LOCUS       sequence-170753-        6970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human ZSCAN9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6970)
  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 6970)
  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_ZSCAN9
FEATURES             Location/Qualifiers
     source          1..6970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      24..612
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     513..532
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     766..783
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1083..1104
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1119..1124,1125..1142,1143..1149)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1157..1173
                     /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     1162..1184
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1181..1197
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1181..1197
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1374..1576
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1527..1547
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1573..1597
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1673..2803
                     /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             3837..3860
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3864..3887
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3891..3914
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3930..3986
                     /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             3999..4793
                     /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(4053..4072)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4663..4682
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5564..5580)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5573..5595)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(5799..6614)
                     /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     6522..6541
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6606..6625
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
       61 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
      121 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
      181 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
      241 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
      301 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
      361 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
      421 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
      481 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
      541 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
      601 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
      661 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
      721 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
      781 gaggaagcgg aagaagctcg cacattcagc agcgtttttc agcgcgtttt cgatcaacgt
      841 ttcaatgttg gtatcaacac caggtttaac tttgaactta tcggcactga cggttactga
      901 ttttgaactt ttgctttgcc acggaacggt ctgcgttgtc gggaagatgc gtgatctgat
      961 ccttcaactc agcaaaagtt cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     1021 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     1081 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     1141 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     1201 gattacgcca agcttgcatg cctgcaggtc gactctagag gatccccggg taccgagctc
     1261 gaattagctt gcatgcctgc aggtcaattc cctggcatta tgcccagtac atgaccttat
     1321 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     1381 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttcaagtct
     1441 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     1501 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     1561 ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg
     1621 ttttgacctc cataggagac accgggaccg atccagcctg gggatcccga ccatggcttc
     1681 gtacccctgc catcaacacg cgtctgcgtt cgaccaggct gcgcgttctc gcggccatag
     1741 caaccgacgt acggcgttgc gccctcgccg gcagcaagaa gccacggaag tccgcctgga
     1801 gcagaaaatg cccacgctac tgcgggttta tatagacggt cctcacggga tggggaaaac
     1861 caccaccacg caactgctgg tggccctggg ttcgcgcgac gatatcgtct acgtacccga
     1921 gccgatgact tactggcagg tgctgggggc ttccgagaca atcgcgaaca tctacaccac
     1981 acaacaccgc ctcgaccagg gtgagatatc ggccggggac gcggcggtgg taatgacaag
     2041 cgcccagata acaatgggca tgccttatgc cgtgaccgac gccgttctgg ctcctcatat
     2101 cgggggggag gctgggagct cacatgcccc gcccccggcc ctcaccctca tcttcgaccg
     2161 ccatcccatc gccgccctcc tgtgctaccc ggccgcgcga taccttatgg gcagcatgac
