LOCUS       sequence-260780-        6971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human ZBTB1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6971)
  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 6971)
  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_ZBTB1
FEATURES             Location/Qualifiers
     source          1..6971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     100..117
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    417..438
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(453..458,459..476,477..483)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    491..507
                     /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     496..518
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     515..531
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     515..531
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        708..910
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     861..881
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     907..931
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1007..2137
                     /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             3171..3194
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3198..3221
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3225..3248
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3264..3320
                     /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             3333..4127
                     /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(3387..3406)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3997..4016
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4899..4915)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4908..4930)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(5134..5949)
                     /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     5857..5876
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5941..5960
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6329..6917
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6818..6837
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
       61 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
      121 gcggaagaag ctcgcacatt cagcagcgtt tttcagcgcg ttttcgatca acgtttcaat
      181 gttggtatca acaccaggtt taactttgaa cttatcggca ctgacggtta ctgattttga
      241 acttttgctt tgccacggaa cggtctgcgt tgtcgggaag atgcgtgatc tgatccttca
      301 actcagcaaa agttcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
      361 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
      421 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
      481 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
      541 gccaagcttg catgcctgca ggtcgactct agaggatccc cgggtaccga gctcgaatta
      601 gcttgcatgc ctgcaggtca attccctggc attatgccca gtacatgacc ttatgggact
      661 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      721 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcaa gtctccaccc
      781 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      841 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      901 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
      961 cctccatagg agacaccggg accgatccag cctggggatc ccgaccatgg cttcgtaccc
     1021 ctgccatcaa cacgcgtctg cgttcgacca ggctgcgcgt tctcgcggcc atagcaaccg
     1081 acgtacggcg ttgcgccctc gccggcagca agaagccacg gaagtccgcc tggagcagaa
     1141 aatgcccacg ctactgcggg tttatataga cggtcctcac gggatgggga aaaccaccac
     1201 cacgcaactg ctggtggccc tgggttcgcg cgacgatatc gtctacgtac ccgagccgat
     1261 gacttactgg caggtgctgg gggcttccga gacaatcgcg aacatctaca ccacacaaca
     1321 ccgcctcgac cagggtgaga tatcggccgg ggacgcggcg gtggtaatga caagcgccca
     1381 gataacaatg ggcatgcctt atgccgtgac cgacgccgtt ctggctcctc atatcggggg
     1441 ggaggctggg agctcacatg ccccgccccc ggccctcacc ctcatcttcg accgccatcc
     1501 catcgccgcc ctcctgtgct acccggccgc gcgatacctt atgggcagca tgacccccca
     1561 ggccgtgctg gcgttcgtgg ccctcatccc gccgaccttg cccggcacaa acatcgtgtt
     1621 gggggccctt ccggaggaca gacacatcga ccgcctggcc aaacgccagc gccccggcga
     1681 gcggcttgac ctggctatgc tggccgcgat tcgccgcgtt tacgggctgc ttgccaatac
     1741 ggtgcggtat ctgcagggcg gcgggtcgtg gcgggaggat tggggacagc tttcggggac
     1801 ggccgtgccg ccccagggtg ccgagcccca gagcaacgcg ggcccacgac cccatatcgg
     1861 ggacacgtta tttaccctgt ttcgggcccc cgagttgctg gcccccaacg gcgacctgta
     1921 caacgtgttt gcctgggcct tggacgtctt ggccaaacgc ctccgtccca tgcacgtctt
     1981 tatcctggat tacgaccaat cgcccgccgg ctgccgggac gccctgctgc aacttacctc
     2041 cgggatggtc cagacccacg tcaccacccc cggctccata ccgacgatct gcgacctggc
     2101 gcgcacgttt gcccgggaga tgggggaggc taactgagga tcccacgtca ctattgtata
     2161 ctctatatta tactctatgt tatactctgt aatcctactc aataaacgtg tcacgcctgt
     2221 gaaaccgtac taagtctccc gtgtattctt atcaccatca ggtgacatcc tcgcccaggc
     2281 tgtcaatcat gccggtatcg attccagtag caccggcccc acgctgacaa cccactcttg
     2341 cagcgttagc agcgcccctc ttaacaagcc gacccccacc agcgtcgcgg ttactaacac
     2401 tcctctcccc gacctgcagc ccagctatga atgggttagg aaatactgag gagaaaatgg
