LOCUS Exported 6970 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Donor vector for 3' FLAG tag of human SAP130. ACCESSION . VERSION . KEYWORDS pFETCh_SAP130 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6970) 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 6970) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6970 /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(2472..2489) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2472..2488) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2481..2503) /label=M13/pUC Forward /note="In lacZ gene" CDS complement(2707..3522) /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 3430..3449 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 3514..3533 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 3902..4490 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4391..4410 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4644..4661 /label=L4440 /note="L4440 vector, forward primer" protein_bind 4961..4982 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4997..5027 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5035..5051 /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 5040..5062 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5059..5075 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5059..5075 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 5252..5454 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5405..5425 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 5451..5475 /label=LNCX /note="Human CMV promoter, forward primer" CDS 5551..6681 /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 gcgcccagcc cctttactgt ttttaaacct tagtatctta gtccttagga gaaaggacgt 61 tgtcacccaa acaactctta gacagtttac aaaggtgatt gaaggtgggt tcatttgcct 121 ggggttgaca tagaaaaata tttccgagct tatgataccc tcccttttgt gtgttgcaga 181 cgaatctaga acatgatgtc tatgaaagac ttactaacct gcaggaaggg attatcccaa 241 agaaaaaagc agcaacagat gatgatctcc accgaataaa cgaactgata caggtttgag 301 tgcgcccctt cccttctgat catctacttt atgaagttgc tttctgtaaa gccctacttc 361 tcccgtttcc ccccgtctcc ctcatatctt cagtaaaaac atacgtgctt cagcctgcac 421 tcctgagaaa caaaaagaca ttttctctgt tgtggaagcc tcaacatggc ctctgcccaa 481 aggcaagcag ttttttaaaa gctgtactat cctcatggtc actgattgat gatgcaatgt 541 cttctgtttc agggaaatat gcagaggtgt aaacttgtga tggatcaaat cagtgaagcc 601 agagactcca tgcttaaggt tttagatcat aaagaccgtg tcctgaagct gcttaacaag 661 aacgggactg tcaaaaaagt gtccaaattg aagcgaaagg aaaaagtcgg 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 catcaggaga ttggaagcaa atttatgaag 1741 aatgatggtg ggggtggggg gagggttttg gttttttcca aagtggaaca ttgaaataaa 1801 ggaagtgttc cttagttccc gtgtgaaagc agaggaaccc atgacatcca agggcgtgaa 1861 aggatcagag ctgactggac atagtgagct gccttcttgc gttcgggtgc acccctgtta 1921 aacctgatct gtgtcataag tgactccgga tgcatcagtg tccaccagtt ggaagcaatg 1981 acaaggatgg ctggctggtg tttttcagcc ttccggttta tagactgtat ttatctagtg 2041 gattcctgca ggccccatac tgagcctgga ctgaaagtat ccactcggac catctgttat 2101 ctctctacac tgaaaataaa acctcttcca cccaccccat tcggttcttc tgcctgacct 2161 tcaaatgccc atgttggcct tttacagcag tgccacggca ccaagcgagc tgccacatct 2221 cacactctaa agggtttgaa ctattagttc ttgtcatttt ttaaaaaaaa ccattcccaa 2281 gtgaaattgt tatatcgtct gtcttgcgtg tgtcagaact gggtttttgt ggaggttcag 2341 agcaggcaac accataagtt gctctcagat ccttgttctg aagtacattc ttggttatct 2401 gtacttctgt agctggtgtc ttggaaagtc ctctccactg actgtagcct ccaattcact 2461 ggccgtcgtt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc 2521 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc 2581 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatgcga tttattcaac 2641 aaagccgccg tcccgtcaag tcagcgtaat gctctgccag tgttacaacc aattaaccaa 2701 ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt 2761 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca 2821 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat 2881 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt 2941 gacgactgaa tccggtgaga atggcaaaag cttatgcatt tctttccaga cttgttcaac 3001 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg 3061 tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg 3121 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc 3181 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca 