LOCUS Exported 7182 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Donor vector for 3' FLAG tag of human ZNF219 . ACCESSION . VERSION . KEYWORDS pFETCh_ZNF219 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7182) TITLE Mendenhall CRISPR tagging plasmids REFERENCE 2 (bases 1 to 7182) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7182 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 865..888 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 892..915 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 919..942 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 958..1014 /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 1027..1821 /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(1081..1100) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 1691..1710 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(2684..2701) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2684..2700) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2693..2715) /label=M13/pUC Forward /note="In lacZ gene" CDS complement(2919..3734) /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 3642..3661 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 3726..3745 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 4114..4702 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4603..4622 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4856..4873 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5173..5194 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5209..5239 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5247..5263 /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 5252..5274 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5271..5287 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5271..5287 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 5464..5666 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5617..5637 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 5663..5687 /label=LNCX /note="Human CMV promoter, forward primer" CDS 5763..6893 /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 cagaaaggtg agggctaccg aaacactagg aggtgaagtt tgttccgaag ccttcagttt 61 ggctggatga gatcacttgg gaaacgctcc tttattaaat caaagtgggt gggagacgca 121 gtgccctttc ttggttctga gaatgaggtc tgtaaattgg gcgaaattcg ggtgggtgcg 181 tgcagtcatc gtgaccccgt gaccctatcg cacccactct ccccaggcga gcggccctac 241 aagtgtccgc actgcgacta cgcgggcacc cagtccggct cgctcaagta tcacctacag 301 cgccaccacc gggagcagag gagcggggcc ggccccgggc cacccccgga gccaccgcct 361 ccttcccagc ggggttcggc cccgcaatct ggagccaagc cgtctccgca gcctgcgacc 421 tgggtggagg gcgcctcaag tccccggcct ccttctagcg gtgctgggcc ggggtcccgt 481 cggaagcccg ccagccctgg gaggaccctg cgcaacgggc gaggcggtga ggccgaaccc 541 ctggacctgt ccttgcgggc agggccggga ggcgaggccg ggcctggggg tgccctccac 601 cgctgcctct tctgcccgtt cgccactgga gccccagagc tcatggcctt gcaccttcaa 661 gtgcaccaca gccgccgggc taggggccgc cggccacccc aggctgacgc gtccccgccc 721 tatgcccgag taccatcagg agagacccct cccagtcctt cgcaggaagg ggaggagggc 781 tccgggctgt ccagacccgg agaggcaggg ctgggggggc aagaacgggg gagcggagga 841 ggttccggtg gaggtggttc tggagattac aaggatgacg acgataaggg cgattacaag 901 gatgacgacg ataagggaga ttacaaggat gacgacgata aggtttcagg aagcggagct 961 actaacttca gcctgctgaa gcaggctgga gacgtggagg agaaccctgg acctggatcg 1021 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg 1081 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg 1141 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat 1201 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca 1261 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg 1321 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat 1381 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa 1441 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg 1501 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg 1561 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg 1621 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc cgaccgctat 1681 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac 1741 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc 1801 cttcttgacg agttcttctg attcgaacat cggggcgatt agcttagtga gcttaccccg 1861 ccggagcggg ggagcgctag aggtagttgg gtagagaagc cagcaggggg cagcgtggga 1921 cccatatccc agccccaggc ggaccagagg tggaaggcgc ggcgggcgta ggagggtggg 1981 ctggcggtac ccacccagcc caagggagtc acagtgcctt agaagtggca gaggtgaggg 2041 tcaaagaacg gggtccctgg gctggcagag aaggtttgtg tccctgttga ggagccactc 2101 tgccagtctt tgctggtcca taggcgtgag agtttatttt tgtacaaggg gtgagggtgg 2161 ggggcactgt acccttctag ccccatcagg ggccctgtag acgtcgctat ttttggtacg 2221 attcctgtca tccctattgc agagtcgtgt ccccaataaa caggtgtctt gaggcacagg 2281 gccctgtgtc tgtctgccta cctatgtcct gcttccctct ttgggtcctt ctgtcaggaa 2341 atagtcacct cttcagacac ttagttatca cccaaagagt ttgataagaa atgcagaata 2401 ttgggtttta cccccaggtt tcgatgcagt agctctgatg gggcccaaga attagcattt 2461 aacaaacagc caagcaagtt ctcacgcact gtggcctgag gacctcattt tgagaactgc 2521 tggactgtgc tggatcccag tttgtctgtg accagttcag cctcaaggag taggtgcagt 2581 gtgggagaag gagcttcaga gagtgtgtgg