     2461 acttggaaga gaatcctgat gagcagtccg aaataagaga tatgtttgtt gaaatgctgg
     2521 atgattttag ggacaatcat taccagataa acagtatcca aaaaaagcag ttatttaaac
     2581 attctgcctg cccttttcga tgtcctaatt gtggccagcg ttttgaaact gaaaatctag
     2641 tggttgaaca tatgtctagc tgcttagatc aagatatgtt taagagtgcc atcatggaag
     2701 aaaatgaaag agatcacaga cgaaagcatt tttgtaatct gtgtggaaaa ggattttatc
     2761 agcggtgtca cttaagagaa cactatactg ttcatactaa ggaaaaacag tttgtttgtc
     2821 aaacatgtgg aaagcagttt ttaagagagc gtcagttgcg actgcacaat gatatgcaca
     2881 aaggcatggc caggtatgtc tgttccattt gtgatcaagg aaacttcaga aaacatgacc
     2941 atgtacggca tatgatttct catttatctg ctggtgagac tatatgccag gtctgctttc
     3001 agatattccc aaataatgaa cagttggagc agcacatgga tgttcatctg tatacatgtg
     3061 gaatatgtgg agcaaaattt aatttgagga aagatatgag atcacattat aatgccaagc
     3121 atttgaaaag aaccgggagc ggaggaggtt ccggtggagg tggttctgga gattacaagg
     3181 atgacgacga taagggcgat tacaaggatg acgacgataa gggagattac aaggatgacg
     3241 acgataaggt ttcaggaagc ggagctacta acttcagcct gctgaagcag gctggagacg
     3301 tggaggagaa ccctggacct ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3361 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3421 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3481 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     3541 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     3601 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     3661 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     3721 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3781 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3841 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     3901 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     3961 gccggctggg tgtggccgac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4021 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4081 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgattc gaacatctga
     4141 ttttctactg tactaatgtt tagatgatag cagataaaac accaaagcaa aggatatgag
     4201 ctatttagga attgattata taagatgatt tgttagaaac aaatttcaag gcccttttaa
     4261 ctttaatatt tttgtttagg attttaagta tctacattta ggtattaaat gtttatcatt
     4321 tttgttgttt cttaatagaa ttatttgttt ttagtttttc ttagctatga ttaaaatttt
     4381 taaatgtaga ctacaagtgg ttgttaccca ttcaatgact attaaacttt agtttttcat
     4441 caataaggtg atgacttcac tatttctatg tgtttttttt ttttttaagg ttatcctgtg
     4501 aaattttaaa cccagaatca ttggccattt ctgtttaaat tttaaaaatt ccttaagtaa
     4561 ttatttgaaa ctatcccgtt tgcttttagt gagttaacta ctctttattc ccccttatta
     4621 atgaaattca tattcttaaa ttgacaagct tattaggcaa gttaggtgca ctgaatctaa
     4681 cctttaaggt tgacatgggc tcaaacttgg cctaaaaaga ttgatgaacc tgaggaaatt
     4741 tttataaatg aatatttcct ataattgata aaatattatc atttaattat cacatttaaa
     4801 acttatatta agtgtaaatt ccagtggctt tacctcactt gaaaaacttg gaaagtcctc
     4861 tccactgact gtagcctcca attcactggc cgtcgttcac tggccgtcgt tttacaacgt
     4921 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     4981 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     5041 ctgaatggcg aatgcgattt attcaacaaa gccgccgtcc cgtcaagtca gcgtaatgct
     5101 ctgccagtgt tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg
     5161 aaactgcaat atattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     5221 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     5281 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     5341 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagctt
     5401 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     5461 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     5521 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     5581 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     5641 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     5701 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     5761 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     5821 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     5881 taaatcagca tccatgttgg aatttaatcg cggcttcgag caagacgttt cccgttgaat
     5941 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     6001 tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa cgtggctttg
     6061 ttgaataaat cgaacttttg ctgagttgaa ggatcagatc acgcatcttc ccgacaacgc
     6121 agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac tggtccacct acaacaaagc
     6181 tctcatcaac cgtggctccc tcactttctg gctggatgat ggggcgattc aggcctggta
     6241 tgagtcagca acaccttctt cacgaggcag acctctcgac ggagttccac tgagcgtcag
     6301 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6361 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6421 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6481 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6541 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6601 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6661 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6721 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6781 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6841 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6901 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6961 ggccttttgc t
//
