LOCUS       sequence-190320-        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             1594..1617
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1621..1644
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1648..1671
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1687..1743
                     /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             1756..2550
                     /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(1810..1829)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2420..2439
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3321..3337)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3330..3352)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(3556..4371)
                     /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     4279..4298
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4363..4382
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4751..5339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5240..5259
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5493..5510
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5810..5831
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5846..5851,5852..5869,5870..5876)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5884..5900
                     /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     5889..5911
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5908..5924
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5908..5924
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6101..6303
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6254..6274
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6300..6324
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(6400..6712,1..818)
                     /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 accgcctcga ccagggtgag atatcggccg gggacgcggc ggtggtaatg acaagcgccc
       61 agataacaat gggcatgcct tatgccgtga ccgacgccgt tctggctcct catatcgggg
      121 gggaggctgg gagctcacat gccccgcccc cggccctcac cctcatcttc gaccgccatc
      181 ccatcgccgc cctcctgtgc tacccggccg cgcgatacct tatgggcagc atgacccccc
      241 aggccgtgct ggcgttcgtg gccctcatcc cgccgacctt gcccggcaca aacatcgtgt
      301 tgggggccct tccggaggac agacacatcg accgcctggc caaacgccag cgccccggcg
      361 agcggcttga cctggctatg ctggccgcga ttcgccgcgt ttacgggctg cttgccaata
      421 cggtgcggta tctgcagggc ggcgggtcgt ggcgggagga ttggggacag ctttcgggga
      481 cggccgtgcc gccccagggt gccgagcccc agagcaacgc gggcccacga ccccatatcg
      541 gggacacgtt atttaccctg tttcgggccc ccgagttgct ggcccccaac ggcgacctgt
      601 acaacgtgtt tgcctgggcc ttggacgtct tggccaaacg cctccgtccc atgcacgtct
      661 ttatcctgga ttacgaccaa tcgcccgccg gctgccggga cgccctgctg caacttacct
      721 ccgggatggt ccagacccac gtcaccaccc ccggctccat accgacgatc tgcgacctgg
      781 cgcgcacgtt tgcccgggag atgggggagg ctaactgagg atcccacgtc actattgtat
      841 actctatatt atactctatg ttatactctg taatcctact caataaacgt gtcacgcctg
      901 tgaaaccgta ctaagtctcc cgtgtattct tatcaccatc aggtgacatc ctcgcccagg
      961 ctgtcaatca tgccggtatc gattccagta gcaccggccc cacgctgaca acccactctt
     1021 gcagcgttag cagcgcccct cttaacaagc cgacccccac cagcgtcgcg gttactaaca
     1081 ctcctctccc cgacctgcag cccagctttt ttgagacgga gtctcactct gtcaccaagg
     1141 ctggagtgca gtggtgtgat gtcagctcac tgcaacttcc acctcctggg ttcaggcgat
     1201 tctcatacct cagcctcctg tgtagctggg attacaggca tgtgccacca cacctaattt
     1261 ttgtattttt agtagagacg gggtttcact atgttggcca ggctggtctt gaaattcagg
     1321 tgatccaccc gccttggcct cccgaagtgc tgggattata ggcgtgagcc accacacctg
     1381 gcccccgggg ggctattttt aaagcacttt tctaaagtac atggctgatc taggcaggca
     1441 tcttggaaac tactctgcgt tgacttgggc attgtctatt tcttcctctg ctgccctcgc
     1501 cagaccctgc gggagattgt gattggcgtc ctgcaccaat tgaaaaacca gctttacggg
     1561 agcggaggag gttccggtgg aggtggttct ggagattaca aggatgacga cgataagggc
     1621 gattacaagg atgacgacga taagggagat tacaaggatg acgacgataa ggtttcagga
     1681 agcggagcta ctaacttcag cctgctgaag caggctggag acgtggagga gaaccctgga
     1741 cctggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     1801 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     1861 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     1921 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     1981 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     2041 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     2101 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     2161 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     2221 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     2281 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     2341 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcc
     2401 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     2461 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     2521 ttctatcgcc ttcttgacga gttcttctga ttcgaacatc cttggaaacc tggagaacag
