LOCUS Exported 6892 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Donor vector for 3' FLAG tag of human ARID5B. ACCESSION . VERSION . KEYWORDS pFETCh_ARID5B SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6892) 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 6892) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6892 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 745..768 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 772..795 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 799..822 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 838..894 /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 907..1701 /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(961..980) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 1571..1590 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(2394..2411) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2394..2410) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2403..2425) /label=M13/pUC Forward /note="In lacZ gene" CDS complement(2629..3444) /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 3352..3371 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 3436..3455 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 3824..4412 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4313..4332 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4566..4583 /label=L4440 /note="L4440 vector, forward primer" protein_bind 4883..4904 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4919..4949 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4957..4973 /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 4962..4984 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 4981..4997 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4981..4997 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 5174..5376 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5327..5347 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 5373..5397 /label=LNCX /note="Human CMV promoter, forward primer" CDS 5473..6603 /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 aaagcctgtc gggtatcacc catgaccatg tcaggcccta aaaaataccc tgaatcgctt 61 tcaagatcag gaaaacctca ccatgtgaga ctggagaatt tcaggaagat ggaaggcatg 121 gtccacccaa tcctgcaccg gaaaatgagc ccgcagaaca ttggggcggc gcggccgatc 181 aagcgcagcc tggaggattt ggaccttgtg attgcaggga aaaaggcccg ggcagtgtct 241 cccttagacc catccaagga ggtctctggg aaggagaagg cctctgagca ggagagtgaa 301 ggcagcaaag cagcgcacgg tgggcattcc gggggcggat cagaaggcca caagcttccc 361 ctctcctccc ctatcttccc aggtctgtat tccgggagcc tgtgtaactc gggcctcaac 421 tccaggctcc cggctgggta ttctcattct ctgcagtact tgaaaaacca gactgtgctt 481 tctccactca tgcagcccct ggctttccac tcgcttgtga tgcaaagagg aatttttaca 541 tcaccgacaa attctcagca gctgtacaga cacttggctg cggctacacc tgtaggaagt 601 tcatatgggg accttttgca taacagcatt taccctttag ctgctataaa tcctcaagct 661 gcctttccat cttcccagct gtcatccgtg caccccagta ctaagctcgg gagcggagga 721 ggttccggtg gaggtggttc tggagattac aaggatgacg acgataaggg cgattacaag 781 gatgacgacg ataagggaga ttacaaggat gacgacgata aggtttcagg aagcggagct 841 actaacttca gcctgctgaa gcaggctgga gacgtggagg agaaccctgg acctggatcg 901 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg 961 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg 1021 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat 1081 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca 1141 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg 1201 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat 1261 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa 1321 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg 1381 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg 1441 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg 1501 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc cgaccgctat 1561 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac 1621 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc 1681 cttcttgacg agttcttctg attcgaacat cgctcagctc tgcccagcag tccaaagcgg 1741 catggccaac agagcttcac tccttaccca ggagtgctgg cttatagagt tagaagtcag 1801 tatttcttct aatctgaggc tatgatcagt cccagctgta ggggcccaga ggggaggtga 1861 acatgcctga tttttgtggg acaactctag cccacaaact gactggctgg tgagtcttga 1921 ctcccttcca acacagatgc ccaggcacct ccagatcatt cacttcgcac gtgggccttg 1981 tgaagggatt tgtgaatatc caggaagaac ttagaggacc ccatctgagt tcggatggtc 2041 aggaaacaat ctgggcaaaa aagaggcagg catttcaaag gaaggggcaa ggaagactgg 2101 caaacagatg gcaagggatg cccctctttt tcataaaact ctccaaggtt caatcaatgc 2161 aatgtatagt gaaacttcaa tagatctttc attttgacac tattaaacaa tccagagaag 2221 taaacactgt taaattgact gtatatattt gcttcttaaa actacctgta tcactgtttg 2281 ctcacctaat ttatatacag gtagttccat tttctcccag ttccttctcg tctttttttt 2341 tcttggaaag tcctctccac tgactgtagc ctccaattca ctggccgtcg ttcactggcc 2401 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca 2461 