LOCUS       sequence-171005-        7153 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human MBD1_iso1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7153)
  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 7153)
  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_MBD1_iso1
FEATURES             Location/Qualifiers
     source          1..7153
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      45..633
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     534..553
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     787..804
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1104..1125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1140..1145,1146..1163,1164..1170)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1178..1194
                     /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     1183..1205
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1202..1218
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1202..1218
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1395..1597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1548..1568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1594..1618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1694..2824
                     /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             3950..3973
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3977..4000
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4004..4027
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4043..4099
                     /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             4112..4906
                     /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(4166..4185)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4776..4795
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5768..5784)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5777..5799)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(6003..6818)
                     /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     6726..6745
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6810..6829
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
       61 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
      121 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
      181 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
      241 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
      301 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
      361 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
      421 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
      481 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
      541 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
      601 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
      661 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
      721 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
      781 gagcgcagcg agtcagtgag cgaggaagcg gaagaagctc gcacattcag cagcgttttt
      841 cagcgcgttt tcgatcaacg tttcaatgtt ggtatcaaca ccaggtttaa ctttgaactt
      901 atcggcactg acggttactg attttgaact tttgctttgc cacggaacgg tctgcgttgt
      961 cgggaagatg cgtgatctga tccttcaact cagcaaaagt tcccaatacg caaaccgcct
     1021 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     1081 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     1141 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     1201 acaggaaaca gctatgacca tgattacgcc aagcttgcat gcctgcaggt cgactctaga
     1261 ggatccccgg gtaccgagct cgaattagct tgcatgcctg caggtcaatt ccctggcatt
     1321 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     1381 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     1441 actcacgggg atttcaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     1501 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     1561 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc
     1621 ctggagacgc catccacgct gttttgacct ccataggaga caccgggacc gatccagcct
     1681 ggggatcccg accatggctt cgtacccctg ccatcaacac gcgtctgcgt tcgaccaggc
     1741 tgcgcgttct cgcggccata gcaaccgacg tacggcgttg cgccctcgcc ggcagcaaga
     1801 agccacggaa gtccgcctgg agcagaaaat gcccacgcta ctgcgggttt atatagacgg
     1861 tcctcacggg atggggaaaa ccaccaccac gcaactgctg gtggccctgg gttcgcgcga
     1921 cgatatcgtc tacgtacccg agccgatgac ttactggcag gtgctggggg cttccgagac
     1981 aatcgcgaac atctacacca cacaacaccg cctcgaccag ggtgagatat cggccgggga
     2041 cgcggcggtg gtaatgacaa gcgcccagat aacaatgggc atgccttatg ccgtgaccga
     2101 cgccgttctg gctcctcata tcggggggga ggctgggagc tcacatgccc cgcccccggc
     2161 cctcaccctc atcttcgacc gccatcccat cgccgccctc ctgtgctacc cggccgcgcg
     2221 ataccttatg ggcagcatga ccccccaggc cgtgctggcg ttcgtggccc tcatcccgcc
     2281 gaccttgccc ggcacaaaca tcgtgttggg ggcccttccg gaggacagac acatcgaccg
     2341 cctggccaaa cgccagcgcc ccggcgagcg gcttgacctg gctatgctgg ccgcgattcg
     2401 ccgcgtttac gggctgcttg ccaatacggt gcggtatctg cagggcggcg ggtcgtggcg
     2461 ggaggattgg ggacagcttt cggggacggc cgtgccgccc cagggtgccg agccccagag
     2521 caacgcgggc ccacgacccc atatcgggga cacgttattt accctgtttc gggcccccga
     2581 gttgctggcc cccaacggcg acctgtacaa cgtgtttgcc tgggccttgg acgtcttggc
     2641 caaacgcctc cgtcccatgc acgtctttat cctggattac gaccaatcgc ccgccggctg
