 2581 gctggaaaaa atagatcaca atagtagttt atggaagaaa aaaaggctgt tctaaaatta 2641 tttatttaca gacgtaaaaa gccatgctgc agaactttct ccaattatga agatttccta 2701 caatgtatta aaataaaaga tttttttttt taaaaattat agctcttgga ttccaaattc 2761 ctatgccccc gatctgcaag cctcgtgggc tctggagcac tcgtggcgga gtgtgtcagt 2821 gtgtctgcac gcacagtgaa gggtgtgaac tggattctag cacctactga gaaatcaagg 2881 aaaagaaaac taaagccaaa ccccaagatt gcagggaagc tttctggatg ttcagtccag 2941 tccaggactc actaagtttc tgggggactg tcatgcaggg agatccaaac cagtctcagg 3001 aaatcctctc gacaaacgaa cactactatg tggatacatt tttagttgtg ggtttttaaa 3061 tatataaaga aatctttaga agatattctt tttgcttttg cagctcctat tttatgtaaa 3121 aagtcctgtt tctcccccca agaactgggg tttccacagc cagcggatgt tcgaatcaga 3181 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt 3241 aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag 3301 ccaacgctat gtcctgatag cggtcggcca cacccagccg gccacagtcg atgaatccag 3361 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga 3421 gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc 3481 cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg 3541 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat 3601 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg 3661 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga 3721 caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg 3781 cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc 3841 gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc 3901 agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt 3961 gttcaatcat gcgaaacgat ccaggtccag ggttctcctc cacgtctcca gcctgcttca 4021 gcaggctgaa gttagtagct ccgcttcctg aaaccttatc gtcgtcatcc ttgtaatctc 4081 ccttatcgtc gtcatccttg taatcgccct tatcgtcgtc atccttgtaa tctccagaac 4141 cacctccacc ggaacctcct ccgctcccta actgcttgtc accttctttc ttcagtcgac 4201 tggaaagaca aaacaggaaa gcgtatttag tctcaggcct aggcaggcag tcacgcaggc 4261 tggaggcaac cccaaagaca acagccaggc aatccaaccc caagcccgca gcctcctgtg 4321 tagctgacct accctcccaa cacctccctt cctctgtttc tcccatcacc atggttccac 4381 aatgtcccct ccagggccca gcaggcagag ggggaggagc ctggccctgg gcacacgggg 4441 aggttgggag cctgggctca gataacccct ccactccctc ccagcacccc acctcacctg 4501 taataatctt cctgacttgg taggtggcca ccatgcatgt gggggtgtcc atgcagccac 4561 tgctgctcca tggccagtct ctgcagctcg gccgactggg catgcatggc atgcagctgg 4621 tgggctgctg acatgggggg tgggatggcc ccaggcaggt cacgggggta gggggtgcct 4681 gccgaacaca aaacaacctg cgctacagaa atagccagag gactggccaa gaccagggcg 4741 gcacaggcac agcaaacggc aaagagcaca ggagaggtgc cagttcggac acaggagcca 4801 gcccctccca gactctgcag acagccagag ggcagagcct ctggggaggc aggaggtgcc 4861 cttgggtgcc tctagctggg ctgcaggtcg gggagaggag tgttagtaac cgcgacgctg 4921 gtgggggtcg gcttgttaag aggggcgctg ctaacgctgc aagagtgggt tgtcagcgtg 4981 gggccggtgc tactggaatc gataccggca tgattgacag cctgggcgag gatgtcacct 5041 gatggtgata agaatacacg ggagacttag tacggtttca caggcgtgac acgtttattg 5101 agtaggatta cagagtataa catagagtat aatatagagt atacaatagt gacgtgggat 5161 cctcagttag cctcccccat ctcccgggca aacgtgcgcg ccaggtcgca gatcgtcggt 5221 atggagccgg gggtggtgac gtgggtctgg accatcccgg aggtaagttg cagcagggcg 5281 tcccggcagc cggcgggcga ttggtcgtaa tccaggataa agacgtgcat gggacggagg 5341 cgtttggcca agacgtccaa ggcccaggca aacacgttgt acaggtcgcc gttgggggcc 5401 agcaactcgg gggcccgaaa cagggtaaat aacgtgtccc cgatatgggg tcgtgggccc 5461 gcgttgctct ggggctcggc accctggggc ggcacggccg tccccgaaag ctgtccccaa 5521 tcctcccgcc acgacccgcc gccctgcaga taccgcaccg tattggcaag cagcccgtaa 5581 acgcggcgaa tcgcggccag catagccagg tcaagccgct cgccggggcg ctggcgtttg 5641 gccaggcggt cgatgtgtct gtcctccgga agggccccca acacgatgtt tgtgccgggc 5701 aaggtcggcg ggatgagggc cacgaacgcc agcacggcct ggggggtcat gctgcccata 5761 aggtatcgcg cggccgggta gcacaggagg gcggcgatgg gatggcggtc gaagatgagg 5821 gtgagggccg ggggcggggc atgtgagctc ccagcctccc ccccgatatg aggagccaga 5881 acggcgtcgg tcacggcata aggcatgccc attgttatct gggcgcttgt cattaccacc 5941 gccgcgtccc cggccgatat ctcaccctgg tcgaggcggt gttgtgtggt gtagatgttc 6001 gcgattgtct cggaagcccc cagcacctgc cagtaagtca tcggctcggg tacgtagacg 6061 atatcgtcgc gcgaacccag ggccaccagc agttgcgtgg tggtggtttt ccccatcccg 6121 tgaggaccgt ctatataaac ccgcagtagc gtgggcattt tctgctccag gcggacttcc 6181 gtggcttctt gctgccggcg agggcgcaac gccgtacgtc ggttgctatg gccgcgagaa 6241 cgcgcagcct ggtcgaacgc agacgcgtgt tgatggcagg ggtacgaagc catggtcggg 6301 atccccaggc tggatcggtc ccggtgtctc ctatggaggt caaaacagcg tggatggcgt 6361 ctccaggcga tctgacggtt cactaaacga gctctgctta tatagacctc ccaccgtaca 6421 cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt 6481 tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact tgaaatcccc 6541 gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa 6601 tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact 6661 gggcataatg ccagggaatt gacctgcagg catgcaagct aattcgagct cggtacccgg 6721 ggatcctcta gagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt 6781 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa 6841 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact 6901 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 6961 ggggaga //