LOCUS       sequence-271134-        6969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human MLLT10_iso1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6969)
  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 6969)
  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_MLLT10_iso1
FEATURES             Location/Qualifiers
     source          1..6969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..203
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     154..174
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     200..224
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             300..1430
                     /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             2464..2487
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2491..2514
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2518..2541
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2557..2613
                     /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             2626..3420
                     /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(2680..2699)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3290..3309
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4190..4206)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4199..4221)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(4425..5240)
                     /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     5148..5167
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5232..5251
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5620..6208
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6109..6128
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6362..6379
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6679..6700
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6715..6720,6721..6738,6739..6745)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6753..6769
                     /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     6758..6780
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6777..6793
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6777..6793
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      181 gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
      241 cacgctgttt tgacctccat aggagacacc gggaccgatc cagcctgggg atcccgacca
      301 tggcttcgta cccctgccat caacacgcgt ctgcgttcga ccaggctgcg cgttctcgcg
      361 gccatagcaa ccgacgtacg gcgttgcgcc ctcgccggca gcaagaagcc acggaagtcc
      421 gcctggagca gaaaatgccc acgctactgc gggtttatat agacggtcct cacgggatgg
      481 ggaaaaccac caccacgcaa ctgctggtgg ccctgggttc gcgcgacgat atcgtctacg
      541 tacccgagcc gatgacttac tggcaggtgc tgggggcttc cgagacaatc gcgaacatct
      601 acaccacaca acaccgcctc gaccagggtg agatatcggc cggggacgcg gcggtggtaa
      661 tgacaagcgc ccagataaca atgggcatgc cttatgccgt gaccgacgcc gttctggctc
      721 ctcatatcgg gggggaggct gggagctcac atgccccgcc cccggccctc accctcatct
      781 tcgaccgcca tcccatcgcc gccctcctgt gctacccggc cgcgcgatac cttatgggca
      841 gcatgacccc ccaggccgtg ctggcgttcg tggccctcat cccgccgacc ttgcccggca
      901 caaacatcgt gttgggggcc cttccggagg acagacacat cgaccgcctg gccaaacgcc
      961 agcgccccgg cgagcggctt gacctggcta tgctggccgc gattcgccgc gtttacgggc
     1021 tgcttgccaa tacggtgcgg tatctgcagg gcggcgggtc gtggcgggag gattggggac
     1081 agctttcggg gacggccgtg ccgccccagg gtgccgagcc ccagagcaac gcgggcccac
     1141 gaccccatat cggggacacg ttatttaccc tgtttcgggc ccccgagttg ctggccccca
     1201 acggcgacct gtacaacgtg tttgcctggg ccttggacgt cttggccaaa cgcctccgtc
     1261 ccatgcacgt ctttatcctg gattacgacc aatcgcccgc cggctgccgg gacgccctgc
     1321 tgcaacttac ctccgggatg gtccagaccc acgtcaccac ccccggctcc ataccgacga
     1381 tctgcgacct ggcgcgcacg tttgcccggg agatggggga ggctaactga ggatcccacg
     1441 tcactattgt atactctata ttatactcta tgttatactc tgtaatccta ctcaataaac
     1501 gtgtcacgcc tgtgaaaccg tactaagtct cccgtgtatt cttatcacca tcaggtgaca
     1561 tcctcgccca ggctgtcaat catgccggta tcgattccag tagcaccggc cccacgctga
     1621 caacccactc ttgcagcgtt agcagcgccc ctcttaacaa gccgaccccc accagcgtcg
     1681 cggttactaa cactcctctc cccgacctgc agcccagctt tcccccccag tgtttcacca
     1741 agtgtttttc ttgtgttaag aaatgcacta attacaaact tttcctaggt tatacaatat
     1801 gaaatgtatt taatggaaaa tcacagtaat ttaatgagct aatagagtgc tcataataga
     1861 gaacacagtg ctgggaggtg ctgtttgctt agtaccctgc tatgtgccag gtgtccttgc
     1921 aagatacagt gacatttgtg atttcagtct ttaataatag gctgtaactc catttttttt
     1981 atagattagt aaaaggaaac gcaaagaaag cccaaggaca aatagcaaat gtggaatcac
     2041 agccgatgac tttcttagcc cataaaaatt tcattctcta cagtaccatg ttacttctgt
     2101 ggaaaagttg attttggaag atgaattctg cattcattct ttacaagttc tgtccctcag
     2161 atgggcccac ctctttggtc tttttatact gttgaattta atgtttttaa agtatttact
     2221 agtaaattca tgtctatata ttacgttgca agtagccttg ccaacttgca agaaaaaagg
     2281 gagtaacttt ctataccaca ttttatctca gtcacagttt caaattcgga gaatattatc
     2341 aaaagttgat ttagctaacg catctgctct tttgtttacc tgcagagact tagtgataaa
     2401 actgggcctg tagctcaaga gaaaagtggg agcggaggag gttccggtgg aggtggttct
