LOCUS       sequence-271137-        8308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human ZBTB7B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8308)
  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 8308)
  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_ZBTB7B
FEATURES             Location/Qualifiers
     source          1..8308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..203
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     154..174
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     200..224
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             300..1430
                     /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             2464..2487
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2491..2514
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2518..2541
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2557..2613
                     /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             2626..3420
                     /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(2680..2699)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3290..3309
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4191..4207)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4200..4222)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(5764..6579)
                     /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     6487..6506
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6571..6590
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6959..7547
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7448..7467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7701..7718
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8018..8039
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8054..8059,8060..8077,8078..8084)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8092..8108
                     /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     8097..8119
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8116..8132
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8116..8132
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      181 gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
      241 cacgctgttt tgacctccat aggagacacc gggaccgatc cagcctgggg atcccgacca
      301 tggcttcgta cccctgccat caacacgcgt ctgcgttcga ccaggctgcg cgttctcgcg
      361 gccatagcaa ccgacgtacg gcgttgcgcc ctcgccggca gcaagaagcc acggaagtcc
      421 gcctggagca gaaaatgccc acgctactgc gggtttatat agacggtcct cacgggatgg
      481 ggaaaaccac caccacgcaa ctgctggtgg ccctgggttc gcgcgacgat atcgtctacg
      541 tacccgagcc gatgacttac tggcaggtgc tgggggcttc cgagacaatc gcgaacatct
      601 acaccacaca acaccgcctc gaccagggtg agatatcggc cggggacgcg gcggtggtaa
      661 tgacaagcgc ccagataaca atgggcatgc cttatgccgt gaccgacgcc gttctggctc
      721 ctcatatcgg gggggaggct gggagctcac atgccccgcc cccggccctc accctcatct
      781 tcgaccgcca tcccatcgcc gccctcctgt gctacccggc cgcgcgatac cttatgggca
      841 gcatgacccc ccaggccgtg ctggcgttcg tggccctcat cccgccgacc ttgcccggca
      901 caaacatcgt gttgggggcc cttccggagg acagacacat cgaccgcctg gccaaacgcc
      961 agcgccccgg cgagcggctt gacctggcta tgctggccgc gattcgccgc gtttacgggc
     1021 tgcttgccaa tacggtgcgg tatctgcagg gcggcgggtc gtggcgggag gattggggac
     1081 agctttcggg gacggccgtg ccgccccagg gtgccgagcc ccagagcaac gcgggcccac
     1141 gaccccatat cggggacacg ttatttaccc tgtttcgggc ccccgagttg ctggccccca
     1201 acggcgacct gtacaacgtg tttgcctggg ccttggacgt cttggccaaa cgcctccgtc
     1261 ccatgcacgt ctttatcctg gattacgacc aatcgcccgc cggctgccgg gacgccctgc
     1321 tgcaacttac ctccgggatg gtccagaccc acgtcaccac ccccggctcc ataccgacga
     1381 tctgcgacct ggcgcgcacg tttgcccggg agatggggga ggctaactga ggatcccacg
     1441 tcactattgt atactctata ttatactcta tgttatactc tgtaatccta ctcaataaac
     1501 gtgtcacgcc tgtgaaaccg tactaagtct cccgtgtatt cttatcacca tcaggtgaca
     1561 tcctcgccca ggctgtcaat catgccggta tcgattccag tagcaccggc cccacgctga
     1621 caacccactc ttgcagcgtt agcagcgccc ctcttaacaa gccgaccccc accagcgtcg
     1681 cggttactaa cactcctctc cccgacctgc agcccagctc cagcaggggg tggatctgga
     1741 gcatggagga ttcggtgccc aggagggcag tgctggaagc agaggagtgg ataatgacaa
     1801 ggcctagtta agctagaggt gagctagcag ggtatctcct ggggcaatag tggggtaaca
     1861 aatggtgagg aacagggatg ggacaaggcc agggtgggat gaccaggagg agccagggat
     1921 cccatcctga acacctccct gcccctcacc cctgcctgtg ccaggaacga caagctgaag
     1981 atccacatgc ggaagcacac gggagagcgc ccctactcat gcccgcactg cccagcccgc
     2041 ttcctgcaca gctacgacct caagaaccac atgcacctgc acacagggga ccggccctat
     2101 gagtgccacc tgtgccacaa ggctttcgcc aaggaggacc acctgcagcg ccacctcaaa
     2161 ggccagaact gcctggaggt gcgcacccga cggcgccgca aggacgatgc accaccccac
     2221 tacccaccac cctctaccgc tgctgcatcc cccgctggcc tcgacctctc caatggccac
     2281 ctggacacct tccgcctctc tctagctcga ttctgggagc agtcagcccc cactgggccc
     2341 ccggtctcta ccccagggcc ccctgatgac gatgaggagg aaggggcacc caccacaccc
     2401 caggctgaag gtgccatgga gtcctctggg agcggaggag gttccggtgg aggtggttct
     2461 ggagattaca aggatgacga cgataagggc gattacaagg atgacgacga taagggagat
     2521 tacaaggatg acgacgataa ggtttcagga agcggagcta ctaacttcag cctgctgaag
     2581 caggctggag acgtggagga gaaccctgga cctggatcgt ttcgcatgat tgaacaagat
     2641 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2701 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2761 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     2821 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     2881 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     2941 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3001 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3061 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3121 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     3181 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3241 ttcatcgact gtggccggct gggtgtggcc gaccgctatc aggacatagc gttggctacc
