LOCUS       sequence-211492-        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"
     protein_bind    104..125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(140..145,146..163,164..170)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    178..194
                     /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     183..205
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     202..218
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     202..218
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        395..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             694..1824
                     /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             2858..2881
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2885..2908
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2912..2935
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2951..3007
                     /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             3020..3814
                     /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(3074..3093)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3684..3703
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4470..4486)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4479..4501)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(4705..5520)
                     /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     5428..5447
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5512..5531
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5900..6488
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6389..6408
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6642..6659
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
       61 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
      121 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
      181 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcttgcat
      241 gcctgcaggt cgactctaga ggatccccgg gtaccgagct cgaattagct tgcatgcctg
      301 caggtcaatt ccctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      361 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      421 tgggcgtgga tagcggtttg actcacgggg atttcaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccataggaga
      661 caccgggacc gatccagcct ggggatcccg accatggctt cgtacccctg ccatcaacac
      721 gcgtctgcgt tcgaccaggc tgcgcgttct cgcggccata gcaaccgacg tacggcgttg
      781 cgccctcgcc ggcagcaaga agccacggaa gtccgcctgg agcagaaaat gcccacgcta
      841 ctgcgggttt atatagacgg tcctcacggg atggggaaaa ccaccaccac gcaactgctg
      901 gtggccctgg gttcgcgcga cgatatcgtc tacgtacccg agccgatgac ttactggcag
      961 gtgctggggg cttccgagac aatcgcgaac atctacacca cacaacaccg cctcgaccag
     1021 ggtgagatat cggccgggga cgcggcggtg gtaatgacaa gcgcccagat aacaatgggc
     1081 atgccttatg ccgtgaccga cgccgttctg gctcctcata tcggggggga ggctgggagc
     1141 tcacatgccc cgcccccggc cctcaccctc atcttcgacc gccatcccat cgccgccctc
     1201 ctgtgctacc cggccgcgcg ataccttatg ggcagcatga ccccccaggc cgtgctggcg
     1261 ttcgtggccc tcatcccgcc gaccttgccc ggcacaaaca tcgtgttggg ggcccttccg
     1321 gaggacagac acatcgaccg cctggccaaa cgccagcgcc ccggcgagcg gcttgacctg
     1381 gctatgctgg ccgcgattcg ccgcgtttac gggctgcttg ccaatacggt gcggtatctg
     1441 cagggcggcg ggtcgtggcg ggaggattgg ggacagcttt cggggacggc cgtgccgccc
     1501 cagggtgccg agccccagag caacgcgggc ccacgacccc atatcgggga cacgttattt
     1561 accctgtttc gggcccccga gttgctggcc cccaacggcg acctgtacaa cgtgtttgcc
     1621 tgggccttgg acgtcttggc caaacgcctc cgtcccatgc acgtctttat cctggattac
     1681 gaccaatcgc ccgccggctg ccgggacgcc ctgctgcaac ttacctccgg gatggtccag
     1741 acccacgtca ccacccccgg ctccataccg acgatctgcg acctggcgcg cacgtttgcc
     1801 cgggagatgg gggaggctaa ctgaggatcc cacgtcacta ttgtatactc tatattatac
     1861 tctatgttat actctgtaat cctactcaat aaacgtgtca cgcctgtgaa accgtactaa
     1921 gtctcccgtg tattcttatc accatcaggt gacatcctcg cccaggctgt caatcatgcc
     1981 ggtatcgatt ccagtagcac cggccccacg ctgacaaccc actcttgcag cgttagcagc
     2041 gcccctctta acaagccgac ccccaccagc gtcgcggtta ctaacactcc tctccccgac
     2101 ctgcagccca gctccttcgg gattcctcta ctgcctcctc tttctaatcc ccagattctg
     2161 atatgcatcc ttgtcctaca gcgaggcagc atggcatgag gtcagaacac cagttctgga
     2221 gccagactgt ctaggttcac agcctgccat ttaccggcca tgtgactttg gcaagtttct
     2281 tagtctctct tgcctcactt tcctcatatg taaaatggga ataataatag tgcctacctc
     2341 agaaggttga tgtgaggaat gaaggtattg atacatgtaa acttagagca gtgtgggtac
     2401 aaaataaaca tgatgcaagt gttcaatcac tgtttttggg agaatgccat attctttaag
     2461 ccgttaaaga agaaaaaatg attaagaata atttcaaagt aatgcatgtt tcaagggcta
     2521 atgccaggtt gctcccagag tggtctctcc cagtgtctag aaattttaac atcttatgaa
     2581 aatgatatat atggtcaaaa atgtatttaa cctttccctt ggctgccttc cagggccaga
