LOCUS       sequence-245630-        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             558..581
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             585..608
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             612..635
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             651..707
                     /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             720..1514
                     /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(774..793)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1384..1403
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2235..2251)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2244..2266)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2470..3285)
                     /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     3193..3212
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3277..3296
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3665..4253
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4154..4173
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4407..4424
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4724..4745
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4760..4765,4766..4783,4784..4790)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4798..4814
                     /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     4803..4825
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4822..4838
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4822..4838
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5015..5217
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5168..5188
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5214..5238
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5314..6444
                     /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 tgctcacatt ggtatggaat ggacagtcaa caaatactta atgaatttgt tataatttac
       61 aatgaaaagt ttcaaaatgg catttatttc tcatgatatg gcaagtaaaa atttcacttg
      121 ccagatcatg taataatgaa tgtgctgatt ttgaaagcca tttcttatgt ttttagggtg
      181 agaatccttt tgaaattcaa gaccattctc aggatcagca aatagaagga gatgaggagg
      241 atgaagagaa gattgatgaa cctattgaag aagaggagga tgaagatgaa gaagaagaag
      301 cagaggaagt gggggaagta gaggaagtgg aagaagtaga ggaagtgaga gaaggaggaa
      361 tagagggcga gggaaatata cagggagtag gggaaggagg ggaagtaggg gtagtgggag
      421 aagtagaggg agtgggggaa gtagaggaag tagaggaatt agaggaagag acagcaaagg
      481 aagaacctgc tgacttccct gttgagcaac ctgaagaaaa tgggagcgga ggaggttccg
      541 gtggaggtgg ttctggagat tacaaggatg acgacgataa gggcgattac aaggatgacg
      601 acgataaggg agattacaag gatgacgacg ataaggtttc aggaagcgga gctactaact
      661 tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctgga tcgtttcgca
      721 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
      781 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
      841 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
      901 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
      961 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     1021 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     1081 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     1141 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     1201 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     1261 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     1321 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggccgaccgc tatcaggaca
     1381 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     1441 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     1501 acgagttctt ctgattcgaa catcaaaagg agctttacat ttcggttcta atgtaaaaaa
     1561 gggtaagaaa tttaatagct taaaatatga attaacaccc atgttgcatg cattccacat
     1621 attaaaattt gttttatata atttctaaat gtttaacatt tgtttaataa aatgaaggca
     1681 aaactatttt agtttagttt ttatagctac cagacttaga tccgataaat tgtttgtata
     1741 atttttaagc ttaattttta ttactttatt ggtgttaagg ataacaaatg tgtattgtta
     1801 gtatatctat tctaatcatt ttatcttaaa tgctgctctt gggagtgaat attcaagtgt
     1861 gcatcaattt tttgaatact taccctggaa gatataaact gggcaagtat tttgtgctct
     1921 gtgtattttt ttactgcatt agacattgaa tagtaatttg cgttaagata cgcttaaagg
     1981 ctctttgtga ccatgtttcc ctttgtagca ataaaatgtt ttttacgaaa actttctccc
     2041 tggattagca gtttaaatga aacagagttc atcaatgaaa tgagtattta aaataaaaat
     2101 ttgccttaat gtatcagttc agctcacaag tattttaaga tgattgagaa gacttgaatt
     2161 aaagaaaaaa aaattctcaa tccttggaaa gtcctctcca ctgactgtag cctccaattc
     2221 actggccgtc gttcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     2281 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     2341 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg cgatttattc
     2401 aacaaagccg ccgtcccgtc aagtcagcgt aatgctctgc cagtgttaca accaattaac
