LOCUS       sequence-170780-        6970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human MIER3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6970)
  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 6970)
  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_MIER3
FEATURES             Location/Qualifiers
     source          1..6970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      55..643
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     544..563
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     797..814
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1114..1135
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1150..1155,1156..1173,1174..1180)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1188..1204
                     /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     1193..1215
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1212..1228
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1212..1228
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1405..1607
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1558..1578
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1604..1628
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1704..2834
                     /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             3868..3891
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3895..3918
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3922..3945
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3961..4017
                     /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             4030..4824
                     /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(4084..4103)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4694..4713
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5595..5611)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5604..5626)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(5830..6645)
                     /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     6553..6572
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6637..6656
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ctcgacggag ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
       61 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
      121 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
      181 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
      241 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
      301 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
      361 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
      421 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
      481 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
      541 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
      601 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
      661 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
      721 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
      781 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagaagctc gcacattcag
      841 cagcgttttt cagcgcgttt tcgatcaacg tttcaatgtt ggtatcaaca ccaggtttaa
      901 ctttgaactt atcggcactg acggttactg attttgaact tttgctttgc cacggaacgg
      961 tctgcgttgt cgggaagatg cgtgatctga tccttcaact cagcaaaagt tcccaatacg
     1021 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     1081 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     1141 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     1201 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcttgcat gcctgcaggt
     1261 cgactctaga ggatccccgg gtaccgagct cgaattagct tgcatgcctg caggtcaatt
     1321 ccctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1381 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     1441 tagcggtttg actcacgggg atttcaagtc tccaccccat tgacgtcaat gggagtttgt
     1501 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     1561 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc
     1621 gtcagatcgc ctggagacgc catccacgct gttttgacct ccataggaga caccgggacc
     1681 gatccagcct ggggatcccg accatggctt cgtacccctg ccatcaacac gcgtctgcgt
     1741 tcgaccaggc tgcgcgttct cgcggccata gcaaccgacg tacggcgttg cgccctcgcc
     1801 ggcagcaaga agccacggaa gtccgcctgg agcagaaaat gcccacgcta ctgcgggttt
     1861 atatagacgg tcctcacggg atggggaaaa ccaccaccac gcaactgctg gtggccctgg
     1921 gttcgcgcga cgatatcgtc tacgtacccg agccgatgac ttactggcag gtgctggggg
     1981 cttccgagac aatcgcgaac atctacacca cacaacaccg cctcgaccag ggtgagatat
     2041 cggccgggga cgcggcggtg gtaatgacaa gcgcccagat aacaatgggc atgccttatg
     2101 ccgtgaccga cgccgttctg gctcctcata tcggggggga ggctgggagc tcacatgccc
     2161 cgcccccggc cctcaccctc atcttcgacc gccatcccat cgccgccctc ctgtgctacc
     2221 cggccgcgcg ataccttatg ggcagcatga ccccccaggc cgtgctggcg ttcgtggccc
     2281 tcatcccgcc gaccttgccc ggcacaaaca tcgtgttggg ggcccttccg gaggacagac
     2341 acatcgaccg cctggccaaa cgccagcgcc ccggcgagcg gcttgacctg gctatgctgg
     2401 ccgcgattcg ccgcgtttac gggctgcttg ccaatacggt gcggtatctg cagggcggcg
     2461 ggtcgtggcg ggaggattgg ggacagcttt cggggacggc cgtgccgccc cagggtgccg
     2521 agccccagag caacgcgggc ccacgacccc atatcgggga cacgttattt accctgtttc
