LOCUS       sequence-171033-        6972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human HOMEZ.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6972)
  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 6972)
  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_HOMEZ
FEATURES             Location/Qualifiers
     source          1..6972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      65..653
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     554..573
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     807..824
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1124..1145
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1160..1165,1166..1183,1184..1190)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1198..1214
                     /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     1203..1225
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1222..1238
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1222..1238
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1415..1617
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1568..1588
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1614..1638
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1714..2844
                     /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             complement(3853..3873)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             3924..3947
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3951..3974
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3978..4001
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4017..4073
                     /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             4086..4880
                     /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(4140..4159)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4750..4769
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5607..5623)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5616..5638)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(5842..6657)
                     /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     6565..6584
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6649..6668
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 aggcagacct ctcgacggag ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
       61 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
      121 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
      181 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
      241 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      301 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
      361 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
      421 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
      481 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      541 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      601 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      661 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
      721 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
      781 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagaagctc
      841 gcacattcag cagcgttttt cagcgcgttt tcgatcaacg tttcaatgtt ggtatcaaca
      901 ccaggtttaa ctttgaactt atcggcactg acggttactg attttgaact tttgctttgc
      961 cacggaacgg tctgcgttgt cgggaagatg cgtgatctga tccttcaact cagcaaaagt
     1021 tcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     1081 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     1141 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     1201 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcttgcat
     1261 gcctgcaggt cgactctaga ggatccccgg gtaccgagct cgaattagct tgcatgcctg
     1321 caggtcaatt ccctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     1381 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     1441 tgggcgtgga tagcggtttg actcacgggg atttcaagtc tccaccccat tgacgtcaat
     1501 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     1561 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
     1621 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccataggaga
     1681 caccgggacc gatccagcct ggggatcccg accatggctt cgtacccctg ccatcaacac
     1741 gcgtctgcgt tcgaccaggc tgcgcgttct cgcggccata gcaaccgacg tacggcgttg
     1801 cgccctcgcc ggcagcaaga agccacggaa gtccgcctgg agcagaaaat gcccacgcta
     1861 ctgcgggttt atatagacgg tcctcacggg atggggaaaa ccaccaccac gcaactgctg
     1921 gtggccctgg gttcgcgcga cgatatcgtc tacgtacccg agccgatgac ttactggcag
     1981 gtgctggggg cttccgagac aatcgcgaac atctacacca cacaacaccg cctcgaccag
     2041 ggtgagatat cggccgggga cgcggcggtg gtaatgacaa gcgcccagat aacaatgggc
     2101 atgccttatg ccgtgaccga cgccgttctg gctcctcata tcggggggga ggctgggagc
     2161 tcacatgccc cgcccccggc cctcaccctc atcttcgacc gccatcccat cgccgccctc
     2221 ctgtgctacc cggccgcgcg ataccttatg ggcagcatga ccccccaggc cgtgctggcg
     2281 ttcgtggccc tcatcccgcc gaccttgccc ggcacaaaca tcgtgttggg ggcccttccg
     2341 gaggacagac acatcgaccg cctggccaaa cgccagcgcc ccggcgagcg gcttgacctg
     2401 gctatgctgg ccgcgattcg ccgcgtttac gggctgcttg ccaatacggt gcggtatctg
     2461 cagggcggcg ggtcgtggcg ggaggattgg ggacagcttt cggggacggc cgtgccgccc
