LOCUS       sequence-199975-        8492 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human RXRB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8492)
  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 8492)
  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_RXRB
FEATURES             Location/Qualifiers
     source          1..8492
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(216..233)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(387..975)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(467..486)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1344..1363)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1355..2170
                     /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     complement(1428..1447)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3712..3734
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3727..3743
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(4606..5400)
                     /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(4717..4736)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5327..5346
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(5413..5469)
                     /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             complement(5485..5508)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(5512..5535)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(5539..5562)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(6688..7818)
                     /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"
     primer_bind     complement(7894..7918)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(7915..8117)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(7944..7964)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(8294..8310)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8294..8310)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8307..8329)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8318..8334
                     /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)."
     promoter        complement(join(8342..8348,8349..8366,8367..8372))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8387..8408
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ggcggtttgc gtattgggaa cttttgctga gttgaaggat cagatcacgc atcttcccga
       61 caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc agtaaccgtc agtgccgata
      121 agttcaaagt taaacctggt gttgatacca acattgaaac gttgatcgaa aacgcgctga
      181 aaaacgctgc tgaatgtgcg agcttcttcc gcttcctcgc tcactgactc gctgcgctcg
      241 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
      301 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
      361 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
      421 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
      481 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
      541 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
      601 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
      661 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
      721 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
      781 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
      841 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
      901 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
      961 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     1021 tccgtcgaga ggtctgcctc gtgaagaagg tgttgctgac tcataccagg cctgaatcgc
     1081 cccatcatcc agccagaaag tgagggagcc acggttgatg agagctttgt tgtaggtgga
     1141 ccagttggtg attttgaact tttgctttgc cacggaacgg tctgcgttgt cgggaagatg
     1201 cgtgatctga tccttcaact cagcaaaagt tcgatttatt caacaaagcc acgttgtgtc
     1261 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     1321 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     1381 ttgctcgaag ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     1441 tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc
     1501 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     1561 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     1621 tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt
