LOCUS       sequence-271139-        6939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human ZNF264_iso1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6939)
  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 6939)
  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_ZNF264_iso1
FEATURES             Location/Qualifiers
     source          1..6939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..203
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     154..174
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     200..224
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             300..1430
                     /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             2433..2456
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2460..2483
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2487..2510
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2526..2582
                     /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             2595..3389
                     /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(2649..2668)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3259..3278
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4160..4176)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4169..4191)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(4395..5210)
                     /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     5118..5137
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5202..5221
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5590..6178
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6079..6098
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6332..6349
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6649..6670
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6685..6690,6691..6708,6709..6715)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6723..6739
                     /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     6728..6750
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6747..6763
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6747..6763
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      181 gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
      241 cacgctgttt tgacctccat aggagacacc gggaccgatc cagcctgggg atcccgacca
      301 tggcttcgta cccctgccat caacacgcgt ctgcgttcga ccaggctgcg cgttctcgcg
      361 gccatagcaa ccgacgtacg gcgttgcgcc ctcgccggca gcaagaagcc acggaagtcc
      421 gcctggagca gaaaatgccc acgctactgc gggtttatat agacggtcct cacgggatgg
      481 ggaaaaccac caccacgcaa ctgctggtgg ccctgggttc gcgcgacgat atcgtctacg
      541 tacccgagcc gatgacttac tggcaggtgc tgggggcttc cgagacaatc gcgaacatct
      601 acaccacaca acaccgcctc gaccagggtg agatatcggc cggggacgcg gcggtggtaa
      661 tgacaagcgc ccagataaca atgggcatgc cttatgccgt gaccgacgcc gttctggctc
      721 ctcatatcgg gggggaggct gggagctcac atgccccgcc cccggccctc accctcatct
      781 tcgaccgcca tcccatcgcc gccctcctgt gctacccggc cgcgcgatac cttatgggca
      841 gcatgacccc ccaggccgtg ctggcgttcg tggccctcat cccgccgacc ttgcccggca
      901 caaacatcgt gttgggggcc cttccggagg acagacacat cgaccgcctg gccaaacgcc
      961 agcgccccgg cgagcggctt gacctggcta tgctggccgc gattcgccgc gtttacgggc
     1021 tgcttgccaa tacggtgcgg tatctgcagg gcggcgggtc gtggcgggag gattggggac
     1081 agctttcggg gacggccgtg ccgccccagg gtgccgagcc ccagagcaac gcgggcccac
     1141 gaccccatat cggggacacg ttatttaccc tgtttcgggc ccccgagttg ctggccccca
     1201 acggcgacct gtacaacgtg tttgcctggg ccttggacgt cttggccaaa cgcctccgtc
     1261 ccatgcacgt ctttatcctg gattacgacc aatcgcccgc cggctgccgg gacgccctgc
     1321 tgcaacttac ctccgggatg gtccagaccc acgtcaccac ccccggctcc ataccgacga
     1381 tctgcgacct ggcgcgcacg tttgcccggg agatggggga ggctaactga ggatcccacg
     1441 tcactattgt atactctata ttatactcta tgttatactc tgtaatccta ctcaataaac
     1501 gtgtcacgcc tgtgaaaccg tactaagtct cccgtgtatt cttatcacca tcaggtgaca
     1561 tcctcgccca ggctgtcaat catgccggta tcgattccag tagcaccggc cccacgctga
     1621 caacccactc ttgcagcgtt agcagcgccc ctcttaacaa gccgaccccc accagcgtcg
     1681 cggttactaa cactcctctc cccgacctgc agcccagctt cgagtgtggg aaggctttca
     1741 accaccgatc ctacctcaag aggcaccagc ggattcacac tggggagaag cccttcgtgt
     1801 gcagtgaatg tggaaaggcc ttcacccact gctctacttt tatcttgcat aaaagggccc
     1861 acactggaga aaagcctttc gagtgcaaag agtgtgggaa agcctttagc aatcggaagg
     1921 acctcattcg ccacttcagc atccacactg gagagaagcc ctatgagtgc gtggagtgtg
     1981 gaaaggcctt cacccgcatg tcgggcctca cgaggcacaa gcggattcat agtggagaga
     2041 agccctatga atgtgttgag tgtgggaaat cgttttgctg gagcacaaac ctcattcgac
     2101 atgccattat ccacactgga gagaagccct ataaatgtag tgaatgtgga aaggccttca
     2161 gtcgcagctc gtccctcact cagcatcaaa ggatgcatac tgggaaaaat cccatcagtg
     2221 taacagatgt gggaagacct tttacaagtg gacaaacctc agttaccctt cgagaacttc
     2281 ttttagggaa ggactttttg aatgtaacca ctgaggcaaa tattttgcca gaggaaacat
     2341 cttcctctgc atctgatcaa ccataccaaa gagaaacccc acaagtgtct tcactgggga
     2401 gcggaggagg ttccggtgga ggtggttctg gagattacaa ggatgacgac gataagggcg
