LOCUS       sequence-271686-        6964 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human ELK1_iso1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6964)
  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 6964)
  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_ELK1_iso1
FEATURES             Location/Qualifiers
     source          1..6964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        121..323
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     274..294
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     320..344
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             420..1550
                     /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             2584..2607
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2611..2634
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2638..2661
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2677..2733
                     /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             2746..3540
                     /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(2800..2819)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3410..3429
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4305..4321)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4314..4336)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(4540..5355)
                     /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     5263..5282
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5347..5366
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5735..6323
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6224..6243
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6477..6494
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6794..6815
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6830..6835,6836..6853,6854..6860)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6868..6884
                     /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     6873..6895
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6892..6908
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6892..6908
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cgagctcgaa ttagcttgca tgcctgcagg tcaattccct ggcattatgc ccagtacatg
       61 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      121 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      181 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      241 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      301 gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
      361 cacgctgttt tgacctccat aggagacacc gggaccgatc cagcctgggg atcccgacca
      421 tggcttcgta cccctgccat caacacgcgt ctgcgttcga ccaggctgcg cgttctcgcg
      481 gccatagcaa ccgacgtacg gcgttgcgcc ctcgccggca gcaagaagcc acggaagtcc
      541 gcctggagca gaaaatgccc acgctactgc gggtttatat agacggtcct cacgggatgg
      601 ggaaaaccac caccacgcaa ctgctggtgg ccctgggttc gcgcgacgat atcgtctacg
      661 tacccgagcc gatgacttac tggcaggtgc tgggggcttc cgagacaatc gcgaacatct
      721 acaccacaca acaccgcctc gaccagggtg agatatcggc cggggacgcg gcggtggtaa
      781 tgacaagcgc ccagataaca atgggcatgc cttatgccgt gaccgacgcc gttctggctc
      841 ctcatatcgg gggggaggct gggagctcac atgccccgcc cccggccctc accctcatct
      901 tcgaccgcca tcccatcgcc gccctcctgt gctacccggc cgcgcgatac cttatgggca
      961 gcatgacccc ccaggccgtg ctggcgttcg tggccctcat cccgccgacc ttgcccggca
     1021 caaacatcgt gttgggggcc cttccggagg acagacacat cgaccgcctg gccaaacgcc
     1081 agcgccccgg cgagcggctt gacctggcta tgctggccgc gattcgccgc gtttacgggc
     1141 tgcttgccaa tacggtgcgg tatctgcagg gcggcgggtc gtggcgggag gattggggac
     1201 agctttcggg gacggccgtg ccgccccagg gtgccgagcc ccagagcaac gcgggcccac
     1261 gaccccatat cggggacacg ttatttaccc tgtttcgggc ccccgagttg ctggccccca
     1321 acggcgacct gtacaacgtg tttgcctggg ccttggacgt cttggccaaa cgcctccgtc
     1381 ccatgcacgt ctttatcctg gattacgacc aatcgcccgc cggctgccgg gacgccctgc
     1441 tgcaacttac ctccgggatg gtccagaccc acgtcaccac ccccggctcc ataccgacga
     1501 tctgcgacct ggcgcgcacg tttgcccggg agatggggga ggctaactga ggatcccacg
     1561 tcactattgt atactctata ttatactcta tgttatactc tgtaatccta ctcaataaac
     1621 gtgtcacgcc tgtgaaaccg tactaagtct cccgtgtatt cttatcacca tcaggtgaca
     1681 tcctcgccca ggctgtcaat catgccggta tcgattccag tagcaccggc cccacgctga
     1741 caacccactc ttgcagcgtt agcagcgccc ctcttaacaa gccgaccccc accagcgtcg
     1801 cggttactaa cactcctctc cccgacctgc agcccagctg ggtctttctg tgggactgtt
     1861 tctaggttgg ggtgtggctt aaggaacctg agcagggctt tttggtctgg gatgtgcata
     1921 tggggtttca tgggtgggac atgagttgga agttgaccct gagctgttgg gatggtgact
     1981 tgagaggttt gagagcagca tttggatagg gtcttgtggg tggtcttgac caatgaggtc
     2041 tcagcaactg agggtgggct ctcgggagtg gggagtgggg ggtgtctgca gagtttgaga
     2101 gatcataggc atggcctcca tggttaccat ctaggtgtgt gcactttggg gagtgaccct
     2161 gggagtgtga gtgggaaatc tgtggtgggg ccgggaccaa gtcccttcca gcatcccaca
     2221 tctactttct actcacttga gctccgacca ccctctcccc acagaccccg gtgctgctga
     2281 cacccagctc gctgcctcct agcattcact tctggagcac cctgagtccc attgcgcccc
     2341 gtagcccggc caagctctcc ttccaggtag gatcctcttt ccccgttttg gacagggtga
