LOCUS       sequence-260781-        6733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human ZNF326_iso1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6733)
  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 6733)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genome Res.
            2015 Sep 9."
COMMENT     Sequence Label: pFETCh_ZNF326_iso1
FEATURES             Location/Qualifiers
     source          1..6733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             97..1227
                     /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"
     CDS             2074..2097
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2101..2124
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2128..2151
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2167..2223
                     /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             2236..3030
                     /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(2290..2309)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2900..2919
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3751..3767)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3760..3782)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(3986..4801)
                     /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
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNILQFHLMLDEFF"
     primer_bind     4709..4728
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4793..4812
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5181..5769
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5670..5689
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5923..5940
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6240..6261
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6276..6281,6282..6299,6300..6306)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6314..6330
                     /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     6319..6341
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6338..6354
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6338..6354
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6531..6733
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6684..6704
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(6730..6733,1..21)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccatagg
       61 agacaccggg accgatccag cctggggatc ccgaccatgg cttcgtaccc ctgccatcaa
      121 cacgcgtctg cgttcgacca ggctgcgcgt tctcgcggcc atagcaaccg acgtacggcg
      181 ttgcgccctc gccggcagca agaagccacg gaagtccgcc tggagcagaa aatgcccacg
      241 ctactgcggg tttatataga cggtcctcac gggatgggga aaaccaccac cacgcaactg
      301 ctggtggccc tgggttcgcg cgacgatatc gtctacgtac ccgagccgat gacttactgg
      361 caggtgctgg gggcttccga gacaatcgcg aacatctaca ccacacaaca ccgcctcgac
      421 cagggtgaga tatcggccgg ggacgcggcg gtggtaatga caagcgccca gataacaatg
      481 ggcatgcctt atgccgtgac cgacgccgtt ctggctcctc atatcggggg ggaggctggg
      541 agctcacatg ccccgccccc ggccctcacc ctcatcttcg accgccatcc catcgccgcc
      601 ctcctgtgct acccggccgc gcgatacctt atgggcagca tgacccccca ggccgtgctg
      661 gcgttcgtgg ccctcatccc gccgaccttg cccggcacaa acatcgtgtt gggggccctt
      721 ccggaggaca gacacatcga ccgcctggcc aaacgccagc gccccggcga gcggcttgac
      781 ctggctatgc tggccgcgat tcgccgcgtt tacgggctgc ttgccaatac ggtgcggtat
      841 ctgcagggcg gcgggtcgtg gcgggaggat tggggacagc tttcggggac ggccgtgccg
      901 ccccagggtg ccgagcccca gagcaacgcg ggcccacgac cccatatcgg ggacacgtta
      961 tttaccctgt ttcgggcccc cgagttgctg gcccccaacg gcgacctgta caacgtgttt
     1021 gcctgggcct tggacgtctt ggccaaacgc ctccgtccca tgcacgtctt tatcctggat
     1081 tacgaccaat cgcccgccgg ctgccgggac gccctgctgc aacttacctc cgggatggtc
     1141 cagacccacg tcaccacccc cggctccata ccgacgatct gcgacctggc gcgcacgttt
     1201 gcccgggaga tgggggaggc taactgagga tcccacgtca ctattgtata ctctatatta
     1261 tactctatgt tatactctgt aatcctactc aataaacgtg tcacgcctgt gaaaccgtac
     1321 taagtctccc gtgtattctt atcaccatca ggtgacatcc tcgcccaggc tgtcaatcat
     1381 gccggtatcg attccagtag caccggcccc acgctgacaa cccactcttg cagcgttagc
     1441 agcgcccctc ttaacaagcc gacccccacc agcgtcgcgg ttactaacac tcctctcccc
     1501 gacctgcagc ccagcttgct cacattggta tggaatggac agtcaacaaa tacttaatga
     1561 atttgttata atttacaatg aaaagtttca aaatggcatt tatttctcat gatatggcaa
     1621 gtaaaaattt cacttgccag atcatgtaat aatgaatgtg ctgattttga aagccatttc
     1681 ttatgttttt agggtgagaa tccttttgaa attcaagacc attctcagga tcagcaaata
     1741 gaaggagatg aggaggatga agagaagatt gatgaaccta ttgaagaaga ggaggatgaa
     1801 gatgaagaag aagaagcaga ggaagtgggg gaagtagagg aagtggaaga agtagaggaa
     1861 gtgagagaag gaggaataga gggcgaggga aatatacagg gagtagggga aggaggggaa
     1921 gtaggggtag tgggagaagt agagggagtg ggggaagtag aggaagtaga ggaattagag
     1981 gaagagacag caaaggaaga acctgctgac ttccctgttg agcaacctga agaaaatggg
     2041 agcggaggag gttccggtgg aggtggttct ggagattaca aggatgacga cgataagggc
     2101 gattacaagg atgacgacga taagggagat tacaaggatg acgacgataa ggtttcagga
     2161 agcggagcta ctaacttcag cctgctgaag caggctggag acgtggagga gaaccctgga