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc 2521 caacagttgc gcagcctgaa tggcgaatgc gatttattca acaaagccgc cgtcccgtca 2581 agtcagcgta atgctctgcc agtgttacaa ccaattaacc aattctgatt agaaaaactc 2641 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg 2701 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag 2761 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc 2821 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga 2881 gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc 2941 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag 3001 acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg 3061 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac 3121 ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg 3181 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat 3241 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc 3301 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc 3361 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggct tcgagcaaga 3421 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag 3481 ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga 3541 cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc agatcacgca 3601 tcttcccgac aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc 3661 cacctacaac aaagctctca tcaaccgtgg ctccctcact ttctggctgg atgatggggc 3721 gattcaggcc tggtatgagt cagcaacacc ttcttcacga ggcagacctc tcgacggagt 3781 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc 3841 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc 3901 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac 3961 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac 4021 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt 4081 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct 4141 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat 4201 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt 4261 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg 4321 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt 4381 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt 4441 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg 4501 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg 4561 agcgcagcga gtcagtgagc gaggaagcgg aagaagctcg cacattcagc agcgtttttc 4621 agcgcgtttt cgatcaacgt ttcaatgttg gtatcaacac caggtttaac tttgaactta 4681 tcggcactga cggttactga ttttgaactt ttgctttgcc acggaacggt ctgcgttgtc 4741 gggaagatgc gtgatctgat ccttcaactc agcaaaagtt cccaatacgc aaaccgcctc 4801 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag 4861 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt 4921 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca 4981 caggaaacag ctatgaccat gattacgcca agcttgcatg cctgcaggtc gactctagag 5041 gatccccggg taccgagctc gaattagctt gcatgcctgc aggtcaattc cctggcatta 5101 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat 5161 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga 5221 ctcacgggga tttcaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa 5281 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt 5341 aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc 5401 tggagacgcc atccacgctg ttttgacctc cataggagac accgggaccg atccagcctg 5461 gggatcccga ccatggcttc gtacccctgc catcaacacg cgtctgcgtt cgaccaggct 5521 gcgcgttctc gcggccatag caaccgacgt acggcgttgc gccctcgccg gcagcaagaa 5581 gccacggaag tccgcctgga gcagaaaatg cccacgctac tgcgggttta tatagacggt 5641 cctcacggga tggggaaaac caccaccacg caactgctgg tggccctggg ttcgcgcgac 5701 gatatcgtct acgtacccga gccgatgact tactggcagg tgctgggggc ttccgagaca 5761 atcgcgaaca tctacaccac acaacaccgc ctcgaccagg gtgagatatc ggccggggac 5821 gcggcggtgg taatgacaag cgcccagata acaatgggca tgccttatgc cgtgaccgac 5881 gccgttctgg ctcctcatat cgggggggag gctgggagct cacatgcccc gcccccggcc 5941 ctcaccctca tcttcgaccg ccatcccatc gccgccctcc tgtgctaccc ggccgcgcga 6001 taccttatgg gcagcatgac cccccaggcc gtgctggcgt tcgtggccct catcccgccg 6061 accttgcccg gcacaaacat cgtgttgggg gcccttccgg aggacagaca catcgaccgc 6121 ctggccaaac gccagcgccc cggcgagcgg cttgacctgg ctatgctggc cgcgattcgc 6181 cgcgtttacg ggctgcttgc caatacggtg cggtatctgc agggcggcgg gtcgtggcgg 6241 gaggattggg gacagctttc ggggacggcc gtgccgcccc agggtgccga gccccagagc 6301 aacgcgggcc cacgacccca tatcggggac acgttattta ccctgtttcg ggcccccgag 6361 ttgctggccc ccaacggcga cctgtacaac gtgtttgcct gggccttgga cgtcttggcc 6421 aaacgcctcc gtcccatgca cgtctttatc ctggattacg accaatcgcc cgccggctgc 6481 cgggacgccc tgctgcaact tacctccggg atggtccaga cccacgtcac cacccccggc 6541 tccataccga cgatctgcga cctggcgcgc acgtttgccc gggagatggg ggaggctaac 6601 tgaggatccc acgtcactat tgtatactct atattatact ctatgttata ctctgtaatc 6661 ctactcaata aacgtgtcac gcctgtgaaa ccgtactaag tctcccgtgt attcttatca 6721 ccatcaggtg acatcctcgc ccaggctgtc aatcatgccg gtatcgattc cagtagcacc 6781 ggccccacgc tgacaaccca ctcttgcagc gttagcagcg cccctcttaa caagccgacc 6841 cccaccagcg tcgcggttac taacactcct ctccccgacc tgcagcccag ct //