     3301 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3361 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3421 ttcgaacatc cgagggccag actgaagcag cacaaggccg gggacaccca tgccaagcag
     3481 tgggagcacg caggacagac acagcagggg tctggggcac ggagccttgc tggcatcagc
     3541 atcagccctt cctcccagag ccctcattcc aattccaagc taagaaggta ttggggcaga
     3601 ggctccccaa attggggtga tcccccaagg agtgatacat atattgtgta tatatttaca
     3661 gctgtattgt aaaagtgggg tccctgtccc cagctgctcc tggggagtag aagcaataat
     3721 gtatttctaa tttgtgggtc ccacttcggc tatgcgggtt tctagggggt gggggcttgg
     3781 gaccaaagcc ttgccccgcc cctatgcccc ttgggggttt tggctgtgta agggggtgaa
     3841 ggactgcccc tccctttcga gacccctcct tcctggtttc tgttcctttt tcctggcagt
     3901 gaattatgca aagggggccg gcaaaggaag ggtaggtggg ggaaagccag gtggaagctt
     3961 gaaagactgg gggactgggc ctgtaaggaa ggagccatcc cagtccccct ccgccctgct
     4021 cccggcgctg agtcatgggg tcgtggagaa gggggcgggg tggcctgatt ggctcgcctg
     4081 cccctggggg cagtagaggg gccccgccca gctaggggag ccgctccgtt ccactcccct
     4141 tggaaagtcc tctccactga ctgtagcctc caattcactg gccgtcgttc actggccgtc
     4201 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     4261 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     4321 cagttgcgca gccctgatga atcccctaat gattttggta aaaatcatta agttaaggtg
     4381 gatacacatc ttgtcatatg atcaaatggt ttcgcgaaaa atcaataatc agacaacaag
     4441 atgtgcgaac tcgatatttt acacgactct ctttaccaat tctgccccga attacactta
     4501 aaacgactca acagcttaac gttggcttgc cacgcattac ttgactgtaa aactctcact
     4561 cttaccgaac ttggccgtaa cctgccaacc aaagcgagaa caaaacataa catcaaacga
     4621 atcgaccgat tgttaggtaa tcgtcacctc cacaaagagc gactcgctgt ataccgttgg
     4681 catgctagct ttatctgttc gggcaatacg atgcccattg tacttgttga ctggtctgat
     4741 attcgtgagc aaaaacgact tatggtattg cgagcttcag tcgcactaca cggtcgttct
     4801 gttactcttt atgagaaagc gttcccgctt tcagagcaat gttcaaagaa agctcatgac
     4861 caatttctag ccgaccttgc gagcattcta ccgagtaaca ccacaccgct cattgtcagt
     4921 gatgctggct ttaaagtgcc atggtataaa tccgttgaga agctgggttg gtactggtta
     4981 agtcgagtaa gaggaaaagt acaatatgca gacctaggag cggaaaactg gaaacctatc
     5041 agcaacttac atgatatgtc atctagtcac tcaaagactt taggctataa gaggctgact
     5101 aaaagcaatc caatctcatg ccaaattcta ttgtataaat ctcgctctaa aggccgaaaa
     5161 aatcagcgct cgacacggac tcattgtcac cacccgtcac ctaaaatcta ctcagcgtcg
     5221 gcaaaggagc catgggttct agcaactaac ttacctgttg aaattcgaac acccaaacaa
     5281 cttgttaata tctattcgaa gcgaatgcag attgaagaaa ccttccgaga cttgaaaagt
     5341 cctgcctacg gactaggcct acgccatagc cgaacgagca gctcagagcg ttttgatatc
     5401 atgctgctaa tcgccctgat gcttcaacta acatgttggc ttgcgggcgt tcatgctcag
     5461 aaacaaggtt gggacaagca cttccaggct aacacagtca gaaatcgaaa cgtactctca
     5521 acagttcgct taggcatgga agttttgcgg cattctggct acacaataac aagggaagac
     5581 ttactcgtgg ctgcaaccct actagctcaa aatttattca cacatggtta cgctttgggg
     5641 aaattatgag gggatctctc agtgcgcagc ctgaatggcg aatgcgattt attcaacaaa
     5701 gccgccgtcc cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc
     5761 tgattagaaa aactcatcga gcatcaaatg aaactgcaat atattcatat caggattatc
     5821 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     5881 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     5941 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     6001 gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg
     6061 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     6121 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     6181 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     6241 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     6301 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     6361 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     6421 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     6481 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     6541 cggcttcgag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     6601 tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca
     6661 tcagagattt tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa
     6721 ggatcagatc acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa
     6781 aatcaccaac tggtccacct acaacaaagc tctcatcaac cgtggctccc tcactttctg
     6841 gctggatgat ggggcgattc aggcctggta tgagtcagca acaccttctt cacgaggcag
     6901 acctctcgac ggagttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6961 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     7021 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     7081 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     7141 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     7201 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     7261 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     7321 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     7381 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     7441 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     7501 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     7561 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     7621 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     7681 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagaa gctcgcacat
     7741 tcagcagcgt ttttcagcgc gttttcgatc aacgtttcaa tgttggtatc aacaccaggt
     7801 ttaactttga acttatcggc actgacggtt actgattttg aacttttgct ttgccacgga
     7861 acggtctgcg ttgtcgggaa gatgcgtgat ctgatccttc aactcagcaa aagttcccaa
     7921 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     7981 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     8041 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     8101 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt gcatgcctgc
     8161 aggtcgactc tagaggatcc ccgggtaccg agctcgaatt agcttgcatg cctgcaggtc
     8221 aattccctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     8281 ctacgtatta gtcatcgcta ttaccatg
//