     2581 gggcccccga gttgctggcc cccaacggcg acctgtacaa cgtgtttgcc tgggccttgg
     2641 acgtcttggc caaacgcctc cgtcccatgc acgtctttat cctggattac gaccaatcgc
     2701 ccgccggctg ccgggacgcc ctgctgcaac ttacctccgg gatggtccag acccacgtca
     2761 ccacccccgg ctccataccg acgatctgcg acctggcgcg cacgtttgcc cgggagatgg
     2821 gggaggctaa ctgaggatcc cacgtcacta ttgtatactc tatattatac tctatgttat
     2881 actctgtaat cctactcaat aaacgtgtca cgcctgtgaa accgtactaa gtctcccgtg
     2941 tattcttatc accatcaggt gacatcctcg cccaggctgt caatcatgcc ggtatcgatt
     3001 ccagtagcac cggccccacg ctgacaaccc actcttgcag cgttagcagc gcccctctta
     3061 acaagccgac ccccaccagc gtcgcggtta ctaacactcc tctccccgac ctgcagccca
     3121 gctttcaggg actatatgga tcgtttagta gatgaaacag aagctttggg tgggacggta
     3181 aatgcttcag ccttaacttc taaccggcct gagcctattc ctgatcaaca gctaaacatt
     3241 ctcaactcct tcactgccag tgacttgaca ggtaaaatga agacctttct gcatttagta
     3301 cagtttgttg caaaattgtg tcctctgttt gatgtcatag gaccacttac ttaaacaatt
     3361 ttccattcct ttttagcttt gaccaacagt gtagcaaccg tctgcgaccc cacagatgtg
     3421 aattgtttgg atgatagctt tcctccactg ggcaacacac cccgtggaca agttaatcat
     3481 gtgcctgttg taacagaaga gttactcacc ctgcccagca atggggaaag tgattgtttt
     3541 aatttatttg agactggatt ttatcactcg gagctaaacc ctatgaacat gtgcagtgaa
     3601 gagtcagaga gaccagcaaa aagattgaaa atgggcattg ccgtccctga atcctttatg
     3661 aatgaagttt ctgtaaataa cctgggtgtg gactttgaaa atcacacaca tcacatcacc
     3721 agtgccaaaa tggctgtttc tgtggctgac tttggcagtc tctctgccaa cgagaccaat
     3781 ggtttcatca gtgcccatgc tctgcatcag cacgcggccc tacattcgga agggagcgga
     3841 ggaggttccg gtggaggtgg ttctggagat tacaaggatg acgacgataa gggcgattac
     3901 aaggatgacg acgataaggg agattacaag gatgacgacg ataaggtttc aggaagcgga
     3961 gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctgga
     4021 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4081 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4141 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4201 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4261 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4321 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4381 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4441 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4501 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4561 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4621 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggccgaccgc
     4681 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4741 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4801 cgccttcttg acgagttctt ctgattcgaa catcgtgagg atcccggaac tgcgtgtgca
     4861 gcaccagtaa acttgagggg agctatcagg tttgcatagt ctttcactgg aagtttgaac
     4921 ctttctcact atgacatcag tgatgtcagt atgtatagaa ctatatcttg atttatcaag
     4981 agtattttca tttttcaatc cataaatgtg gaaatgaact atgatcagca gagttgggaa
     5041 ctaagattga actctgtggt gcagctttaa gctctgcttc tctcttcctc atctcccaat
     5101 acctcttccc cactcccttc tttttctatt cttttgtgtt ccccactgcc ctcactccca
     5161 attttaaaac ctgtagacag aggccaccag ctcaaaacca gcacagatgt tctgaactga
     5221 atggtgtggc ctgtttttgt gtaaattcct tttgccgtaa tggatgcagt ggaataacaa
     5281 tgtttacagg taccgatccc gatccctgct caatgtagca tttttttggt tttattttct
     5341 taataaaagc aggggtaggt ttctttaaac tgcacaaaca tgcaaggatt ttttaaaaat
     5401 ggaaacttct ctcatgttat tcaactagag cacttcagtt tacaaaacag caagtccatc
     5461 tttatggaag ccagcacaag gaactgtgtg caaataatga agacgcttgc ttggatcctg
     5521 tttcaaaatt ctagaccagg gccttggaaa gtcctctcca ctgactgtag cctccaattc
     5581 actggccgtc gttcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     5641 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     5701 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg cgatttattc
     5761 aacaaagccg ccgtcccgtc aagtcagcgt aatgctctgc cagtgttaca accaattaac
     5821 caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatatat tcatatcagg
     5881 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     5941 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     6001 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     6061 agtgacgact gaatccggtg agaatggcaa aagcttatgc atttctttcc agacttgttc
     6121 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     6181 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     6241 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     6301 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     6361 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     6421 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     6481 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     6541 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     6601 taatcgcggc ttcgagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     6661 actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat cttgtgcaat
     6721 gtaacatcag agattttgag acacaacgtg gctttgttga ataaatcgaa cttttgctga
     6781 gttgaaggat cagatcacgc atcttcccga caacgcagac cgttccgtgg caaagcaaaa
     6841 gttcaaaatc accaactggt ccacctacaa caaagctctc atcaaccgtg gctccctcac
     6901 tttctggctg gatgatgggg cgattcaggc ctggtatgag tcagcaacac cttcttcacg
     6961 aggcagacct
//
