LOCUS       sequence-168161-        7154 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human MIXL1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7154)
  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 7154)
  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_MIXL1
FEATURES             Location/Qualifiers
     source          1..7154
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             837..860
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             864..887
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             891..914
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             930..986
                     /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             999..1793
                     /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(1053..1072)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1663..1682
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2656..2672)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2665..2687)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2891..3706)
                     /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     3614..3633
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3698..3717
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4086..4674
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4575..4594
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4828..4845
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5145..5166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5181..5186,5187..5204,5205..5211)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5219..5235
                     /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     5224..5246
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5243..5259
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5243..5259
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5436..5638
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5589..5609
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5635..5659
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5735..6865
                     /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 cccgtggtcg gtgtccaagc gcagttttaa gtagcagccc tcatttaaac actgggagag
       61 tggacttgtt ttctctgcac ccctggcggt catacagctg tcagtctcaa cagctggcat
      121 gagcaaaata cctccttgat tagtttgaca agggagtgca gaccttaaaa aacaggtttt
      181 cttcataatt taatgcagtt tttttctaag aattctaagt gcttccattt ttgtgacccc
      241 atttttcatg aaaatttctc taactaaaga gtagatagaa ctaggcatta ttccaatgtt
      301 tccatctggg aatggggagg caaggaaagc caggtgttct gaacttctaa tcgtcaaccc
      361 aagtgctttc tgaagacaag gctctggaga ggaagccagg ggatgttaaa ctggaactta
      421 ccagtatgaa ctgtgaacat aatgttagca taaaaaactc caaccaaaag cagcttttat
      481 tttctcccct cttccaggta tggttccaga acaggcgtgc caagtctcgg cgtcagagtg
      541 ggaaatcctt ccaacctttg gctaggccgg agattatcct caaccactgt gctcctggaa
      601 ctgaaacgaa atgtctgaag ccccagctgc ctcttgaggt agatgtgaac tgcctgcccg
      661 aaccaaacgg ggttggaggg ggcatctctg actctagctc ccaaggtcag aattttgaaa
      721 cctgttcccc tctctctgaa gacattggtt caaagctgga ctcatgggag gaacacatct
      781 tttctgcctt tggtaacttc gggagcggag gaggttccgg tggaggtggt tctggagatt
      841 acaaggatga cgacgataag ggcgattaca aggatgacga cgataaggga gattacaagg
      901 atgacgacga taaggtttca ggaagcggag ctactaactt cagcctgctg aagcaggctg
      961 gagacgtgga ggagaaccct ggacctggat cgtttcgcat gattgaacaa gatggattgc
     1021 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     1081 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     1141 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     1201 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     1261 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     1321 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     1381 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     1441 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     1501 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     1561 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     1621 actgtggccg gctgggtgtg gccgaccgct atcaggacat agcgttggct acccgtgata
     1681 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     1741 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgattcgaac
     1801 atcttcggga taagctctga ggagccatga ctgacagcct gggagagaca catcagcata
     1861 ctgtcctttc tgacttccat gctaaggaca tgtccttgtt aaccttgatg atggttttga
     1921 cagcacctct cacatttgaa ggtaccccgc cactttgtca atgacgtttt aagcccacac
     1981 tcccacccca gagttcccgc attcgttttt acctgtgttc tctccaagcc tgcacattcc
     2041 attggtctgc atccctatgc cttcttgcca ggcctgtttt tagtttttgg actggttgtt
     2101 cagaactcat tattttcttc acaagaatgc ctcagcttga ctcagtttcc ccttgtgttt
     2161 gacagctgcc attttctcct ggtccctcca aggcttataa tcttaaagtc actctacccc
     2221 gtctcttcaa ccctcatcct aggtttatta ctttttaaaa ttgggcctgt catcttcacg
     2281 ttcaatcata gctccaatga ctctgcatgc agattatttc gacagccccc ttgcctctag
     2341 cttctcaact acttaaaaaa aattaccctc tgtgggccag gtgcagtggc tcactcctgc
     2401 aatctcagca ctttgggagg ccgagtgggt ggatcacctg aagtcaggag ttcaagacca
     2461 gcctggccaa tatggcgaaa ccccatctct gctaaaaata taaaacttag ctgggcacgg
     2521 tgacgggagc ctgtagtccc agctactcag gaggctgagg caggagaatc acttgagcct
