LOCUS Exported 6752 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Donor vector for 3' FLAG tag of human GATAD2A. ACCESSION . VERSION . KEYWORDS pFETCh_GATAD2A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6752) 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 6752) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6752 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 743..766 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 770..793 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 797..820 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 836..892 /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 905..1699 /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(959..978) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 1569..1588 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(2254..2271) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2254..2270) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2263..2285) /label=M13/pUC Forward /note="In lacZ gene" CDS complement(2489..3304) /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 3212..3231 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 3296..3315 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 3684..4272 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4173..4192 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4426..4443 /label=L4440 /note="L4440 vector, forward primer" protein_bind 4743..4764 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4779..4809 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4817..4833 /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 4822..4844 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 4841..4857 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4841..4857 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 5034..5236 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5187..5207 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 5233..5257 /label=LNCX /note="Human CMV promoter, forward primer" CDS 5333..6463 /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 tcagcctctg gagtaactgg gactacaggg atgtgccacc atgtccagct gatatttttt 61 ttagagatgg ggcctcacta tgttgcccag gctggtctct atctcctggc ctcaagcagt 121 cttcccaccc cccagcctct caaagtgctg ggattcaggc tgagcctccg tgcccagcta 181 agatagtttt tcattgctgc atagtattcc atcctgcatt attgtcctgt tgcacacatt 241 tgtttatggt tttccactgt tctaagcagc accgtggtca gcacttcatt tctgtagaac 301 gtaggtagct gctcaccctc tgatgattgc cccacacttg ggattacctt ggcagagcct 361 ggcaatgtca aggccattga gttgctgcct aaatactctt cagctctggg ggggcctggg 421 atttgaaacg ttgattgagg agtagtgtgg gtgggtttgc aagggctcct gggtccttag 481 tctgtattgg ggctggggtc taggcagcta tggctgggct cctcccagga gccctcctga 541 cgagggcctt ctcagctggt cgctctgttc tgttgcaggc gggacccttg cgtttgtcag 601 cccaagcctg gcggtgcaca agagctcctc ggccgtggac cgccagcgag agtacctcct 661 ggacatgatc ccaccccgct ccatccccca gtcagccacg tggaaaggga gcggaggagg 721 ttccggtgga ggtggttctg gagattacaa ggatgacgac gataagggcg attacaagga 781 tgacgacgat aagggagatt acaaggatga cgacgataag gtttcaggaa gcggagctac 841 taacttcagc ctgctgaagc aggctggaga cgtggaggag aaccctggac ctggatcgtt 901 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct 961 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct 1021 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga 1081 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc 1141 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg 1201 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc 1261 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca 1321 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga 1381 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc 1441 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga 1501 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggccg accgctatca 1561 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg 1621 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct 1681 tcttgacgag ttcttctgat tcgaacatcg gaagacgggc tccctcctcc cccacctggc 1741 ccctggtcta gaaggaccca ctgcaccacc ctccgctggc tcgggaagac accgtgcccg 1801 ccccaagagc aagcaccggc catgctgcag aggcaagacc tcaattcttg gctgcaaagt 1861 ttcatcaggg ctagggggct ggtgccgcct cataggcaga cgaggatcat cgctggggga 1921 cctttcccgt gggctttctt cctttctctc tttgccttta gtttgcccga caccagcaga 1981 aaagtggacc ttgggggctg gttctgctcc tggccccctt gttcagcccc tgccggcaca 2041 cgggcggctc accctggaca ctgtgatgcg catgggcaag gccagcgccc ggggcttctg 2101 aaccgagcgg ggtgtttcat ttttttgctt ttccctgtct taggctccca gtctttgact 2161 gccttcccat ggcgatctat aagttgaaag attttttttt tcttggaaag tcctctccac 2221 tgactgtagc ctccaattca ctggccgtcg ttcactggcc gtcgttttac aacgtcgtga 2281 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag 2341 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa 2401 tggcgaatgc gatttattca