LOCUS       sequence-245629-        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"
     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(2473..2489)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2482..2504)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2708..3523)
                     /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     3431..3450
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3515..3534
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3903..4491
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4392..4411
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4645..4662
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4962..4983
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4998..5003,5004..5021,5022..5028)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5036..5052
                     /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     5041..5063
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5060..5076
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5060..5076
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5253..5455
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5406..5426
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5452..5476
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5552..6682
                     /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 atgaatgggt taggaaatac tgaggagaaa atggacttgg aagagaatcc tgatgagcag
       61 tccgaaataa gagatatgtt tgttgaaatg ctggatgatt ttagggacaa tcattaccag
      121 ataaacagta tccaaaaaaa gcagttattt aaacattctg cctgcccttt tcgatgtcct
      181 aattgtggcc agcgttttga aactgaaaat ctagtggttg aacatatgtc tagctgctta
      241 gatcaagata tgtttaagag tgccatcatg gaagaaaatg aaagagatca cagacgaaag
      301 catttttgta atctgtgtgg aaaaggattt tatcagcggt gtcacttaag agaacactat
      361 actgttcata ctaaggaaaa acagtttgtt tgtcaaacat gtggaaagca gtttttaaga
      421 gagcgtcagt tgcgactgca caatgatatg cacaaaggca tggccaggta tgtctgttcc
      481 atttgtgatc aaggaaactt cagaaaacat gaccatgtac ggcatatgat ttctcattta
      541 tctgctggtg agactatatg ccaggtctgc tttcagatat tcccaaataa tgaacagttg
      601 gagcagcaca tggatgttca tctgtataca tgtggaatat gtggagcaaa atttaatttg
      661 aggaaagata tgagatcaca ttataatgcc aagcatttga aaagaaccgg 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 ctgattttct actgtactaa tgtttagatg
     1741 atagcagata aaacaccaaa gcaaaggata tgagctattt aggaattgat tatataagat
     1801 gatttgttag aaacaaattt caaggccctt ttaactttaa tatttttgtt taggatttta
     1861 agtatctaca tttaggtatt aaatgtttat catttttgtt gtttcttaat agaattattt
     1921 gtttttagtt tttcttagct atgattaaaa tttttaaatg tagactacaa gtggttgtta
     1981 cccattcaat gactattaaa ctttagtttt tcatcaataa ggtgatgact tcactatttc
     2041 tatgtgtttt tttttttttt aaggttatcc tgtgaaattt taaacccaga atcattggcc
     2101 atttctgttt aaattttaaa aattccttaa gtaattattt gaaactatcc cgtttgcttt
     2161 tagtgagtta actactcttt attccccctt attaatgaaa ttcatattct taaattgaca
     2221 agcttattag gcaagttagg tgcactgaat ctaaccttta aggttgacat gggctcaaac
     2281 ttggcctaaa aagattgatg aacctgagga aatttttata aatgaatatt tcctataatt
     2341 gataaaatat tatcatttaa ttatcacatt taaaacttat attaagtgta aattccagtg
     2401 gctttacctc acttgaaaaa cttggaaagt cctctccact gactgtagcc tccaattcac
     2461 tggccgtcgt tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     2521 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     2581 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgcg atttattcaa
     2641 caaagccgcc gtcccgtcaa gtcagcgtaa tgctctgcca gtgttacaac caattaacca
     2701 attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
     2761 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     2821 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     2881 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     2941 tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag acttgttcaa
     3001 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     3061 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     3121 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     3181 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