     1681 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     1741 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     1801 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     1861 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataagcttt tgccattctc
     1921 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     1981 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     2041 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     2101 aaaatatggt attgataatc ctgatatgaa tatattgcag tttcatttga tgctcgatga
     2161 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcactg agagatcccc
     2221 tcataatttc cccaaagcgt aaccatgtgt gaataaattt tgagctagta gggttgcagc
     2281 cacgagtaag tcttcccttg ttattgtgta gccagaatgc cgcaaaactt ccatgcctaa
     2341 gcgaactgtt gagagtacgt ttcgatttct gactgtgtta gcctggaagt gcttgtccca
     2401 accttgtttc tgagcatgaa cgcccgcaag ccaacatgtt agttgaagca tcagggcgat
     2461 tagcagcatg atatcaaaac gctctgagct gctcgttcgg ctatggcgta ggcctagtcc
     2521 gtaggcagga cttttcaagt ctcggaaggt ttcttcaatc tgcattcgct tcgaatagat
     2581 attaacaagt tgtttgggtg ttcgaatttc aacaggtaag ttagttgcta gaacccatgg
     2641 ctcctttgcc gacgctgagt agattttagg tgacgggtgg tgacaatgag tccgtgtcga
     2701 gcgctgattt tttcggcctt tagagcgaga tttatacaat agaatttggc atgagattgg
     2761 attgctttta gtcagcctct tatagcctaa agtctttgag tgactagatg acatatcatg
     2821 taagttgctg ataggtttcc agttttccgc tcctaggtct gcatattgta cttttcctct
     2881 tactcgactt aaccagtacc aacccagctt ctcaacggat ttataccatg gcactttaaa
     2941 gccagcatca ctgacaatga gcggtgtggt gttactcggt agaatgctcg caaggtcggc
     3001 tagaaattgg tcatgagctt tctttgaaca ttgctctgaa agcgggaacg ctttctcata
     3061 aagagtaaca gaacgaccgt gtagtgcgac tgaagctcgc aataccataa gtcgtttttg
     3121 ctcacgaata tcagaccagt caacaagtac aatgggcatc gtattgcccg aacagataaa
     3181 gctagcatgc caacggtata cagcgagtcg ctctttgtgg aggtgacgat tacctaacaa
     3241 tcggtcgatt cgtttgatgt tatgttttgt tctcgctttg gttggcaggt tacggccaag
     3301 ttcggtaaga gtgagagttt tacagtcaag taatgcgtgg caagccaacg ttaagctgtt
     3361 gagtcgtttt aagtgtaatt cggggcagaa ttggtaaaga gagtcgtgta aaatatcgag
     3421 ttcgcacatc ttgttgtctg attattgatt tttcgcgaaa ccatttgatc atatgacaag
     3481 atgtgtatcc accttaactt aatgattttt accaaaatca ttaggggatt catcagggca
     3541 gagcattacg ctgacttgac gggacggcgg ctttgttgaa taaatcgcat tcgccattca
     3601 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     3661 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     3721 gacgttgtaa aacgacggcc agtgaacgac ggccagtgaa ttggaggcta cagtcagtgg
     3781 agaggacttt ccaagtagac aggaagagat agggggggcg gcgggaagct gggaaggcta
     3841 gtgcttggag agccctaggg acaggccatt tcagggccct gcctttccca aacacccacc
     3901 tccaccactg gcatttctta gtcaacctgg gaaagtacag tacttctttg agtctaactg
     3961 caagtctcta tcctcacagg aaattaaaaa tagcagatcg gttcctacat ctccaccagc
     4021 cccttcacca ccaccaccac cttttttata tttcagtctg actgcagaag gaggtgaagt
     4081 gtaaaaagag actctggaca gtgacagggc ccctccctct tccagagagg cccccatctg
     4141 ccaggtttga gaggaggaag gcctgtcagg gccctactct catgtccatc agcttgggag
     4201 gcctgccccc cagtatccac ctctggggga gttccccatt tccactcttc agatgggaag
     4261 caaaatgagg caagatgaga aggaagcaag gtcctggagg caaggccagt gctttgtgct
     4321 gggggaagga cagagggtga gaaatcaccc caaatcatgg gagaacccga caaattcaga
     4381 gactcaaggc caccgaagag agacaaccag tcctcacagg tatctggggt cccttccaac
     4441 ttgggatatc aagcagatcc cttggagggt ttatgttctt ggttctgccc tgtacttctc
     4501 accccatcaa ggttctggga acatggcccc ccaccctgcc ccagggcttg gagtccctct
     4561 tggatgtgtg ctcctccagt gtgagaagca ccacggatgt tcgaatcaga agaactcgtc
     4621 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag
     4681 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat
     4741 gtcctgatag cggtcggcca cacccagccg gccacagtcg atgaatccag aaaagcggcc
     4801 attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc
     4861 gtcgggcatg cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc
     4921 ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat
     4981 gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg
     5041 cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc
