LOCUS Exported 6795 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Donor vector for 3' FLAG tag of human ZFP1_iso1. ACCESSION . VERSION . KEYWORDS pFETCh_ZFP1_iso1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6795) 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 6795) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6795 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 570..593 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 597..620 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 624..647 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 663..719 /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 732..1526 /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(786..805) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 1396..1415 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(2297..2314) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2297..2313) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2306..2328) /label=M13/pUC Forward /note="In lacZ gene" CDS complement(2532..3347) /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 3255..3274 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 3339..3358 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 3727..4315 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4216..4235 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4469..4486 /label=L4440 /note="L4440 vector, forward primer" protein_bind 4786..4807 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4822..4852 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4860..4876 /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 4865..4887 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 4884..4900 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4884..4900 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 5077..5279 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5230..5250 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 5276..5300 /label=LNCX /note="Human CMV promoter, forward primer" CDS 5376..6506 /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 atcagagaat tcacactggg gagaaacctt tcgagtgtcc ggaatgtgga aaagctttca 61 cccaccagtc aaacctcatt gtacaccaga gagcacatat ggagaagaag ccctatgagt 121 gcagtgaatg tggaaagaca tttgcccaaa agtttgaact caccacacac cagagaattc 181 atacaggaga gcgaccctat gagtgtaacg aatgtgcaaa aaccttcttt aagaagtcaa 241 accttatcat acatcagaag attcacacgg gggagaaacg ctatgagtgc agtgaatgtg 301 gaaaatcctt tatccagaac tcacagctca tcatacacat gagaactcat acaggagaga 361 aaccctatga atgtactgag tgcggcaaaa ctttcagcca gaggtcaact cttagattac 421 acttgcgaat ccacacagga gagaaaccat atgagtgttc cgaatgtggg aaggccttta 481 gcaggaagtc ccgactcagt gtccatcaga gagttcacat cggggagaaa cccgggagcg 541 gaggaggttc cggtggaggt ggttctggag attacaagga tgacgacgat aagggcgatt 601 acaaggatga cgacgataag ggagattaca aggatgacga cgataaggtt tcaggaagcg 661 gagctactaa cttcagcctg ctgaagcagg ctggagacgt ggaggagaac cctggacctg 721 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 781 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 841 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 901 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 961 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 1021 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 1081 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 1141 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 1201 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc 1261 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 1321 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggccgacc 1381 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 1441 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 1501 atcgccttct tgacgagttc ttctgattcg aacatcctgt ggaaaactcc tgccagaact 1561 cttcaagcgg gtgaaaaacc tcatgacagt attgagggaa catgggaatt catactgaga 1621 tgcaatctct caactcaaaa atgtattaaa aataggatcc catgagaaca ttatactgga 1681 agttacatgt gatacccagc taaagaatac atatcagaat atatccagtt gtaaatagcc 1741 atacccagtt gtactacaat gagcttttta aaatcttgaa tgaattgagt attctagaca 1801 gcagcataag aaatatacaa acagtatcct atgaatttag tggttttatg tggatatgca 1861 acaaaacaca agtggtatgc tttagatctg cacatgtgaa tttagcataa tcactgggaa 1921 tttgaaattc ttgtcctctg tgataattag gacataacaa gattttccat actaaacatc 1981 acatgtgttt ttcagtatct ctgcaatcca gtatgcattc caaatgaaat gtgtaacatt 2041 taaagtagca tggaacctac gtctttccct actgcttgct ggcatatatc agttggtatg 2101 aacattgttc ttgagctgcc tcttaggaaa cagaagacag tgttgaagtt ttccctgctt 2161 ctggtttgct gaagattttc tagaagcact atttacacaa atatgcaaat gtgaaataac 2221 cgatctataa cgtgaaggag gcagcttgga aagtcctctc cactgactgt agcctccaat 2281 tcactggccg tcgttcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg 2341 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag 2401 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgcgatttat 