LOCUS       sequence-245632-        6970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human SIX4_iso1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6970)
  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 6970)
  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_SIX4_iso1
FEATURES             Location/Qualifiers
     source          1..6970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             745..768
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             772..795
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             799..822
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             838..894
                     /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             907..1701
                     /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(961..980)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1571..1590
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2472..2488)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2481..2503)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2707..3522)
                     /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     3430..3449
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3514..3533
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3902..4490
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4391..4410
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4644..4661
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4961..4982
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4997..5002,5003..5020,5021..5027)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5035..5051
                     /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     5040..5062
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5059..5075
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5059..5075
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5252..5454
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5405..5425
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5451..5475
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5551..6681
                     /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 agctctcttg ctcccagtgc agtggtatac acggttccta atacaggcca gactatagga
       61 tctgtgaaac aggaaggctt ggaaaggagc ctggtatttt ctcagttgat gcctgtcaat
      121 cagaatgcac aagtaaatgc aaacctgtct tctgaaaaca tctcggggag tggcctgcat
      181 ccactggcct cctcattagt taatgtatct ccaactcaca atttttctct cagtccctct
      241 acactactaa atcccactga gctaaaccgc gacattgccg atagccaacc aatgtctgca
      301 ccggtggcaa gcaaatctac tgtgacatct gtcagcaaca ctaactatgc aactcttcag
      361 aactgctccc ttattactgg tcaagaccta ttgtcagtcc ctatgactca ggctgccctt
      421 ggggaaatag ttcctacagc tgaagatcag gtaggtcacc cctccccagc agtacatcag
      481 gattttgtcc aagaacatcg tttggttctg caatcggtag ctaacatgaa agagaatttc
      541 ttatcaaatt ctgagagcaa agcaacaagt agcttaatga tgctggactc taaatccaag
      601 tatgtcttag atggcatggt tgatactgtc tgtgaagacc tggaaacaga caaaaaagag
      661 cttgccaagc tccagactgt ccagctggat gaagatatgc aagacttagg gagcggagga
      721 ggttccggtg gaggtggttc tggagattac aaggatgacg acgataaggg cgattacaag
      781 gatgacgacg ataagggaga ttacaaggat gacgacgata aggtttcagg aagcggagct
      841 actaacttca gcctgctgaa gcaggctgga gacgtggagg agaaccctgg acctggatcg
      901 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
      961 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     1021 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     1081 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     1141 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     1201 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     1261 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     1321 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     1381 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     1441 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     1501 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc cgaccgctat
     1561 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     1621 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     1681 cttcttgacg agttcttctg attcgaacat cctctttttg gcatcagcgg caaatctttt
     1741 catgaagccc caaggacaca aaacattttc ccatttaaag gaaaacactc tagttttgca
     1801 agtatatgca tacaagagac tttagattga tctgcatgaa gatcacagtt aagtatacag
     1861 gagtagaact gcattattgc agcctttttg ttcacttata aatttctctt ttaaatagat
     1921 ggagacaaag gacaaggtga aatgtatcaa gtcaaagtga atcatttagt tgactctata
     1981 attctaaggt caaaatggaa cttgatagtt ttttaaatta aaaaatgtat acacctaaca
     2041 tagaaaatta aagatagctg cagaccatta gaaataatac aattgttttt gtttactttt
     2101 actccatggg cattgaaaag gttaagaaac ataaatggta catattttta aagttaagta
     2161 gcatgcatat atatatgcac acacacctct ttttcagcat tttttgagaa agtcttgggt
     2221 ctcaaacaca tttgtctcaa cacatttcca aatgtgattt ctaatagctc agtgtggctg
     2281 agaaagtgcg gaactccact aaaaactctg aaacttcagt ataaatgtaa atatatcagt
     2341 tctataagca ggatttgagt aattacagtg aattttattt ttaatgtata tgtttctctt
     2401 caggcgctat gaactgaaac ttggaaagtc ctctccactg actgtagcct ccaattcact