     5101 ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag
     5161 cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg
     5221 cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc
     5281 tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc
     5341 gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat
     5401 gcgaaacgat ccaggtccag ggttctcctc cacgtctcca gcctgcttca gcaggctgaa
     5461 gttagtagct ccgcttcctg aaaccttatc gtcgtcatcc ttgtaatctc ccttatcgtc
     5521 gtcatccttg taatcgccct tatcgtcgtc atccttgtaa tctccagaac cacctccacc
     5581 ggaacctcct ccgctcccgg ccagttgatg gggagcctca agcatctcca tgaggaaggt
     5641 gtcgatgggg gtgtcaccaa tgagcttgaa gaaaaacaga tgctctagac acttaaggcc
     5701 aatggaccgg agggcaggaa gacgtagcag cagcttggca aacctggggt ggaggtggga
     5761 gaaggggatt gagagctgga agcacacggg ccctgaacac atcctcatag cactccccac
     5821 ccccaaggga gcctcagtgc cccccagccc catctcaccg tccctgctgc tcagggtact
     5881 tctgtttgca gtaggtctcc agtgatgcat acactttctc ccgcaggacc tccacctcac
     5941 tagggttgga gaggcccttg gcatctggga tggcagggaa gagaggagga agagaaatga
     6001 agacaaacca aatcaggatg gccatgcaga tgtgagccac aggatgcccc ttttgggctg
     6061 cacttgcttg ccctttacca gaggcctggc aagggaagca gggcccactg ggtttgtggg
     6121 atggatccgt ggatgtgggt ttttcctcgg ccagttggga gatttccagg ttgagggtct
     6181 tactgagggg gatagctggg taacttagga gtctcggaga agaggaggct ccaaggttgc
     6241 cttggccttg agagacaaag gtaatcctcc tcttacctgg attaaacaga atgattgccc
     6301 tcaggcagcc aagctctgtc ttgtccatcc tcatgtcacg cattttggac actagctctg
     6361 tcagcaccct ggagagggac ctgcaggtca ctcaaaggag ctgggctgca ggtcggggag
     6421 aggagtgtta gtaaccgcga cgctggtggg ggtcggcttg ttaagagggg cgctgctaac
     6481 gctgcaagag tgggttgtca gcgtggggcc ggtgctactg gaatcgatac cggcatgatt
     6541 gacagcctgg gcgaggatgt cacctgatgg tgataagaat acacgggaga cttagtacgg
     6601 tttcacaggc gtgacacgtt tattgagtag gattacagag tataacatag agtataatat
     6661 agagtataca atagtgacgt gggatcctca gttagcctcc cccatctccc gggcaaacgt
     6721 gcgcgccagg tcgcagatcg tcggtatgga gccgggggtg gtgacgtggg tctggaccat
     6781 cccggaggta agttgcagca gggcgtcccg gcagccggcg ggcgattggt cgtaatccag
     6841 gataaagacg tgcatgggac ggaggcgttt ggccaagacg tccaaggccc aggcaaacac
     6901 gttgtacagg tcgccgttgg gggccagcaa ctcgggggcc cgaaacaggg taaataacgt
     6961 gtccccgata tggggtcgtg ggcccgcgtt gctctggggc tcggcaccct ggggcggcac
     7021 ggccgtcccc gaaagctgtc cccaatcctc ccgccacgac ccgccgccct gcagataccg
     7081 caccgtattg gcaagcagcc cgtaaacgcg gcgaatcgcg gccagcatag ccaggtcaag
     7141 ccgctcgccg gggcgctggc gtttggccag gcggtcgatg tgtctgtcct ccggaagggc
     7201 ccccaacacg atgtttgtgc cgggcaaggt cggcgggatg agggccacga acgccagcac
     7261 ggcctggggg gtcatgctgc ccataaggta tcgcgcggcc gggtagcaca ggagggcggc
     7321 gatgggatgg cggtcgaaga tgagggtgag ggccgggggc ggggcatgtg agctcccagc
     7381 ctcccccccg atatgaggag ccagaacggc gtcggtcacg gcataaggca tgcccattgt
     7441 tatctgggcg cttgtcatta ccaccgccgc gtccccggcc gatatctcac cctggtcgag
     7501 gcggtgttgt gtggtgtaga tgttcgcgat tgtctcggaa gcccccagca cctgccagta
     7561 agtcatcggc tcgggtacgt agacgatatc gtcgcgcgaa cccagggcca ccagcagttg
     7621 cgtggtggtg gttttcccca tcccgtgagg accgtctata taaacccgca gtagcgtggg
     7681 cattttctgc tccaggcgga cttccgtggc ttcttgctgc cggcgagggc gcaacgccgt
     7741 acgtcggttg ctatggccgc gagaacgcgc agcctggtcg aacgcagacg cgtgttgatg
     7801 gcaggggtac gaagccatgg tcgggatccc caggctggat cggtcccggt gtctcctatg
     7861 gaggtcaaaa cagcgtggat ggcgtctcca ggcgatctga cggttcacta aacgagctct
     7921 gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt
     7981 gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc
     8041 aatggggtgg agacttgaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac
     8101 tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
     8161 ggaaagtccc ataaggtcat gtactgggca taatgccagg gaattgacct gcaggcatgc
     8221 aagctaattc gagctcggta cccggggatc ctctagagtc gacctgcagg catgcaagct
     8281 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     8341 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     8401 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     8461 tgcattaatg aatcggccaa cgcgcgggga ga
//