     2581 ccaacaagag gcccttgaat ctctacgtgg ccactgaact gctgggcccg ggagactgga
     2641 ctacaacacc tcacactggt cagctggttt ctacttggtg tttggttttt cccagcccca
     2701 ttttatcttc agcggagccg cggtgtttgt tttgtgaaag cttctgatta atttattata
     2761 ttgacgataa aactcaaacc tacccagcct tccccccact ccatggaagt ccttgggatg
     2821 ggcgtctgct ctggacaccc caaagagctc ctgccctctc agccctttat tcaagcctca
     2881 gatttctgct catgatctac atagatttgg aaactgtttt cctctgtttt ggtctcttgg
     2941 gcaacatttt tggcccaagt ttgggcaaca tttggcccaa gtttgggcat tttggcagta
     3001 gctgtatggg agaaaaagag taagaggaaa tattcccaca gccatgaagg gtgaaagggc
     3061 accttgtgcc tagactaggg ctgcctggtc agtcccaggt gaggccaagg gctttctggc
     3121 catctcaggg aggggccacc aggttcctcc cctcacccca tattccatca ccttcctcct
     3181 ctgctctggg tggtaaggga agccctcccg gttcccacag gctatgatgc tgcatggcag
     3241 aggcaggtat aacacagcac tacatattct tggaaagtcc tctccactga ctgtagcctc
     3301 caattcactg gccgtcgttc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     3361 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     3421 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgcgat
     3481 ttattcaaca aagccgccgt cccgtcaagt cagcgtaatg ctctgccagt gttacaacca
     3541 attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca atatattcat
     3601 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     3661 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     3721 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     3781 accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac
     3841 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     3901 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     3961 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     4021 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     4081 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     4141 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     4201 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     4261 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     4321 ggaatttaat cgcggcttcg agcaagacgt ttcccgttga atatggctca taacacccct
     4381 tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg
     4441 tgcaatgtaa catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt
     4501 tgctgagttg aaggatcaga tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa
     4561 gcaaaagttc aaaatcacca actggtccac ctacaacaaa gctctcatca accgtggctc
     4621 cctcactttc tggctggatg atggggcgat tcaggcctgg tatgagtcag caacaccttc
     4681 ttcacgaggc agacctctcg acggagttcc actgagcgtc agaccccgta gaaaagatca
     4741 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4801 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4861 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4921 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4981 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5041 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5101 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5161 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5221 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5281 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5341 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5401 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     5461 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     5521 aagctcgcac attcagcagc gtttttcagc gcgttttcga tcaacgtttc aatgttggta
     5581 tcaacaccag gtttaacttt gaacttatcg gcactgacgg ttactgattt tgaacttttg
     5641 ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg atctgatcct tcaactcagc
     5701 aaaagttccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     5761 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     5821 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     5881 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     5941 ttgcatgcct gcaggtcgac tctagaggat ccccgggtac cgagctcgaa ttagcttgca
     6001 tgcctgcagg tcaattccct ggcattatgc ccagtacatg accttatggg actttcctac
     6061 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     6121 catcaatggg cgtggatagc ggtttgactc acggggattt caagtctcca ccccattgac
     6181 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     6241 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     6301 gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat
     6361 aggagacacc gggaccgatc cagcctgggg atcccgacca tggcttcgta cccctgccat
     6421 caacacgcgt ctgcgttcga ccaggctgcg cgttctcgcg gccatagcaa ccgacgtacg
     6481 gcgttgcgcc ctcgccggca gcaagaagcc acggaagtcc gcctggagca gaaaatgccc
     6541 acgctactgc gggtttatat agacggtcct cacgggatgg ggaaaaccac caccacgcaa
     6601 ctgctggtgg ccctgggttc gcgcgacgat atcgtctacg tacccgagcc gatgacttac
     6661 tggcaggtgc tgggggcttc cgagacaatc gcgaacatct acaccacaca ac
//