aacctctggg agccactgtg gcttggaaag 2641 tcctctccac tgactgtagc ctccaattca ctggccgtcg ttcactggcc gtcgttttac 2701 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc 2761 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc 2821 gcagcctgaa tggcgaatgc gatttattca acaaagccgc cgtcccgtca agtcagcgta 2881 atgctctgcc agtgttacaa ccaattaacc aattctgatt agaaaaactc atcgagcatc 2941 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt 3001 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat 3061 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa 3121 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa 3181 agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa 3241 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg 3301 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact 3361 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct 3421 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc 3481 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta 3541 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc 3601 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac 3661 ccatataaat cagcatccat gttggaattt aatcgcggct tcgagcaaga cgtttcccgt 3721 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt 3781 catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg 3841 ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc agatcacgca tcttcccgac 3901 aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc cacctacaac 3961 aaagctctca tcaaccgtgg ctccctcact ttctggctgg atgatggggc gattcaggcc 4021 tggtatgagt cagcaacacc ttcttcacga ggcagacctc tcgacggagt tccactgagc 4081 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 4141 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 4201 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 4261 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 4321 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 4381 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 4441 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 4501 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 4561 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 4621 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 4681 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt 4741 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg 4801 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga 4861 gtcagtgagc gaggaagcgg aagaagctcg cacattcagc agcgtttttc agcgcgtttt 4921 cgatcaacgt ttcaatgttg gtatcaacac caggtttaac tttgaactta tcggcactga 4981 cggttactga ttttgaactt ttgctttgcc acggaacggt ctgcgttgtc gggaagatgc 5041 gtgatctgat ccttcaactc agcaaaagtt cccaatacgc aaaccgcctc tccccgcgcg 5101 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga 5161 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat 5221 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag 5281 ctatgaccat gattacgcca agcttgcatg cctgcaggtc gactctagag gatccccggg 5341 taccgagctc gaattagctt gcatgcctgc aggtcaattc cctggcatta tgcccagtac 5401 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc 5461 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga 5521 tttcaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 5581 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac 5641 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc 5701 atccacgctg ttttgacctc cataggagac accgggaccg atccagcctg gggatcccga 5761 ccatggcttc gtacccctgc catcaacacg cgtctgcgtt cgaccaggct gcgcgttctc 5821 gcggccatag caaccgacgt acggcgttgc gccctcgccg gcagcaagaa gccacggaag 5881 tccgcctgga gcagaaaatg cccacgctac tgcgggttta tatagacggt cctcacggga 5941 tggggaaaac caccaccacg caactgctgg tggccctggg ttcgcgcgac gatatcgtct 6001 acgtacccga gccgatgact tactggcagg tgctgggggc ttccgagaca atcgcgaaca 6061 tctacaccac acaacaccgc ctcgaccagg gtgagatatc ggccggggac gcggcggtgg 6121 taatgacaag cgcccagata acaatgggca tgccttatgc cgtgaccgac gccgttctgg 6181 ctcctcatat cgggggggag gctgggagct cacatgcccc gcccccggcc ctcaccctca 6241 tcttcgaccg ccatcccatc gccgccctcc tgtgctaccc ggccgcgcga taccttatgg 6301 gcagcatgac cccccaggcc gtgctggcgt tcgtggccct catcccgccg accttgcccg 6361 gcacaaacat cgtgttgggg gcccttccgg aggacagaca catcgaccgc ctggccaaac 6421 gccagcgccc cggcgagcgg cttgacctgg ctatgctggc cgcgattcgc cgcgtttacg 6481 ggctgcttgc caatacggtg cggtatctgc agggcggcgg gtcgtggcgg gaggattggg 6541 gacagctttc ggggacggcc gtgccgcccc agggtgccga gccccagagc aacgcgggcc 6601 cacgacccca tatcggggac acgttattta ccctgtttcg ggcccccgag ttgctggccc 6661 ccaacggcga cctgtacaac gtgtttgcct gggccttgga cgtcttggcc aaacgcctcc 6721 gtcccatgca cgtctttatc ctggattacg accaatcgcc cgccggctgc cgggacgccc 6781 tgctgcaact tacctccggg atggtccaga cccacgtcac cacccccggc tccataccga 6841 cgatctgcga cctggcgcgc acgtttgccc gggagatggg ggaggctaac tgaggatccc 6901 acgtcactat tgtatactct atattatact ctatgttata ctctgtaatc ctactcaata 6961 aacgtgtcac gcctgtgaaa ccgtactaag tctcccgtgt attcttatca ccatcaggtg 7021 acatcctcgc ccaggctgtc aatcatgccg gtatcgattc cagtagcacc ggccccacgc 7081 tgacaaccca ctcttgcagc gttagcagcg cccctcttaa caagccgacc cccaccagcg 7141 tcgcggttac taacactcct ctccccgacc tgcagcccag ct //