     2461 caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     2521 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     2581 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     2641 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     2701 agtgacgact gaatccggtg agaatggcaa aagcttatgc atttctttcc agacttgttc
     2761 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     2821 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     2881 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     2941 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     3001 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     3061 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     3121 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     3181 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     3241 taatcgcggc ttcgagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     3301 actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat cttgtgcaat
     3361 gtaacatcag agattttgag acacaacgtg gctttgttga ataaatcgaa cttttgctga
     3421 gttgaaggat cagatcacgc atcttcccga caacgcagac cgttccgtgg caaagcaaaa
     3481 gttcaaaatc accaactggt ccacctacaa caaagctctc atcaaccgtg gctccctcac
     3541 tttctggctg gatgatgggg cgattcaggc ctggtatgag tcagcaacac cttcttcacg
     3601 aggcagacct ctcgacggag ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3661 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3721 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3781 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3841 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3901 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3961 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4021 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4081 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4141 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4201 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4261 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4321 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4381 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagaagctc
     4441 gcacattcag cagcgttttt cagcgcgttt tcgatcaacg tttcaatgtt ggtatcaaca
     4501 ccaggtttaa ctttgaactt atcggcactg acggttactg attttgaact tttgctttgc
     4561 cacggaacgg tctgcgttgt cgggaagatg cgtgatctga tccttcaact cagcaaaagt
     4621 tcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     4681 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     4741 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     4801 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcttgcat
     4861 gcctgcaggt cgactctaga ggatccccgg gtaccgagct cgaattagct tgcatgcctg
     4921 caggtcaatt ccctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     4981 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     5041 tgggcgtgga tagcggtttg actcacgggg atttcaagtc tccaccccat tgacgtcaat
     5101 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     5161 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
     5221 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccataggaga
     5281 caccgggacc gatccagcct ggggatcccg accatggctt cgtacccctg ccatcaacac
     5341 gcgtctgcgt tcgaccaggc tgcgcgttct cgcggccata gcaaccgacg tacggcgttg
     5401 cgccctcgcc ggcagcaaga agccacggaa gtccgcctgg agcagaaaat gcccacgcta
     5461 ctgcgggttt atatagacgg tcctcacggg atggggaaaa ccaccaccac gcaactgctg
     5521 gtggccctgg gttcgcgcga cgatatcgtc tacgtacccg agccgatgac ttactggcag
     5581 gtgctggggg cttccgagac aatcgcgaac atctacacca cacaacaccg cctcgaccag
     5641 ggtgagatat cggccgggga cgcggcggtg gtaatgacaa gcgcccagat aacaatgggc
     5701 atgccttatg ccgtgaccga cgccgttctg gctcctcata tcggggggga ggctgggagc
     5761 tcacatgccc cgcccccggc cctcaccctc atcttcgacc gccatcccat cgccgccctc
     5821 ctgtgctacc cggccgcgcg ataccttatg ggcagcatga ccccccaggc cgtgctggcg
     5881 ttcgtggccc tcatcccgcc gaccttgccc ggcacaaaca tcgtgttggg ggcccttccg
     5941 gaggacagac acatcgaccg cctggccaaa cgccagcgcc ccggcgagcg gcttgacctg
     6001 gctatgctgg ccgcgattcg ccgcgtttac gggctgcttg ccaatacggt gcggtatctg
     6061 cagggcggcg ggtcgtggcg ggaggattgg ggacagcttt cggggacggc cgtgccgccc
     6121 cagggtgccg agccccagag caacgcgggc ccacgacccc atatcgggga cacgttattt
     6181 accctgtttc gggcccccga gttgctggcc cccaacggcg acctgtacaa cgtgtttgcc
     6241 tgggccttgg acgtcttggc caaacgcctc cgtcccatgc acgtctttat cctggattac
     6301 gaccaatcgc ccgccggctg ccgggacgcc ctgctgcaac ttacctccgg gatggtccag
     6361 acccacgtca ccacccccgg ctccataccg acgatctgcg acctggcgcg cacgtttgcc
     6421 cgggagatgg gggaggctaa ctgaggatcc cacgtcacta ttgtatactc tatattatac
     6481 tctatgttat actctgtaat cctactcaat aaacgtgtca cgcctgtgaa accgtactaa
     6541 gtctcccgtg tattcttatc accatcaggt gacatcctcg cccaggctgt caatcatgcc
     6601 ggtatcgatt ccagtagcac cggccccacg ctgacaaccc actcttgcag cgttagcagc
     6661 gcccctctta acaagccgac ccccaccagc gtcgcggtta ctaacactcc tctccccgac
     6721 ctgcagccca gct
//