     2581 gggaggtgga ggttgcagtg atccttggaa agtcctctcc actgactgta gcctccaatt
     2641 cactggccgt cgttcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     2701 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     2761 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gcgatttatt
     2821 caacaaagcc gccgtcccgt caagtcagcg taatgctctg ccagtgttac aaccaattaa
     2881 ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     2941 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     3001 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     3061 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     3121 gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt
     3181 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     3241 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
     3301 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     3361 aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta
     3421 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     3481 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     3541 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
     3601 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     3661 ttaatcgcgg cttcgagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat
     3721 tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa
     3781 tgtaacatca gagattttga gacacaacgt ggctttgttg aataaatcga acttttgctg
     3841 agttgaagga tcagatcacg catcttcccg acaacgcaga ccgttccgtg gcaaagcaaa
     3901 agttcaaaat caccaactgg tccacctaca acaaagctct catcaaccgt ggctccctca
     3961 ctttctggct ggatgatggg gcgattcagg cctggtatga gtcagcaaca ccttcttcac
     4021 gaggcagacc tctcgacgga gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4081 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4141 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4201 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4261 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4321 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4381 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4441 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4501 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4561 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4621 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4681 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     4741 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     4801 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagaagct
     4861 cgcacattca gcagcgtttt tcagcgcgtt ttcgatcaac gtttcaatgt tggtatcaac
     4921 accaggttta actttgaact tatcggcact gacggttact gattttgaac ttttgctttg
     4981 ccacggaacg gtctgcgttg tcgggaagat gcgtgatctg atccttcaac tcagcaaaag
     5041 ttcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     5101 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     5161 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     5221 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgca
     5281 tgcctgcagg tcgactctag aggatccccg ggtaccgagc tcgaattagc ttgcatgcct
     5341 gcaggtcaat tccctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     5401 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     5461 atgggcgtgg atagcggttt gactcacggg gatttcaagt ctccacccca ttgacgtcaa
     5521 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     5581 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
     5641 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccataggag
     5701 acaccgggac cgatccagcc tggggatccc gaccatggct tcgtacccct gccatcaaca
     5761 cgcgtctgcg ttcgaccagg ctgcgcgttc tcgcggccat agcaaccgac gtacggcgtt
     5821 gcgccctcgc cggcagcaag aagccacgga agtccgcctg gagcagaaaa tgcccacgct
     5881 actgcgggtt tatatagacg gtcctcacgg gatggggaaa accaccacca cgcaactgct
     5941 ggtggccctg ggttcgcgcg acgatatcgt ctacgtaccc gagccgatga cttactggca
     6001 ggtgctgggg gcttccgaga caatcgcgaa catctacacc acacaacacc gcctcgacca
     6061 gggtgagata tcggccgggg acgcggcggt ggtaatgaca agcgcccaga taacaatggg
     6121 catgccttat gccgtgaccg acgccgttct ggctcctcat atcggggggg aggctgggag
     6181 ctcacatgcc ccgcccccgg ccctcaccct catcttcgac cgccatccca tcgccgccct
     6241 cctgtgctac ccggccgcgc gataccttat gggcagcatg accccccagg ccgtgctggc
     6301 gttcgtggcc ctcatcccgc cgaccttgcc cggcacaaac atcgtgttgg gggcccttcc
     6361 ggaggacaga cacatcgacc gcctggccaa acgccagcgc cccggcgagc ggcttgacct
     6421 ggctatgctg gccgcgattc gccgcgttta cgggctgctt gccaatacgg tgcggtatct
     6481 gcagggcggc gggtcgtggc gggaggattg gggacagctt tcggggacgg ccgtgccgcc
     6541 ccagggtgcc gagccccaga gcaacgcggg cccacgaccc catatcgggg acacgttatt
     6601 taccctgttt cgggcccccg agttgctggc ccccaacggc gacctgtaca acgtgtttgc
     6661 ctgggccttg gacgtcttgg ccaaacgcct ccgtcccatg cacgtcttta tcctggatta
     6721 cgaccaatcg cccgccggct gccgggacgc cctgctgcaa cttacctccg ggatggtcca
     6781 gacccacgtc accacccccg gctccatacc gacgatctgc gacctggcgc gcacgtttgc
     6841 ccgggagatg ggggaggcta actgaggatc ccacgtcact attgtatact ctatattata
     6901 ctctatgtta tactctgtaa tcctactcaa taaacgtgtc acgcctgtga aaccgtacta
     6961 agtctcccgt gtattcttat caccatcagg tgacatcctc gcccaggctg tcaatcatgc
     7021 cggtatcgat tccagtagca ccggccccac gctgacaacc cactcttgca gcgttagcag
     7081 cgcccctctt aacaagccga cccccaccag cgtcgcggtt actaacactc ctctccccga
     7141 cctgcagccc agct
//
