LOCUS       sequence-182455-        6712 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human MLX.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6712)
  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 6712)
  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_MLX
FEATURES             Location/Qualifiers
     source          1..6712
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             487..510
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             514..537
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             541..564
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             580..636
                     /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             649..1443
                     /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(703..722)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1313..1332
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2214..2230)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2223..2245)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2449..3264)
                     /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
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3172..3191
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3256..3275
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3644..4232
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4133..4152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4386..4403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4703..4724
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4739..4744,4745..4762,4763..4769)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4777..4793
                     /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     4782..4804
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4801..4817
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4801..4817
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4994..5196
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5147..5167
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5193..5217
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5293..6423
                     /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 tttttgagac ggagtctcac tctgtcacca aggctggagt gcagtggtgt gatgtcagct
       61 cactgcaact tccacctcct gggttcaggc gattctcata cctcagcctc ctgtgtagct
      121 gggattacag gcatgtgcca ccacacctaa tttttgtatt tttagtagag acggggtttc
      181 actatgttgg ccaggctggt cttgaaattc aggtgatcca cccgccttgg cctcccgaag
      241 tgctgggatt ataggcgtga gccaccacac ctggcccccg gggggctatt tttaaagcac
      301 ttttctaaag tacatggctg atctaggcag gcatcttgga aactactctg cgttgacttg
      361 ggcattgtct atttcttcct ctgctgccct cgccagaccc tgcgggagat tgtgattggc
      421 gtcctgcacc aattgaaaaa ccagctttac gggagcggag gaggttccgg tggaggtggt
      481 tctggagatt acaaggatga cgacgataag ggcgattaca aggatgacga cgataaggga
      541 gattacaagg atgacgacga taaggtttca ggaagcggag ctactaactt cagcctgctg
      601 aagcaggctg gagacgtgga ggagaaccct ggacctggat cgtttcgcat gattgaacaa
      661 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
      721 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
      781 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
      841 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
      901 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
      961 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     1021 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     1081 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     1141 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     1201 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     1261 ggattcatcg actgtggccg gctgggtgtg gccgaccgct atcaggacat agcgttggct
     1321 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     1381 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     1441 tgattcgaac atccttggaa acctggagaa cagccaacaa gaggcccttg aatctctacg
     1501 tggccactga actgctgggc ccgggagact ggactacaac acctcacact ggtcagctgg
     1561 tttctacttg gtgtttggtt tttcccagcc ccattttatc ttcagcggag ccgcggtgtt
     1621 tgttttgtga aagcttctga ttaatttatt atattgacga taaaactcaa acctacccag
     1681 ccttcccccc actccatgga agtccttggg atgggcgtct gctctggaca ccccaaagag
     1741 ctcctgccct ctcagccctt tattcaagcc tcagatttct gctcatgatc tacatagatt
     1801 tggaaactgt tttcctctgt tttggtctct tgggcaacat ttttggccca agtttgggca
     1861 acatttggcc caagtttggg cattttggca gtagctgtat gggagaaaaa gagtaagagg
     1921 aaatattccc acagccatga agggtgaaag ggcaccttgt gcctagacta gggctgcctg
     1981 gtcagtccca ggtgaggcca agggctttct ggccatctca gggaggggcc accaggttcc
     2041 tcccctcacc ccatattcca tcaccttcct cctctgctct gggtggtaag ggaagccctc
     2101 ccggttccca caggctatga tgctgcatgg cagaggcagg tataacacag cactacatat
     2161 tcttggaaag tcctctccac tgactgtagc ctccaattca ctggccgtcg ttcactggcc
     2221 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     2281 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     2341 caacagttgc gcagcctgaa tggcgaatgc gatttattca acaaagccgc cgtcccgtca