2461 tcaacaaagc cgccgtcccg tcaagtcagc gtaatgctct gccagtgtta caaccaatta 2521 accaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca 2581 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg 2641 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca 2701 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca 2761 tgagtgacga ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt 2821 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc 2881 attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa 2941 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct 3001 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt 3061 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc 3121 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca 3181 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct 3241 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa 3301 tttaatcgcg gcttcgagca agacgtttcc cgttgaatat ggctcataac accccttgta 3361 ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca 3421 atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct 3481 gagttgaagg atcagatcac gcatcttccc gacaacgcag accgttccgt ggcaaagcaa 3541 aagttcaaaa tcaccaactg gtccacctac aacaaagctc tcatcaaccg tggctccctc 3601 actttctggc tggatgatgg ggcgattcag gcctggtatg agtcagcaac accttcttca 3661 cgaggcagac ctctcgacgg agttccactg agcgtcagac cccgtagaaa agatcaaagg 3721 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc 3781 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac 3841 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca 3901 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt 3961 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc 4021 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg 4081 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc 4141 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac 4201 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct 4261 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc 4321 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt 4381 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac 4441 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagaagc 4501 tcgcacattc agcagcgttt ttcagcgcgt tttcgatcaa cgtttcaatg ttggtatcaa 4561 caccaggttt aactttgaac ttatcggcac tgacggttac tgattttgaa cttttgcttt 4621 gccacggaac ggtctgcgtt gtcgggaaga tgcgtgatct gatccttcaa ctcagcaaaa 4681 gttcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 4741 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct 4801 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat 4861 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc 4921 atgcctgcag gtcgactcta gaggatcccc gggtaccgag ctcgaattag cttgcatgcc 4981 tgcaggtcaa ttccctggca ttatgcccag tacatgacct tatgggactt tcctacttgg 5041 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc 5101 aatgggcgtg gatagcggtt tgactcacgg ggatttcaag tctccacccc attgacgtca 5161 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg 5221 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc 5281 gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagga 5341 gacaccggga ccgatccagc ctggggatcc cgaccatggc ttcgtacccc tgccatcaac 5401 acgcgtctgc gttcgaccag gctgcgcgtt ctcgcggcca tagcaaccga cgtacggcgt 5461 tgcgccctcg ccggcagcaa gaagccacgg aagtccgcct ggagcagaaa atgcccacgc 5521 tactgcgggt ttatatagac ggtcctcacg ggatggggaa aaccaccacc acgcaactgc 5581 tggtggccct gggttcgcgc gacgatatcg tctacgtacc cgagccgatg acttactggc 5641 aggtgctggg ggcttccgag acaatcgcga acatctacac cacacaacac cgcctcgacc 5701 agggtgagat atcggccggg gacgcggcgg tggtaatgac aagcgcccag ataacaatgg 5761 gcatgcctta tgccgtgacc gacgccgttc tggctcctca tatcgggggg gaggctggga 5821 gctcacatgc cccgcccccg gccctcaccc tcatcttcga ccgccatccc atcgccgccc 5881 tcctgtgcta cccggccgcg cgatacctta tgggcagcat gaccccccag gccgtgctgg 5941 cgttcgtggc cctcatcccg ccgaccttgc ccggcacaaa catcgtgttg ggggcccttc 6001 cggaggacag acacatcgac cgcctggcca aacgccagcg ccccggcgag cggcttgacc 6061 tggctatgct ggccgcgatt cgccgcgttt acgggctgct tgccaatacg gtgcggtatc 6121 tgcagggcgg cgggtcgtgg cgggaggatt ggggacagct ttcggggacg gccgtgccgc 6181 cccagggtgc cgagccccag agcaacgcgg gcccacgacc ccatatcggg gacacgttat 6241 ttaccctgtt tcgggccccc gagttgctgg cccccaacgg cgacctgtac aacgtgtttg 6301 cctgggcctt ggacgtcttg gccaaacgcc tccgtcccat gcacgtcttt atcctggatt 6361 acgaccaatc gcccgccggc tgccgggacg ccctgctgca acttacctcc gggatggtcc 6421 agacccacgt caccaccccc ggctccatac cgacgatctg cgacctggcg cgcacgtttg 6481 cccgggagat gggggaggct aactgaggat cccacgtcac tattgtatac tctatattat 6541 actctatgtt atactctgta atcctactca ataaacgtgt cacgcctgtg aaaccgtact 6601 aagtctcccg tgtattctta tcaccatcag gtgacatcct cgcccaggct gtcaatcatg 6661 ccggtatcga ttccagtagc accggcccca cgctgacaac ccactcttgc agcgttagca 6721 gcgcccctct taacaagccg acccccacca gcgtcgcggt tactaacact cctctccccg 6781 acctgcagcc cagct //