     2461 ggccgtcgtt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     2521 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     2581 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatgcga tttattcaac
     2641 aaagccgccg tcccgtcaag tcagcgtaat gctctgccag tgttacaacc aattaaccaa
     2701 ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
     2761 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     2821 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     2881 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     2941 gacgactgaa tccggtgaga atggcaaaag cttatgcatt tctttccaga cttgttcaac
     3001 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     3061 tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     3121 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     3181 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     3241 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     3301 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     3361 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     3421 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     3481 tcgcggcttc gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     3541 gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta
     3601 acatcagaga ttttgagaca caacgtggct ttgttgaata aatcgaactt ttgctgagtt
     3661 gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt tccgtggcaa agcaaaagtt
     3721 caaaatcacc aactggtcca cctacaacaa agctctcatc aaccgtggct ccctcacttt
     3781 ctggctggat gatggggcga ttcaggcctg gtatgagtca gcaacacctt cttcacgagg
     3841 cagacctctc gacggagttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3901 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3961 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4021 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4081 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4141 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4201 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4261 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4321 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4381 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4441 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4501 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4561 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4621 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gaagctcgca
     4681 cattcagcag cgtttttcag cgcgttttcg atcaacgttt caatgttggt atcaacacca
     4741 ggtttaactt tgaacttatc ggcactgacg gttactgatt ttgaactttt gctttgccac
     4801 ggaacggtct gcgttgtcgg gaagatgcgt gatctgatcc ttcaactcag caaaagttcc
     4861 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     4921 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     4981 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     5041 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cttgcatgcc
     5101 tgcaggtcga ctctagagga tccccgggta ccgagctcga attagcttgc atgcctgcag
     5161 gtcaattccc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     5221 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     5281 gcgtggatag cggtttgact cacggggatt tcaagtctcc accccattga cgtcaatggg
     5341 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     5401 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
     5461 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca taggagacac
     5521 cgggaccgat ccagcctggg gatcccgacc atggcttcgt acccctgcca tcaacacgcg
     5581 tctgcgttcg accaggctgc gcgttctcgc ggccatagca accgacgtac ggcgttgcgc
     5641 cctcgccggc agcaagaagc cacggaagtc cgcctggagc agaaaatgcc cacgctactg
     5701 cgggtttata tagacggtcc tcacgggatg gggaaaacca ccaccacgca actgctggtg
     5761 gccctgggtt cgcgcgacga tatcgtctac gtacccgagc cgatgactta ctggcaggtg
     5821 ctgggggctt ccgagacaat cgcgaacatc tacaccacac aacaccgcct cgaccagggt
     5881 gagatatcgg ccggggacgc ggcggtggta atgacaagcg cccagataac aatgggcatg
     5941 ccttatgccg tgaccgacgc cgttctggct cctcatatcg ggggggaggc tgggagctca
     6001 catgccccgc ccccggccct caccctcatc ttcgaccgcc atcccatcgc cgccctcctg
     6061 tgctacccgg ccgcgcgata ccttatgggc agcatgaccc cccaggccgt gctggcgttc
     6121 gtggccctca tcccgccgac cttgcccggc acaaacatcg tgttgggggc ccttccggag
     6181 gacagacaca tcgaccgcct ggccaaacgc cagcgccccg gcgagcggct tgacctggct
     6241 atgctggccg cgattcgccg cgtttacggg ctgcttgcca atacggtgcg gtatctgcag
     6301 ggcggcgggt cgtggcggga ggattgggga cagctttcgg ggacggccgt gccgccccag
     6361 ggtgccgagc cccagagcaa cgcgggccca cgaccccata tcggggacac gttatttacc
     6421 ctgtttcggg cccccgagtt gctggccccc aacggcgacc tgtacaacgt gtttgcctgg
     6481 gccttggacg tcttggccaa acgcctccgt cccatgcacg tctttatcct ggattacgac
     6541 caatcgcccg ccggctgccg ggacgccctg ctgcaactta cctccgggat ggtccagacc
     6601 cacgtcacca cccccggctc cataccgacg atctgcgacc tggcgcgcac gtttgcccgg
     6661 gagatggggg aggctaactg aggatcccac gtcactattg tatactctat attatactct
     6721 atgttatact ctgtaatcct actcaataaa cgtgtcacgc ctgtgaaacc gtactaagtc
     6781 tcccgtgtat tcttatcacc atcaggtgac atcctcgccc aggctgtcaa tcatgccggt
     6841 atcgattcca gtagcaccgg ccccacgctg acaacccact cttgcagcgt tagcagcgcc
     6901 cctcttaaca agccgacccc caccagcgtc gcggttacta acactcctct ccccgacctg
     6961 cagcccagct
//