     2401 agtcagcgta atgctctgcc agtgttacaa ccaattaacc aattctgatt agaaaaactc
     2461 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
     2521 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
     2581 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
     2641 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
     2701 gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc
     2761 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
     2821 acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
     2881 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
     2941 ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
     3001 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     3061 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     3121 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     3181 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggct tcgagcaaga
     3241 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag
     3301 ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga
     3361 cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc agatcacgca
     3421 tcttcccgac aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc
     3481 cacctacaac aaagctctca tcaaccgtgg ctccctcact ttctggctgg atgatggggc
     3541 gattcaggcc tggtatgagt cagcaacacc ttcttcacga ggcagacctc tcgacggagt
     3601 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3661 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3721 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3781 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3841 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3901 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3961 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4021 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4081 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4141 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4201 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4261 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4321 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4381 agcgcagcga gtcagtgagc gaggaagcgg aagaagctcg cacattcagc agcgtttttc
     4441 agcgcgtttt cgatcaacgt ttcaatgttg gtatcaacac caggtttaac tttgaactta
     4501 tcggcactga cggttactga ttttgaactt ttgctttgcc acggaacggt ctgcgttgtc
     4561 gggaagatgc gtgatctgat ccttcaactc agcaaaagtt cccaatacgc aaaccgcctc
     4621 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     4681 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     4741 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     4801 caggaaacag ctatgaccat gattacgcca agcttgcatg cctgcaggtc gactctagag
     4861 gatccccggg taccgagctc gaattagctt gcatgcctgc aggtcaattc cctggcatta
     4921 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     4981 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     5041 ctcacgggga tttcaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     5101 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     5161 aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc
     5221 tggagacgcc atccacgctg ttttgacctc cataggagac accgggaccg atccagcctg
     5281 gggatcccga ccatggcttc gtacccctgc catcaacacg cgtctgcgtt cgaccaggct
     5341 gcgcgttctc gcggccatag caaccgacgt acggcgttgc gccctcgccg gcagcaagaa
     5401 gccacggaag tccgcctgga gcagaaaatg cccacgctac tgcgggttta tatagacggt
     5461 cctcacggga tggggaaaac caccaccacg caactgctgg tggccctggg ttcgcgcgac
     5521 gatatcgtct acgtacccga gccgatgact tactggcagg tgctgggggc ttccgagaca
     5581 atcgcgaaca tctacaccac acaacaccgc ctcgaccagg gtgagatatc ggccggggac
     5641 gcggcggtgg taatgacaag cgcccagata acaatgggca tgccttatgc cgtgaccgac
     5701 gccgttctgg ctcctcatat cgggggggag gctgggagct cacatgcccc gcccccggcc
     5761 ctcaccctca tcttcgaccg ccatcccatc gccgccctcc tgtgctaccc ggccgcgcga
     5821 taccttatgg gcagcatgac cccccaggcc gtgctggcgt tcgtggccct catcccgccg
     5881 accttgcccg gcacaaacat cgtgttgggg gcccttccgg aggacagaca catcgaccgc
     5941 ctggccaaac gccagcgccc cggcgagcgg cttgacctgg ctatgctggc cgcgattcgc
     6001 cgcgtttacg ggctgcttgc caatacggtg cggtatctgc agggcggcgg gtcgtggcgg
     6061 gaggattggg gacagctttc ggggacggcc gtgccgcccc agggtgccga gccccagagc
     6121 aacgcgggcc cacgacccca tatcggggac acgttattta ccctgtttcg ggcccccgag
     6181 ttgctggccc ccaacggcga cctgtacaac gtgtttgcct gggccttgga cgtcttggcc
     6241 aaacgcctcc gtcccatgca cgtctttatc ctggattacg accaatcgcc cgccggctgc
     6301 cgggacgccc tgctgcaact tacctccggg atggtccaga cccacgtcac cacccccggc
     6361 tccataccga cgatctgcga cctggcgcgc acgtttgccc gggagatggg ggaggctaac
     6421 tgaggatccc acgtcactat tgtatactct atattatact ctatgttata ctctgtaatc
     6481 ctactcaata aacgtgtcac gcctgtgaaa ccgtactaag tctcccgtgt attcttatca
     6541 ccatcaggtg acatcctcgc ccaggctgtc aatcatgccg gtatcgattc cagtagcacc
     6601 ggccccacgc tgacaaccca ctcttgcagc gttagcagcg cccctcttaa caagccgacc
     6661 cccaccagcg tcgcggttac taacactcct ctccccgacc tgcagcccag ct
//