     2461 attacaagga tgacgacgat aagggagatt acaaggatga cgacgataag gtttcaggaa
     2521 gcggagctac taacttcagc ctgctgaagc aggctggaga cgtggaggag aaccctggac
     2581 ctggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     2641 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     2701 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     2761 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     2821 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     2881 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     2941 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     3001 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     3061 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     3121 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     3181 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggccg
     3241 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     3301 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     3361 tctatcgcct tcttgacgag ttcttctgat tcgaacatcg ttgctgaata ttacttgtca
     3421 tctgaagagt catattagaa attcgttcag tctagagcct tattctccat ctgataattt
     3481 atcctggaga gagacccagt ggttattgtg cacataggag aaccttcagc tgcatctttc
     3541 tccttagttt acagtgcaat tttatctcag gaattatttt taaaaggagg aggggacata
     3601 gaaaaaatga aatgcaagca cacatctttt caggcttctc tgccaagcct atggcgcttt
     3661 gtcatggatt tcttagtgta tttgggggaa gggaaatgtt tcaaggtaaa gaaccttgac
     3721 cctttatgtg cttgtatgta catttattgc tatccagtgt cagaacagtt agtttaggaa
     3781 aagtatgcaa actttaatcg cacatcttct gtattccaca atagtactta ctcttgagaa
     3841 gcataacttt atgacaactt agggggcttg agccatgaaa tactcacgtt taagtcagta
     3901 aggatacaca tgttaacatt caggcttttg tcttgatgcc cgtcttttgg tttacgtcat
     3961 cattgtagcc atatggtaaa tttttatttg ttaaattttt aaaaattact tagctcaaaa
     4021 tgtcagtggt aaatttttaa attgagtaaa gtgattcttc tttgctctta tttaaaatcg
     4081 acagcatttc tagttccttt gacattcctt ggaaagtcct ctccactgac tgtagcctcc
     4141 aattcactgg ccgtcgttca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     4201 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     4261 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgcgatt
     4321 tattcaacaa agccgccgtc ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa
     4381 ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tatattcata
     4441 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     4501 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     4561 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     4621 ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact
     4681 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     4741 ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta
     4801 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     4861 cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg
     4921 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     4981 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     5041 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca
     5101 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     5161 gaatttaatc gcggcttcga gcaagacgtt tcccgttgaa tatggctcat aacacccctt
     5221 gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt
     5281 gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt
     5341 gctgagttga aggatcagat cacgcatctt cccgacaacg cagaccgttc cgtggcaaag
     5401 caaaagttca aaatcaccaa ctggtccacc tacaacaaag ctctcatcaa ccgtggctcc
     5461 ctcactttct ggctggatga tggggcgatt caggcctggt atgagtcagc aacaccttct
     5521 tcacgaggca gacctctcga cggagttcca ctgagcgtca gaccccgtag aaaagatcaa
     5581 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5641 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5701 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5761 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5821 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5881 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5941 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6001 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6061 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6121 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6181 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6241 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     6301 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     6361 agctcgcaca ttcagcagcg tttttcagcg cgttttcgat caacgtttca atgttggtat
     6421 caacaccagg tttaactttg aacttatcgg cactgacggt tactgatttt gaacttttgc
     6481 tttgccacgg aacggtctgc gttgtcggga agatgcgtga tctgatcctt caactcagca
     6541 aaagttccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     6601 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     6661 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     6721 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
     6781 tgcatgcctg caggtcgact ctagaggatc cccgggtacc gagctcgaat tagcttgcat
     6841 gcctgcaggt caattccctg gcattatgcc cagtacatga ccttatggga ctttcctact
     6901 tggcagtaca tctacgtatt agtcatcgct attaccatg
//
