LOCUS       sequence-269898-        6933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human ONECUT2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6933)
  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 6933)
  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_ONECUT2
FEATURES             Location/Qualifiers
     source          1..6933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             960..983
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             987..1010
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1014..1037
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1053..1109
                     /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             1122..1916
                     /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(1176..1195)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1786..1805
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2650..2666)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2659..2681)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2885..3700)
                     /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     3608..3627
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3692..3711
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4080..4668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4569..4588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4822..4839
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5139..5160
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5175..5180,5181..5198,5199..5205)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5213..5229
                     /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     5218..5240
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5237..5253
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5237..5253
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5430..5632
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5583..5603
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5629..5653
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5729..6859
                     /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 cacgcctgtg aaaccgtact aagtctcccg tgtattctta tcaccatcag gtgacatcct
       61 cgcccaggct gtcaatcatg ccggtatcga ttccagtagc accggcccca cgctgacaac
      121 ccactcttgc agcgttagca gcgcccctct taacaagccg acccccacca gcgtcgcggt
      181 tactaacact cctctccccg acctgcagcc cagctaattt caactcagct gtaaacaatt
      241 cgagagggtg aatatgaaag aggaactgcc tcaccaggct tcccccaccc tggcgtctgg
      301 agtcctccct gccttcgtgg cacctgattt cagagataaa gaatttcaag gatatgctag
      361 agcagggagg atgtaaaagg gaaagacaac tgcagttagc tcaaaaccag actgtgtgaa
      421 acagccctgc tgctttgtcc tttctcccat cccccactca ctcactgatg ccttccatca
      481 cccggagatt acagggaagg ctggttattg acagctgggc agtttgcaca ataaagttta
      541 acggtttaca tctctttgta caactttgtc aatgttttcc atggatcacc ttctcaggtg
      601 agcccactga ctcctctcct tcttctcttt cccttcaagc gtgcaaacgc aaagagcaag
      661 aaccaaacaa agacaggaac aattcccaga agaagtcccg cctggtgttc actgacctcc
      721 aacgccgaac actcttcgcc atcttcaagg agaacaaacg cccgtcaaag gagatgcaga
      781 tcaccatttc ccagcagctg ggcctggagc tcacaaccgt cagcaacttc ttcatgaacg
      841 cccggcgccg cagcctggag aagtggcaag acgatctgag cacagggggc tcctcgtcca
      901 cctccagcac gtgtaccaaa gcagggagcg gaggaggttc cggtggaggt ggttctggag
      961 attacaagga tgacgacgat aagggcgatt acaaggatga cgacgataag ggagattaca
     1021 aggatgacga cgataaggtt tcaggaagcg gagctactaa cttcagcctg ctgaagcagg
     1081 ctggagacgt ggaggagaac cctggacctg gatcgtttcg catgattgaa caagatggat
     1141 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     1201 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     1261 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     1321 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     1381 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     1441 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     1501 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     1561 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     1621 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     1681 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     1741 tcgactgtgg ccggctgggt gtggccgacc gctatcagga catagcgttg gctacccgtg
     1801 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     1861 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgattcg
     1921 aacatccttg ggcacaagtc acctccaaat gaggacaaca gataccaaaa gaaaacaaag
     1981 gaaaaagaca ccggattcct agctggggcc cttcactggt gatttgaaag cacaattctc
     2041 ttgcaaagaa acttatattc tagctgtaat cataggccag gtgttcttct tttgttttta
     2101 atggctatgg agtccaagtg caagctgaaa aattaatctc ttagaaccag acactgttct
     2161 ctgagcatgc taagcatccc agaaacccaa atggggcctt cctggagcga gttaattcca
     2221 gtatggtgtc aaccaagctc gggattgctt aaaatatcat ccatcccact tcaggtcctg
     2281 tcagcttctt gcagtcagag ttcctatgag taacaatagg agtttggcct atgtaaggac
     2341 tctgagttta ggcttccaag atacaacaat aagagaagaa tctagcaacg agaatgacct
     2401 catttgcttt ccacatgctt agcctcatta taccatgtta tgtccaagtt cacagccaca