     2401 gagacccaag aatggccccc atctgtgacc tctccctctt tttgccccca gtttccatcc
     2461 agtggcagcg cccaggtgca catcccttct atcagcgtgg atggcctctc gacccccgtg
     2521 gtgctctccc cagggcccca gaaacctggg agcggaggag gttccggtgg aggtggttct
     2581 ggagattaca aggatgacga cgataagggc gattacaagg atgacgacga taagggagat
     2641 tacaaggatg acgacgataa ggtttcagga agcggagcta ctaacttcag cctgctgaag
     2701 caggctggag acgtggagga gaaccctgga cctggatcgt ttcgcatgat tgaacaagat
     2761 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2821 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2881 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     2941 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3001 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3061 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3121 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3181 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3241 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     3301 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3361 ttcatcgact gtggccggct gggtgtggcc gaccgctatc aggacatagc gttggctacc
     3421 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3481 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3541 ttcgaacatc ccaccaccac caccacccct tctggggtca ctccatccat gctctctcca
     3601 gccagccatc tcaaggagaa acatagttca actgaaagac tcatgctctg attgtggtgg
     3661 ggtggggatc cttgggaaga attactccca agagtaactc tcattatctc ctccacagaa
     3721 aacacacagc ttccacaact tctctgtttt ctgtcagtcc cccagtggcc gcccttacac
     3781 gtctcctact tcaatggtag gggcggttta tttatttatt ttttgaaggc cactgggagg
     3841 agcctgacct aaccttttag ggtggttagg acatctcccc cacctcccca cttttttccc
     3901 caagacaaga caatcgaggt ctggcttgag aacgaccttt ctttctttat ttctcagcct
     3961 gcccttgggg agatgaggga gccctgtctg cgtttttgga tgtgagtaga agagttagtt
     4021 tgttttgttt tattattcct ggccatactc aggggtccag gaagaatttg taccatttaa
     4081 tgggttggga gtcttggcca aggaagaatc acacccttgg aatagaaatt tccacctccc
     4141 caacctttct ctcagacagc ttatcctttt caaccaactt tttggccagg gaggaatgtc
     4201 ccttttgttc ttccccctga gaagccattc ctttgtctgc caacctccct ggcttggaaa
     4261 gtcctctcca ctgactgtag cctccaattc actggccgtc gttcactggc cgtcgtttta
     4321 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     4381 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     4441 cgcagcctga atggcgaatg cgatttattc aacaaagccg ccgtcccgtc aagtcagcgt
     4501 aatgctctgc cagtgttaca accaattaac caattctgat tagaaaaact catcgagcat
     4561 caaatgaaac tgcaatatat tcatatcagg attatcaata ccatattttt gaaaaagccg
     4621 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
     4681 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
     4741 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
     4801 aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     4861 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac
     4921 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
     4981 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
     5041 tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
     5101 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
     5161 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     5221 cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     5281 cccatataaa tcagcatcca tgttggaatt taatcgcggc ttcgagcaag acgtttcccg
     5341 ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
     5401 tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg
     5461 gctttgttga ataaatcgaa cttttgctga gttgaaggat cagatcacgc atcttcccga
     5521 caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc accaactggt ccacctacaa
     5581 caaagctctc atcaaccgtg gctccctcac tttctggctg gatgatgggg cgattcaggc
     5641 ctggtatgag tcagcaacac cttcttcacg aggcagacct ctcgacggag ttccactgag
     5701 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5761 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5821 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5881 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5941 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6001 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6061 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6121 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6181 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6241 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6301 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6361 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6421 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     6481 agtcagtgag cgaggaagcg gaagaagctc gcacattcag cagcgttttt cagcgcgttt
     6541 tcgatcaacg tttcaatgtt ggtatcaaca ccaggtttaa ctttgaactt atcggcactg
     6601 acggttactg attttgaact tttgctttgc cacggaacgg tctgcgttgt cgggaagatg
     6661 cgtgatctga tccttcaact cagcaaaagt tcccaatacg caaaccgcct ctccccgcgc
     6721 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     6781 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     6841 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     6901 gctatgacca tgattacgcc aagcttgcat gcctgcaggt cgactctaga ggatccccgg
     6961 gtac
//