3241 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag 3301 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt 3361 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg 3421 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa 3481 tcgcggcttc gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact 3541 gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta 3601 acatcagaga ttttgagaca caacgtggct ttgttgaata aatcgaactt ttgctgagtt 3661 gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt tccgtggcaa agcaaaagtt 3721 caaaatcacc aactggtcca cctacaacaa agctctcatc aaccgtggct ccctcacttt 3781 ctggctggat gatggggcga ttcaggcctg gtatgagtca gcaacacctt cttcacgagg 3841 cagacctctc gacggagttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt 3901 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 3961 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 4021 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact 4081 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 4141 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 4201 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 4261 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 4321 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 4381 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 4441 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 4501 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg 4561 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc 4621 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gaagctcgca 4681 cattcagcag cgtttttcag cgcgttttcg atcaacgttt caatgttggt atcaacacca 4741 ggtttaactt tgaacttatc ggcactgacg gttactgatt ttgaactttt gctttgccac 4801 ggaacggtct gcgttgtcgg gaagatgcgt gatctgatcc ttcaactcag caaaagttcc 4861 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca 4921 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc 4981 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga 5041 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cttgcatgcc 5101 tgcaggtcga ctctagagga tccccgggta ccgagctcga attagcttgc atgcctgcag 5161 gtcaattccc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta 5221 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg 5281 gcgtggatag cggtttgact cacggggatt tcaagtctcc accccattga cgtcaatggg 5341 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca 5401 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta 5461 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca taggagacac 5521 cgggaccgat ccagcctggg gatcccgacc atggcttcgt acccctgcca tcaacacgcg 5581 tctgcgttcg accaggctgc gcgttctcgc ggccatagca accgacgtac ggcgttgcgc 5641 cctcgccggc agcaagaagc cacggaagtc cgcctggagc agaaaatgcc cacgctactg 5701 cgggtttata tagacggtcc tcacgggatg gggaaaacca ccaccacgca actgctggtg 5761 gccctgggtt cgcgcgacga tatcgtctac gtacccgagc cgatgactta ctggcaggtg 5821 ctgggggctt ccgagacaat cgcgaacatc tacaccacac aacaccgcct cgaccagggt 5881 gagatatcgg ccggggacgc ggcggtggta atgacaagcg cccagataac aatgggcatg 5941 ccttatgccg tgaccgacgc cgttctggct cctcatatcg ggggggaggc tgggagctca 6001 catgccccgc ccccggccct caccctcatc ttcgaccgcc atcccatcgc cgccctcctg 6061 tgctacccgg ccgcgcgata ccttatgggc agcatgaccc cccaggccgt gctggcgttc 6121 gtggccctca tcccgccgac cttgcccggc acaaacatcg tgttgggggc ccttccggag 6181 gacagacaca tcgaccgcct ggccaaacgc cagcgccccg gcgagcggct tgacctggct 6241 atgctggccg cgattcgccg cgtttacggg ctgcttgcca atacggtgcg gtatctgcag 6301 ggcggcgggt cgtggcggga ggattgggga cagctttcgg ggacggccgt gccgccccag 6361 ggtgccgagc cccagagcaa cgcgggccca cgaccccata tcggggacac gttatttacc 6421 ctgtttcggg cccccgagtt gctggccccc aacggcgacc tgtacaacgt gtttgcctgg 6481 gccttggacg tcttggccaa acgcctccgt cccatgcacg tctttatcct ggattacgac 6541 caatcgcccg ccggctgccg ggacgccctg ctgcaactta cctccgggat ggtccagacc 6601 cacgtcacca cccccggctc cataccgacg atctgcgacc tggcgcgcac gtttgcccgg 6661 gagatggggg aggctaactg aggatcccac gtcactattg tatactctat attatactct 6721 atgttatact ctgtaatcct actcaataaa cgtgtcacgc ctgtgaaacc gtactaagtc 6781 tcccgtgtat tcttatcacc atcaggtgac atcctcgccc aggctgtcaa tcatgccggt 6841 atcgattcca gtagcaccgg ccccacgctg acaacccact cttgcagcgt tagcagcgcc 6901 cctcttaaca agccgacccc caccagcgtc gcggttacta acactcctct ccccgacctg 6961 cagcccagct //