     2221 cctggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     2281 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     2341 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     2401 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     2461 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     2521 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     2581 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     2641 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     2701 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     2761 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     2821 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcc
     2881 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     2941 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3001 ttctatcgcc ttcttgacga gttcttctga ttcgaacatc aaaaggagct ttacatttcg
     3061 gttctaatgt aaaaaagggt aagaaattta atagcttaaa atatgaatta acacccatgt
     3121 tgcatgcatt ccacatatta aaatttgttt tatataattt ctaaatgttt aacatttgtt
     3181 taataaaatg aaggcaaaac tattttagtt tagtttttat agctaccaga cttagatccg
     3241 ataaattgtt tgtataattt ttaagcttaa tttttattac tttattggtg ttaaggataa
     3301 caaatgtgta ttgttagtat atctattcta atcattttat cttaaatgct gctcttggga
     3361 gtgaatattc aagtgtgcat caattttttg aatacttacc ctggaagata taaactgggc
     3421 aagtattttg tgctctgtgt atttttttac tgcattagac attgaatagt aatttgcgtt
     3481 aagatacgct taaaggctct ttgtgaccat gtttcccttt gtagcaataa aatgtttttt
     3541 acgaaaactt tctccctgga ttagcagttt aaatgaaaca gagttcatca atgaaatgag
     3601 tatttaaaat aaaaatttgc cttaatgtat cagttcagct cacaagtatt ttaagatgat
     3661 tgagaagact tgaattaaag aaaaaaaaat tctcaatcct tggaaagtcc tctccactga
     3721 ctgtagcctc caattcactg gccgtcgttc actggccgtc gttttacaac gtcgtgactg
     3781 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     3841 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     3901 cgaatgcgat ttattcaaca aagccgccgt cccgtcaagt cagcgtaatg ctctgccagt
     3961 gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca
     4021 atatattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     4081 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     4141 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     4201 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt
     4261 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     4321 ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa
     4381 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     4441 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga
     4501 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     4561 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     4621 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
     4681 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     4741 catccatgtt ggaatttaat cgcggcttcg agcaagacgt ttcccgttga atatggctca
     4801 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat
     4861 ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt tgttgaataa
     4921 atcgaacttt tgctgagttg aaggatcaga tcacgcatct tcccgacaac gcagaccgtt
     4981 ccgtggcaaa gcaaaagttc aaaatcacca actggtccac ctacaacaaa gctctcatca
     5041 accgtggctc cctcactttc tggctggatg atggggcgat tcaggcctgg tatgagtcag
     5101 caacaccttc ttcacgaggc agacctctcg acggagttcc actgagcgtc agaccccgta
     5161 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5221 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5281 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5341 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5401 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5461 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5521 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5581 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5641 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5701 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5761 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5821 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     5881 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     5941 gaagcggaag aagctcgcac attcagcagc gtttttcagc gcgttttcga tcaacgtttc
     6001 aatgttggta tcaacaccag gtttaacttt gaacttatcg gcactgacgg ttactgattt
     6061 tgaacttttg ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg atctgatcct
     6121 tcaactcagc aaaagttccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6181 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     6241 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     6301 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     6361 tacgccaagc ttgcatgcct gcaggtcgac tctagaggat ccccgggtac cgagctcgaa
     6421 ttagcttgca tgcctgcagg tcaattccct ggcattatgc ccagtacatg accttatggg
     6481 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     6541 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt caagtctcca
     6601 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     6661 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     6721 tataagcaga gct
//