     2641 ggaagatctt ccaaggaaga tggaatactt ggaatttgtt tgtcatgcac cttctgagta
     2701 tttcaagtca cggtcatcac catttcccac agttcccacc agaccagaga agggctacat
     2761 atggactcat gttgggccta ctcctgcaat aacaattaag gaatcagttg ccaaccacct
     2821 agggagcgga ggaggttccg gtggaggtgg ttctggagat tacaaggatg acgacgataa
     2881 gggcgattac aaggatgacg acgataaggg agattacaag gatgacgacg ataaggtttc
     2941 aggaagcgga gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc
     3001 tggacctgga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3061 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3121 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3181 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3241 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3301 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3361 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3421 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     3481 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     3541 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     3601 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     3661 ggccgaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     3721 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     3781 cgccttctat cgccttcttg acgagttctt ctgattcgaa catcactggg tttccaggcc
     3841 tggtgtggtg gctcacgcct gtagccccag ctattgcacc actgctctcc aagctgggca
     3901 atggagtcag attctctttc ttaaaaaacc acaaaaaaac tggatttcca gttctctaat
     3961 attcttagta ccacaagata tgtcataggt atctttaaat gaaattctta gctggaaaag
     4021 tgactaaaaa gtttttctcc tgctacctag taataaacaa atcattgttt attactggtc
     4081 acttagaaaa ttaaaaggga tagggccagg cacagtggct tatgcctgta attgcagcac
     4141 ttttagaggc cgaggcaggc ggatcacctg aggtcgggaa gtggatcgcc tgaggtcagg
     4201 agttcgagac cagcctggcc aacatggcga aaccccgtcg ctactaaaaa tacaaaaatt
     4261 agccaggtgt ggtggcatgt gcctgtaatc ccagctattt gggaggctga ggcaggagaa
     4321 tcgcctaaac ccaggaggtg gaggttgtag tgagccaaga ttgcaccgct gtgctccagc
     4381 ctgggcaaca gagtgagact cttgtctcgg aaaaaaactt ggaaagtcct ctccactgac
     4441 tgtagcctcc aattcactgg ccgtcgttca ctggccgtcg ttttacaacg tcgtgactgg
     4501 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     4561 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     4621 gaatgcgatt tattcaacaa agccgccgtc ccgtcaagtc agcgtaatgc tctgccagtg
     4681 ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa
     4741 tatattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     4801 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     4861 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     4921 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc
     4981 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     5041 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa
     5101 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     5161 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     5221 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     5281 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     5341 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     5401 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     5461 atccatgttg gaatttaatc gcggcttcga gcaagacgtt tcccgttgaa tatggctcat
     5521 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt
     5581 tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa
     5641 tcgaactttt gctgagttga aggatcagat cacgcatctt cccgacaacg cagaccgttc
     5701 cgtggcaaag caaaagttca aaatcaccaa ctggtccacc tacaacaaag ctctcatcaa
     5761 ccgtggctcc ctcactttct ggctggatga tggggcgatt caggcctggt atgagtcagc
     5821 aacaccttct tcacgaggca gacctctcga cggagttcca ctgagcgtca gaccccgtag
     5881 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5941 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6001 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6061 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6121 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6181 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6241 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6301 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6361 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6421 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6481 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6541 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     6601 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     6661 aagcggaaga agctcgcaca ttcagcagcg tttttcagcg cgttttcgat caacgtttca
     6721 atgttggtat caacaccagg tttaactttg aacttatcgg cactgacggt tactgatttt
     6781 gaacttttgc tttgccacgg aacggtctgc gttgtcggga agatgcgtga tctgatcctt
     6841 caactcagca aaagt
//