     2461 ggagattaca aggatgacga cgataagggc gattacaagg atgacgacga taagggagat
     2521 tacaaggatg acgacgataa ggtttcagga agcggagcta ctaacttcag cctgctgaag
     2581 caggctggag acgtggagga gaaccctgga cctggatcgt ttcgcatgat tgaacaagat
     2641 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2701 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2761 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     2821 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     2881 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     2941 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3001 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3061 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3121 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     3181 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3241 ttcatcgact gtggccggct gggtgtggcc gaccgctatc aggacatagc gttggctacc
     3301 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3361 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3421 ttcgaacatc aaacatctag aaattgccta tcctgctgtt ctagcacttc atctggctgc
     3481 ctttgcagtc cttttactac agctatgaag aaacgcaaca agaaactcaa tgcacaacaa
     3541 aggattaatt gctgcaagga cattcttgta aggctttgat tagttttctt gttgctttgt
     3601 tgcactgaaa tggaattccc atgcccctac cccttacccc agttttttga acatggaaag
     3661 aaaatttaat aactttttaa agtgacataa tttacatgca atatgtttat caactcaaga
     3721 atttaatata gttggtacac aactagtttt gtttataaat tggagatgca aatagcaaaa
     3781 ctaaatactt gctccattta caaactactt gattttattg tacaagttga aatatgctct
     3841 tttgtttggg ttacagtatg cttgctctaa gtcaaattcc aaggaactaa tttcttctcc
     3901 tggagttgca ttgattcagt attacaaata tatagcacat cacctgggac ttggcaatct
     3961 ttgttaaaaa aaaatttcct ttctaatggg atttggccaa ttttggtaat gaagttagga
     4021 tggtaatgtc tgcatctgct aaaggtaatt ttcttttgag aattgctttc tttagtgtta
     4081 agacctactc atattttgaa gaaatcttga gttaagtgag ttctgaggct gctggggctt
     4141 ggaaagtcct ctccactgac tgtagcctcc aattcactgg ccgtcgttca ctggccgtcg
     4201 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     4261 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     4321 agttgcgcag cctgaatggc gaatgcgatt tattcaacaa agccgccgtc ccgtcaagtc
     4381 agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg
     4441 agcatcaaat gaaactgcaa tatattcata tcaggattat caataccata tttttgaaaa
     4501 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     4561 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     4621 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     4681 ggcaaaagct tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     4741 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     4801 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     4861 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     4921 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     4981 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     5041 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     5101 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     5161 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcttcga gcaagacgtt
     5221 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
     5281 attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca
     5341 acgtggcttt gttgaataaa tcgaactttt gctgagttga aggatcagat cacgcatctt
     5401 cccgacaacg cagaccgttc cgtggcaaag caaaagttca aaatcaccaa ctggtccacc
     5461 tacaacaaag ctctcatcaa ccgtggctcc ctcactttct ggctggatga tggggcgatt
     5521 caggcctggt atgagtcagc aacaccttct tcacgaggca gacctctcga cggagttcca
     5581 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5641 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5701 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5761 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5821 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5881 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5941 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6001 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6061 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6121 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6181 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6241 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6301 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6361 cagcgagtca gtgagcgagg aagcggaaga agctcgcaca ttcagcagcg tttttcagcg
     6421 cgttttcgat caacgtttca atgttggtat caacaccagg tttaactttg aacttatcgg
     6481 cactgacggt tactgatttt gaacttttgc tttgccacgg aacggtctgc gttgtcggga
     6541 agatgcgtga tctgatcctt caactcagca aaagttccca atacgcaaac cgcctctccc
     6601 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     6661 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     6721 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     6781 aaacagctat gaccatgatt acgccaagct tgcatgcctg caggtcgact ctagaggatc
     6841 cccgggtacc gagctcgaat tagcttgcat gcctgcaggt caattccctg gcattatgcc
     6901 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     6961 attaccatg
//