     2521 cagggtgccg agccccagag caacgcgggc ccacgacccc atatcgggga cacgttattt
     2581 accctgtttc gggcccccga gttgctggcc cccaacggcg acctgtacaa cgtgtttgcc
     2641 tgggccttgg acgtcttggc caaacgcctc cgtcccatgc acgtctttat cctggattac
     2701 gaccaatcgc ccgccggctg ccgggacgcc ctgctgcaac ttacctccgg gatggtccag
     2761 acccacgtca ccacccccgg ctccataccg acgatctgcg acctggcgcg cacgtttgcc
     2821 cgggagatgg gggaggctaa ctgaggatcc cacgtcacta ttgtatactc tatattatac
     2881 tctatgttat actctgtaat cctactcaat aaacgtgtca cgcctgtgaa accgtactaa
     2941 gtctcccgtg tattcttatc accatcaggt gacatcctcg cccaggctgt caatcatgcc
     3001 ggtatcgatt ccagtagcac cggccccacg ctgacaaccc actcttgcag cgttagcagc
     3061 gcccctctta acaagccgac ccccaccagc gtcgcggtta ctaacactcc tctccccgac
     3121 ctgcagccca gctagctact gccgcctcta aacccctcca gccactaggc tgtgtcccac
     3181 agtcagtgtc acccagtgaa caggcattac ccccacatct ggaaccagcc tggccccaag
     3241 ggctacggca taactcagta ccaggtagag ttggccccac agagtacctt tccccagata
     3301 tgcaacgcca gcgaaagacc aagcgcaaaa ccaaagagca gctggctatc cttaaatcct
     3361 tttttttaca gtgccagtgg gcacggcgtg aggattacca aaagttagaa cagatcactg
     3421 gtttacctcg gcctgagatc attcagtggt ttggtgacac acgctatgcc ttgaagcatg
     3481 ggcaactaaa gtggtttcgg gacaacgcag tacctggtgc ccctagtttc caagacccag
     3541 caattcctac accaccacca tcaacccgct ccttgaacga aagggctgag acaccacctc
     3601 tgccgatccc tccaccccca ccggatatac aacccttgga gaggtactgg gcagcccacc
     3661 aacagctacg ggaaactgat atccctcaat tgagtcaggc atcaaggctt agcacccagc
     3721 aggtgctgga ttggtttgac tctcgattac ctcagccagc tgaggtggta gtttgtctag
     3781 atgaagaaga ggaagaggag gaggaagaac tgccagaaga tgatgaggaa gaggaggagg
     3841 aggaggagga agatgatgat gatgatgatg atgtgatcat acaagacggg agcggaggag
     3901 gttccggtgg aggtggttct ggagattaca aggatgacga cgataagggc gattacaagg
     3961 atgacgacga taagggagat tacaaggatg acgacgataa ggtttcagga agcggagcta
     4021 ctaacttcag cctgctgaag caggctggag acgtggagga gaaccctgga cctggatcgt
     4081 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4141 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4201 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4261 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4321 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4381 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4441 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4501 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4561 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4621 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4681 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcc gaccgctatc
     4741 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4801 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4861 ttcttgacga gttcttctga ttcgaacatc gtggggagct gggggagggg aggtctgtta
     4921 ctgaaagatg aaccaactta gtaactgttt aaacaaagaa accacatttt taaaattacc
     4981 ttgtggcaga gaactaaaaa gctttgtgtt cttcagggtg ggtggcaggg ggatatagtg
     5041 agggtggacc agggagaatg accatagggc actaagtaag gctgggattg gatcagcaga
     5101 aatccaaccc tctaacctta gggtagggag tgctaaggat ctggggaaac catgggctgg
     5161 gaagctgctc ttgctctcct cgtgtctgct gtttttttcc cttggtgtac tatacagagg
     5221 ccagatgttg gcaccacctc tccaggagga ttggaaagga ggagtaaagg attctgattt
     5281 gattgatgat tccagtgcat ccccaatccc accatcttac ctggaatata aggctgcctt
     5341 gtaccccttt tctgagcaca agtctgtgcg taatgcaact gactctctta cttttttctt
     5401 agtaactgat catttcctag acaaccaaga ttctcaataa gtcccagtct catcacaaat
     5461 attaatattt ccttttcctc ataccaactt gactatgttt cacccgcttc atgactaaat
     5521 cacaggtggc agaatttttc agttcttttt ttttcttgga aagtcctctc cactgactgt
     5581 agcctccaat tcactggccg tcgttcactg gccgtcgttt tacaacgtcg tgactgggaa
     5641 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     5701 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     5761 tgcgatttat tcaacaaagc cgccgtcccg tcaagtcagc gtaatgctct gccagtgtta
     5821 caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaatat
     5881 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     5941 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     6001 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     6061 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat gcatttcttt
     6121 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     6181 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
     6241 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     6301 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     6361 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     6421 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     6481 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     6541 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     6601 catgttggaa tttaatcgcg gcttcgagca agacgtttcc cgttgaatat ggctcataac
     6661 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt
     6721 atcttgtgca atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg
     6781 aacttttgct gagttgaagg atcagatcac gcatcttccc gacaacgcag accgttccgt
     6841 ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac aacaaagctc tcatcaaccg
     6901 tggctccctc actttctggc tggatgatgg ggcgattcag gcctggtatg agtcagcaac
     6961 accttcttca cg
//