     3241 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     3301 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     3361 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     3421 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     3481 atcgcggctt cgagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac
     3541 tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt
     3601 aacatcagag attttgagac acaacgtggc tttgttgaat aaatcgaact tttgctgagt
     3661 tgaaggatca gatcacgcat cttcccgaca acgcagaccg ttccgtggca aagcaaaagt
     3721 tcaaaatcac caactggtcc acctacaaca aagctctcat caaccgtggc tccctcactt
     3781 tctggctgga tgatggggcg attcaggcct ggtatgagtc agcaacacct tcttcacgag
     3841 gcagacctct cgacggagtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3901 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3961 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4021 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4081 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4141 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4201 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4261 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4321 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4381 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4441 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4501 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4561 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     4621 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agaagctcgc
     4681 acattcagca gcgtttttca gcgcgttttc gatcaacgtt tcaatgttgg tatcaacacc
     4741 aggtttaact ttgaacttat cggcactgac ggttactgat tttgaacttt tgctttgcca
     4801 cggaacggtc tgcgttgtcg ggaagatgcg tgatctgatc cttcaactca gcaaaagttc
     4861 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     4921 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     4981 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     5041 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcttgcatgc
     5101 ctgcaggtcg actctagagg atccccgggt accgagctcg aattagcttg catgcctgca
     5161 ggtcaattcc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     5221 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     5281 ggcgtggata gcggtttgac tcacggggat ttcaagtctc caccccattg acgtcaatgg
     5341 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     5401 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt
     5461 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc ataggagaca
     5521 ccgggaccga tccagcctgg ggatcccgac catggcttcg tacccctgcc atcaacacgc
     5581 gtctgcgttc gaccaggctg cgcgttctcg cggccatagc aaccgacgta cggcgttgcg
     5641 ccctcgccgg cagcaagaag ccacggaagt ccgcctggag cagaaaatgc ccacgctact
     5701 gcgggtttat atagacggtc ctcacgggat ggggaaaacc accaccacgc aactgctggt
     5761 ggccctgggt tcgcgcgacg atatcgtcta cgtacccgag ccgatgactt actggcaggt
     5821 gctgggggct tccgagacaa tcgcgaacat ctacaccaca caacaccgcc tcgaccaggg
     5881 tgagatatcg gccggggacg cggcggtggt aatgacaagc gcccagataa caatgggcat
     5941 gccttatgcc gtgaccgacg ccgttctggc tcctcatatc gggggggagg ctgggagctc
     6001 acatgccccg cccccggccc tcaccctcat cttcgaccgc catcccatcg ccgccctcct
     6061 gtgctacccg gccgcgcgat accttatggg cagcatgacc ccccaggccg tgctggcgtt
     6121 cgtggccctc atcccgccga ccttgcccgg cacaaacatc gtgttggggg cccttccgga
     6181 ggacagacac atcgaccgcc tggccaaacg ccagcgcccc ggcgagcggc ttgacctggc
     6241 tatgctggcc gcgattcgcc gcgtttacgg gctgcttgcc aatacggtgc ggtatctgca
     6301 gggcggcggg tcgtggcggg aggattgggg acagctttcg gggacggccg tgccgcccca
     6361 gggtgccgag ccccagagca acgcgggccc acgaccccat atcggggaca cgttatttac
     6421 cctgtttcgg gcccccgagt tgctggcccc caacggcgac ctgtacaacg tgtttgcctg
     6481 ggccttggac gtcttggcca aacgcctccg tcccatgcac gtctttatcc tggattacga
     6541 ccaatcgccc gccggctgcc gggacgccct gctgcaactt acctccggga tggtccagac
     6601 ccacgtcacc acccccggct ccataccgac gatctgcgac ctggcgcgca cgtttgcccg
     6661 ggagatgggg gaggctaact gaggatccca cgtcactatt gtatactcta tattatactc
     6721 tatgttatac tctgtaatcc tactcaataa acgtgtcacg cctgtgaaac cgtactaagt
     6781 ctcccgtgta ttcttatcac catcaggtga catcctcgcc caggctgtca atcatgccgg
     6841 tatcgattcc agtagcaccg gccccacgct gacaacccac tcttgcagcg ttagcagcgc
     6901 ccctcttaac aagccgaccc ccaccagcgt cgcggttact aacactcctc tccccgacct
     6961 gcagcccagc t
//
