LOCUS       sequence-196929-        6855 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human GATAD1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6855)
  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 6855)
  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_GATAD1
FEATURES             Location/Qualifiers
     source          1..6855
                     /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(2357..2373)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2366..2388)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2592..3407)
                     /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     3315..3334
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3399..3418
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3787..4375
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4276..4295
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4529..4546
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4846..4867
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4882..4887,4888..4905,4906..4912)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4920..4936
                     /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     4925..4947
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4944..4960
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4944..4960
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5137..5339
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5290..5310
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5336..5360
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5436..6566
                     /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 ccttcgggat tcctctactg cctcctcttt ctaatcccca gattctgata tgcatccttg
       61 tcctacagcg aggcagcatg gcatgaggtc agaacaccag ttctggagcc agactgtcta
      121 ggttcacagc ctgccattta ccggccatgt gactttggca agtttcttag tctctcttgc
      181 ctcactttcc tcatatgtaa aatgggaata ataatagtgc ctacctcaga aggttgatgt
      241 gaggaatgaa ggtattgata catgtaaact tagagcagtg tgggtacaaa ataaacatga
      301 tgcaagtgtt caatcactgt ttttgggaga atgccatatt ctttaagccg ttaaagaaga
      361 aaaaatgatt aagaataatt tcaaagtaat gcatgtttca agggctaatg ccaggttgct
      421 cccagagtgg tctctcccag tgtctagaaa ttttaacatc ttatgaaaat gatatatatg
      481 gtcaaaaatg tatttaacct ttcccttggc tgccttccag ggccagagga agatcttcca
      541 aggaagatgg aatacttgga atttgtttgt catgcacctt ctgagtattt caagtcacgg
      601 tcatcaccat ttcccacagt tcccaccaga ccagagaagg gctacatatg gactcatgtt
      661 gggcctactc ctgcaataac aattaaggaa tcagttgcca accacctagg 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 cactgggttt ccaggcctgg tgtggtggct
     1741 cacgcctgta gccccagcta ttgcaccact gctctccaag ctgggcaatg gagtcagatt
     1801 ctctttctta aaaaaccaca aaaaaactgg atttccagtt ctctaatatt cttagtacca
     1861 caagatatgt cataggtatc tttaaatgaa attcttagct ggaaaagtga ctaaaaagtt
     1921 tttctcctgc tacctagtaa taaacaaatc attgtttatt actggtcact tagaaaatta
     1981 aaagggatag ggccaggcac agtggcttat gcctgtaatt gcagcacttt tagaggccga
     2041 ggcaggcgga tcacctgagg tcgggaagtg gatcgcctga ggtcaggagt tcgagaccag
     2101 cctggccaac atggcgaaac cccgtcgcta ctaaaaatac aaaaattagc caggtgtggt
     2161 ggcatgtgcc tgtaatccca gctatttggg aggctgaggc aggagaatcg cctaaaccca
     2221 ggaggtggag gttgtagtga gccaagattg caccgctgtg ctccagcctg ggcaacagag
     2281 tgagactctt gtctcggaaa aaaacttgga aagtcctctc cactgactgt agcctccaat
     2341 tcactggccg tcgttcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     2401 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     2461 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgcgatttat
     2521 tcaacaaagc cgccgtcccg tcaagtcagc gtaatgctct gccagtgtta caaccaatta
     2581 accaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     2641 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     2701 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     2761 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     2821 tgagtgacga ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt
     2881 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     2941 attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa
     3001 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     3061 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt
     3121 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     3181 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     3241 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
     3301 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     3361 tttaatcgcg gcttcgagca agacgtttcc cgttgaatat ggctcataac accccttgta
     3421 ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca
     3481 atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct
     3541 gagttgaagg atcagatcac gcatcttccc gacaacgcag accgttccgt ggcaaagcaa
     3601 aagttcaaaa tcaccaactg gtccacctac aacaaagctc tcatcaaccg tggctccctc
     3661 actttctggc tggatgatgg ggcgattcag gcctggtatg agtcagcaac accttcttca
     3721 cgaggcagac ctctcgacgg agttccactg agcgtcagac cccgtagaaa agatcaaagg
     3781 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3841 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3901 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3961 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4021 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4081 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4141 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4201 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4261 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4321 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4381 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4441 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4501 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagaagc
     4561 tcgcacattc agcagcgttt ttcagcgcgt tttcgatcaa cgtttcaatg ttggtatcaa
     4621 caccaggttt aactttgaac ttatcggcac tgacggttac tgattttgaa cttttgcttt
     4681 gccacggaac ggtctgcgtt gtcgggaaga tgcgtgatct gatccttcaa ctcagcaaaa
     4741 gttcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     4801 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     4861 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     4921 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
     4981 atgcctgcag gtcgactcta gaggatcccc gggtaccgag ctcgaattag cttgcatgcc
     5041 tgcaggtcaa ttccctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     5101 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     5161 aatgggcgtg gatagcggtt tgactcacgg ggatttcaag tctccacccc attgacgtca
     5221 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     5281 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     5341 gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagga
     5401 gacaccggga ccgatccagc ctggggatcc cgaccatggc ttcgtacccc tgccatcaac
     5461 acgcgtctgc gttcgaccag gctgcgcgtt ctcgcggcca tagcaaccga cgtacggcgt
     5521 tgcgccctcg ccggcagcaa gaagccacgg aagtccgcct ggagcagaaa atgcccacgc
     5581 tactgcgggt ttatatagac ggtcctcacg ggatggggaa aaccaccacc acgcaactgc
     5641 tggtggccct gggttcgcgc gacgatatcg tctacgtacc cgagccgatg acttactggc
     5701 aggtgctggg ggcttccgag acaatcgcga acatctacac cacacaacac cgcctcgacc
     5761 agggtgagat atcggccggg gacgcggcgg tggtaatgac aagcgcccag ataacaatgg
     5821 gcatgcctta tgccgtgacc gacgccgttc tggctcctca tatcgggggg gaggctggga
     5881 gctcacatgc cccgcccccg gccctcaccc tcatcttcga ccgccatccc atcgccgccc
     5941 tcctgtgcta cccggccgcg cgatacctta tgggcagcat gaccccccag gccgtgctgg
     6001 cgttcgtggc cctcatcccg ccgaccttgc ccggcacaaa catcgtgttg ggggcccttc
     6061 cggaggacag acacatcgac cgcctggcca aacgccagcg ccccggcgag cggcttgacc
     6121 tggctatgct ggccgcgatt cgccgcgttt acgggctgct tgccaatacg gtgcggtatc
     6181 tgcagggcgg cgggtcgtgg cgggaggatt ggggacagct ttcggggacg gccgtgccgc
     6241 cccagggtgc cgagccccag agcaacgcgg gcccacgacc ccatatcggg gacacgttat
     6301 ttaccctgtt tcgggccccc gagttgctgg cccccaacgg cgacctgtac aacgtgtttg
     6361 cctgggcctt ggacgtcttg gccaaacgcc tccgtcccat gcacgtcttt atcctggatt
     6421 acgaccaatc gcccgccggc tgccgggacg ccctgctgca acttacctcc gggatggtcc
     6481 agacccacgt caccaccccc ggctccatac cgacgatctg cgacctggcg cgcacgtttg
     6541 cccgggagat gggggaggct aactgaggat cccacgtcac tattgtatac tctatattat
     6601 actctatgtt atactctgta atcctactca ataaacgtgt cacgcctgtg aaaccgtact
     6661 aagtctcccg tgtattctta tcaccatcag gtgacatcct cgcccaggct gtcaatcatg
     6721 ccggtatcga ttccagtagc accggcccca cgctgacaac ccactcttgc agcgttagca
     6781 gcgcccctct taacaagccg acccccacca gcgtcgcggt tactaacact cctctccccg
     6841 acctgcagcc cagct
//