acaaagccgc cgtcccgtca agtcagcgta atgctctgcc 2461 agtgttacaa ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact 2521 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg 2581 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga 2641 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat 2701 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca 2761 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat 2821 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt 2881 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat 2941 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg 3001 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg 3061 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg 3121 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc 3181 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat 3241 cagcatccat gttggaattt aatcgcggct tcgagcaaga cgtttcccgt tgaatatggc 3301 tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata 3361 tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa 3421 taaatcgaac ttttgctgag ttgaaggatc agatcacgca tcttcccgac aacgcagacc 3481 gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc cacctacaac aaagctctca 3541 tcaaccgtgg ctccctcact ttctggctgg atgatggggc gattcaggcc tggtatgagt 3601 cagcaacacc ttcttcacga ggcagacctc tcgacggagt tccactgagc gtcagacccc 3661 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg 3721 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact 3781 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg 3841 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg 3901 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac 3961 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca 4021 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga 4081 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc 4141 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct 4201 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg 4261 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct 4321 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc 4381 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc 4441 gaggaagcgg aagaagctcg cacattcagc agcgtttttc agcgcgtttt cgatcaacgt 4501 ttcaatgttg gtatcaacac caggtttaac tttgaactta tcggcactga cggttactga 4561 ttttgaactt ttgctttgcc acggaacggt ctgcgttgtc gggaagatgc gtgatctgat 4621 ccttcaactc agcaaaagtt cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt 4681 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca 4741 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct 4801 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat 4861 gattacgcca agcttgcatg cctgcaggtc gactctagag gatccccggg taccgagctc 4921 gaattagctt gcatgcctgc aggtcaattc cctggcatta tgcccagtac atgaccttat 4981 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc 5041 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttcaagtct 5101 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa 5161 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt 5221 ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg 5281 ttttgacctc cataggagac accgggaccg atccagcctg gggatcccga ccatggcttc 5341 gtacccctgc catcaacacg cgtctgcgtt cgaccaggct gcgcgttctc gcggccatag 5401 caaccgacgt acggcgttgc gccctcgccg gcagcaagaa gccacggaag tccgcctgga 5461 gcagaaaatg cccacgctac tgcgggttta tatagacggt cctcacggga tggggaaaac 5521 caccaccacg caactgctgg tggccctggg ttcgcgcgac gatatcgtct acgtacccga 5581 gccgatgact tactggcagg tgctgggggc ttccgagaca atcgcgaaca tctacaccac 5641 acaacaccgc ctcgaccagg gtgagatatc ggccggggac gcggcggtgg taatgacaag 5701 cgcccagata acaatgggca tgccttatgc cgtgaccgac gccgttctgg ctcctcatat 5761 cgggggggag gctgggagct cacatgcccc gcccccggcc ctcaccctca tcttcgaccg 5821 ccatcccatc gccgccctcc tgtgctaccc ggccgcgcga taccttatgg gcagcatgac 5881 cccccaggcc gtgctggcgt tcgtggccct catcccgccg accttgcccg gcacaaacat 5941 cgtgttgggg gcccttccgg aggacagaca catcgaccgc ctggccaaac gccagcgccc 6001 cggcgagcgg cttgacctgg ctatgctggc cgcgattcgc cgcgtttacg ggctgcttgc 6061 caatacggtg cggtatctgc agggcggcgg gtcgtggcgg gaggattggg gacagctttc 6121 ggggacggcc gtgccgcccc agggtgccga gccccagagc aacgcgggcc cacgacccca 6181 tatcggggac acgttattta ccctgtttcg ggcccccgag ttgctggccc ccaacggcga 6241 cctgtacaac gtgtttgcct gggccttgga cgtcttggcc aaacgcctcc gtcccatgca 6301 cgtctttatc ctggattacg accaatcgcc cgccggctgc cgggacgccc tgctgcaact 6361 tacctccggg atggtccaga cccacgtcac cacccccggc tccataccga cgatctgcga 6421 cctggcgcgc acgtttgccc gggagatggg ggaggctaac tgaggatccc acgtcactat 6481 tgtatactct atattatact ctatgttata ctctgtaatc ctactcaata aacgtgtcac 6541 gcctgtgaaa ccgtactaag tctcccgtgt attcttatca ccatcaggtg acatcctcgc 6601 ccaggctgtc aatcatgccg gtatcgattc cagtagcacc ggccccacgc tgacaaccca 6661 ctcttgcagc gttagcagcg cccctcttaa caagccgacc cccaccagcg tcgcggttac 6721 taacactcct ctccccgacc tgcagcccag ct //