     2461 acatcagaat ggtaattact gagcacaagt tttaaatatg gacgttaaaa aaaaaaatcc
     2521 aaggacctgt ttttccaacc cagacatctt ttcattgaat gatttagaaa gctttaagtt
     2581 gatccagctt acaatttctt ggaaagtcct ctccactgac tgtagcctcc aattcactgg
     2641 ccgtcgttca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     2701 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     2761 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgcgatt tattcaacaa
     2821 agccgccgtc ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt
     2881 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tatattcata tcaggattat
     2941 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     3001 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     3061 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     3121 cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag
     3181 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     3241 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     3301 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     3361 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     3421 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     3481 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     3541 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     3601 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     3661 gcggcttcga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     3721 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     3781 atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt gctgagttga
     3841 aggatcagat cacgcatctt cccgacaacg cagaccgttc cgtggcaaag caaaagttca
     3901 aaatcaccaa ctggtccacc tacaacaaag ctctcatcaa ccgtggctcc ctcactttct
     3961 ggctggatga tggggcgatt caggcctggt atgagtcagc aacaccttct tcacgaggca
     4021 gacctctcga cggagttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4081 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4141 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4201 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4261 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4321 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4381 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4441 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4501 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4561 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4621 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4681 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4741 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4801 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga agctcgcaca
     4861 ttcagcagcg tttttcagcg cgttttcgat caacgtttca atgttggtat caacaccagg
     4921 tttaactttg aacttatcgg cactgacggt tactgatttt gaacttttgc tttgccacgg
     4981 aacggtctgc gttgtcggga agatgcgtga tctgatcctt caactcagca aaagttccca
     5041 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     5101 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     5161 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     5221 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct tgcatgcctg
     5281 caggtcgact ctagaggatc cccgggtacc gagctcgaat tagcttgcat gcctgcaggt
     5341 caattccctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     5401 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     5461 gtggatagcg gtttgactca cggggatttc aagtctccac cccattgacg tcaatgggag
     5521 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     5581 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
     5641 gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata ggagacaccg
     5701 ggaccgatcc agcctgggga tcccgaccat ggcttcgtac ccctgccatc aacacgcgtc
     5761 tgcgttcgac caggctgcgc gttctcgcgg ccatagcaac cgacgtacgg cgttgcgccc
     5821 tcgccggcag caagaagcca cggaagtccg cctggagcag aaaatgccca cgctactgcg
     5881 ggtttatata gacggtcctc acgggatggg gaaaaccacc accacgcaac tgctggtggc
     5941 cctgggttcg cgcgacgata tcgtctacgt acccgagccg atgacttact ggcaggtgct
     6001 gggggcttcc gagacaatcg cgaacatcta caccacacaa caccgcctcg accagggtga
     6061 gatatcggcc ggggacgcgg cggtggtaat gacaagcgcc cagataacaa tgggcatgcc
     6121 ttatgccgtg accgacgccg ttctggctcc tcatatcggg ggggaggctg ggagctcaca
     6181 tgccccgccc ccggccctca ccctcatctt cgaccgccat cccatcgccg ccctcctgtg
     6241 ctacccggcc gcgcgatacc ttatgggcag catgaccccc caggccgtgc tggcgttcgt
     6301 ggccctcatc ccgccgacct tgcccggcac aaacatcgtg ttgggggccc ttccggagga
     6361 cagacacatc gaccgcctgg ccaaacgcca gcgccccggc gagcggcttg acctggctat
     6421 gctggccgcg attcgccgcg tttacgggct gcttgccaat acggtgcggt atctgcaggg
     6481 cggcgggtcg tggcgggagg attggggaca gctttcgggg acggccgtgc cgccccaggg
     6541 tgccgagccc cagagcaacg cgggcccacg accccatatc ggggacacgt tatttaccct
     6601 gtttcgggcc cccgagttgc tggcccccaa cggcgacctg tacaacgtgt ttgcctgggc
     6661 cttggacgtc ttggccaaac gcctccgtcc catgcacgtc tttatcctgg attacgacca
     6721 atcgcccgcc ggctgccggg acgccctgct gcaacttacc tccgggatgg tccagaccca
     6781 cgtcaccacc cccggctcca taccgacgat ctgcgacctg gcgcgcacgt ttgcccggga
     6841 gatgggggag gctaactgag gatcccacgt cactattgta tactctatat tatactctat
     6901 gttatactct gtaatcctac tcaataaacg tgt
//