     2701 ccgggacgcc ctgctgcaac ttacctccgg gatggtccag acccacgtca ccacccccgg
     2761 ctccataccg acgatctgcg acctggcgcg cacgtttgcc cgggagatgg gggaggctaa
     2821 ctgaggatcc cacgtcacta ttgtatactc tatattatac tctatgttat actctgtaat
     2881 cctactcaat aaacgtgtca cgcctgtgaa accgtactaa gtctcccgtg tattcttatc
     2941 accatcaggt gacatcctcg cccaggctgt caatcatgcc ggtatcgatt ccagtagcac
     3001 cggccccacg ctgacaaccc actcttgcag cgttagcagc gcccctctta acaagccgac
     3061 ccccaccagc gtcgcggtta ctaacactcc tctccccgac ctgcagccca gctgtctcca
     3121 ggagacctct gggtgggctg ttgcctgaca ttgagcaatg tgtaccagag aaatggtgct
     3181 gggcttgcct gggcagtggt atgggcagga gatctgggtt ccgagctggc tttgtgagct
     3241 gggttcacac gtagttgtgt gatctctggt tgccctgcac ttcctggagc ggcctctgtg
     3301 gaagggcttg ggctggatct cttgagcctt tcctgagcct attgaggtta ttaacttcag
     3361 atgccttgca gaactgtccc tgtagtcctg gcatagaaca aagggtagat gcctgtaagg
     3421 actagggtgg ttgggaggac agaggagtgc tggctcctgc agaatattga cagaccctgc
     3481 atagagcccc agacactggc gatgtaaact attggcctaa gcacttggat tcctttgctc
     3541 ctacaaacca gcccaccagg ctagaaaccc tttcatctat acttggccaa ttacagaagt
     3601 attgattgag ttcctgttgg gtgcacagca ttgaggacaa tgagccctgc aaagcttacc
     3661 cttcaggaga cacaggatac ctaccttcag ggagcttata atctgactat gtacaagctg
     3721 tctataatgg tagtttttag aagatataaa aaaagataaa ggataattaa ggggaagtgg
     3781 atggaggaca cagaatagta ggaaatggtt ttgtaatgct ccattgcgca catagcaaac
     3841 ctcattcaac atacagacct gctccctttt ccctaggtcc aaagacctta aaaaacctgg
     3901 agctagaaag caggggagcg gaggaggttc cggtggaggt ggttctggag attacaagga
     3961 tgacgacgat aagggcgatt acaaggatga cgacgataag ggagattaca aggatgacga
     4021 cgataaggtt tcaggaagcg gagctactaa cttcagcctg ctgaagcagg ctggagacgt
     4081 ggaggagaac cctggacctg gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4141 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4201 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4261 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     4321 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4381 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4441 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4501 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4561 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4621 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4681 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4741 ccggctgggt gtggccgacc gctatcagga catagcgttg gctacccgtg atattgctga
     4801 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4861 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgattcg aacatcggag
     4921 gcttctacag actgtaggat tcaaggtgat atttgcagac tggctttatg agagacaaca
     4981 ctgatctact aggggctgga ccctagattg gttgccaggg cttgtgtgtg aatcaaccct
     5041 aggaggaaaa acctactacc aaaccagaag agcaggccta agagtacttt gagcttctag
     5101 aagaagtaag cgggaaagga agagaaagag tagaaagttg gtgcatcttc tcctccccaa
     5161 gagcctgagt ataagaagtt ggattttaag ggtgggggag ctgggttgga gaaaggtaat
     5221 ataattaaag ccaccacaaa acaacctaaa caagccatgt tcatgagtaa ttgtaagtgg
     5281 cttctggatg agacctggca gtgtctctac atacttcctc agggagatag caattgaaaa
     5341 tgtaattgaa taaataagga aaggaactgc tatttggtgt gttcctctgt ggacccagtt
     5401 tatttaatcc ttaaaataat atttttggga agacattgat tttgtccatt tatgtggatg
     5461 ggaaaactga gactcattaa tttgtccaat gtcgaacagc tggtgttagt agagcctaga
     5521 ctcaaaccca gcttttaaag cccttgttct ttactccact aaactgtaac ctgtaatcct
     5581 gtatgcacta acaggagttt tcggtgacct tgaaaacaca gattactata attactataa
     5641 taaatccctt acattcctgt ctctagcatg gtttagggag tgccaaggct caatgtagat
     5701 ggggtcacct tgtcacttgg aaagtcctct ccactgactg tagcctccaa ttcactggcc
     5761 gtcgttcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     5821 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     5881 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgcgattta ttcaacaaag
     5941 ccgccgtccc gtcaagtcag cgtaatgctc tgccagtgtt acaaccaatt aaccaattct
     6001 gattagaaaa actcatcgag catcaaatga aactgcaata tattcatatc aggattatca
     6061 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     6121 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     6181 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     6241 actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc
     6301 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     6361 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     6421 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     6481 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     6541 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     6601 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     6661 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     6721 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     6781 ggcttcgagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     6841 atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat
     6901 cagagatttt gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag
     6961 gatcagatca cgcatcttcc cgacaacgca gaccgttccg tggcaaagca aaagttcaaa
     7021 atcaccaact ggtccaccta caacaaagct ctcatcaacc gtggctccct cactttctgg
     7081 ctggatgatg gggcgattca ggcctggtat gagtcagcaa caccttcttc acgaggcaga
     7141 cctctcgacg gag
//