     2221 cccccaggcc gtgctggcgt tcgtggccct catcccgccg accttgcccg gcacaaacat
     2281 cgtgttgggg gcccttccgg aggacagaca catcgaccgc ctggccaaac gccagcgccc
     2341 cggcgagcgg cttgacctgg ctatgctggc cgcgattcgc cgcgtttacg ggctgcttgc
     2401 caatacggtg cggtatctgc agggcggcgg gtcgtggcgg gaggattggg gacagctttc
     2461 ggggacggcc gtgccgcccc agggtgccga gccccagagc aacgcgggcc cacgacccca
     2521 tatcggggac acgttattta ccctgtttcg ggcccccgag ttgctggccc ccaacggcga
     2581 cctgtacaac gtgtttgcct gggccttgga cgtcttggcc aaacgcctcc gtcccatgca
     2641 cgtctttatc ctggattacg accaatcgcc cgccggctgc cgggacgccc tgctgcaact
     2701 tacctccggg atggtccaga cccacgtcac cacccccggc tccataccga cgatctgcga
     2761 cctggcgcgc acgtttgccc gggagatggg ggaggctaac tgaggatccc acgtcactat
     2821 tgtatactct atattatact ctatgttata ctctgtaatc ctactcaata aacgtgtcac
     2881 gcctgtgaaa ccgtactaag tctcccgtgt attcttatca ccatcaggtg acatcctcgc
     2941 ccaggctgtc aatcatgccg gtatcgattc cagtagcacc ggccccacgc tgacaaccca
     3001 ctcttgcagc gttagcagcg cccctcttaa caagccgacc cccaccagcg tcgcggttac
     3061 taacactcct ctccccgacc tgcagcccag ctcttccttt tctatgatat atttttatag
     3121 acatagtcac cttcccatag gctcagggaa caagtgacat ttacctagcc tttttctttg
     3181 tttcagatga agtaaccaag actgaggaca gagagttggt gctaaggaaa gactgtccta
     3241 agatagtgga accacatggg aaaatgttta atgagcagac ctgggaggta tcacagcagg
     3301 atccctcaca tggagaagtt gttgaacata aggataggat agagaggcag tggggaaacc
     3361 tcttaggaga ggggcaacac aaatgtgatg aatgtgggaa gagctttact cagagctcag
     3421 gtctcattcg acatcaaaga attcatactg gagaaagacc ttatgaatgt aatgaatgtg
     3481 ggaaagcctt cagtcgaagt tctggtcttt ttaatcaccg aggaatccac aatatacaga
     3541 aacggtacca ctgcaaggag tgtgggaagg tcttcagtca gagtgcgggt cttatccagc
     3601 atcagagaat ccacaaagga gaaaagccgt atcagtgcag ccagtgcagt aagagctaca
     3661 gtcggcgttc atttctcatt gaacatcaga gaagccacac aggggagcga cctcaccagt
     3721 gcattgaatg tgggaaaagc tttaatcgac actgcaacct cattcgccat cagaagatcc
     3781 acacagtggc tgagctggta gggagcggag gaggttccgg tggaggtggt tctggagatt
     3841 acaaggatga cgacgataag ggcgattaca aggatgacga cgataaggga gattacaagg
     3901 atgacgacga taaggtttca ggaagcggag ctactaactt cagcctgctg aagcaggctg
     3961 gagacgtgga ggagaaccct ggacctggat cgtttcgcat gattgaacaa gatggattgc
     4021 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4081 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4141 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4201 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4261 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4321 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4381 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4441 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4501 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4561 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4621 actgtggccg gctgggtgtg gccgaccgct atcaggacat agcgttggct acccgtgata
     4681 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4741 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgattcgaac
     4801 atcgcttggc tatgagcaag ttttccagat caccacccaa gttgtgtggg gcaggttgag
     4861 actagaaaat gcctctttct tcctttctcc atgaaatgtg tttgaaacaa atcctgactt
     4921 aaggcccagg gacttcctta aaggaaagtt gggtgtttga agctactgtt ttctcttttg
     4981 ttcattttac ctctttctta ctcttactag ctgtgtccct cttatttata atttatttat
     5041 ttttttgaga tggctgctaa acccttctaa taatataata aatggcactg ccacagccaa
     5101 catgcctgaa agatttggta tctgtgttta actacaaagg ataacagagt tctactgatg
     5161 gtttataaca attgggtctt tggtgttccc tgtgaccagg gtctacacta taaacacctt
     5221 ttctacctta ctgcttctct ctgacaccta cgtgactctc ccttggaaag tgaccagcat
     5281 tggctgtgtg tttcagtgtc tgcagtgtgt gtccgcttac ttaaccatag ttggtcctca
     5341 caacagccac acagggaaat gagcgttatc attatcgcat tttacagatg aggactccaa
     5401 ggcttagaaa atccaagcat cagagccaag gctgaaaccc tgaactttta tttttagtaa
     5461 tgctttaatt ttggaataat atttgactta cagaaatgtt gcagagatgg acttggaaag
     5521 tcctctccac tgactgtagc ctccaattca ctggccgtcg ttcactggcc gtcgttttac
     5581 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     5641 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     5701 gcagcctgaa tggcgaatgc gatttattca acaaagccgc cgtcccgtca agtcagcgta
     5761 atgctctgcc agtgttacaa ccaattaacc aattctgatt agaaaaactc atcgagcatc
     5821 aaatgaaact gcaatatatt catatcagga ttatcaatac catatttttg aaaaagccgt
     5881 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     5941 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     6001 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     6061 agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     6121 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     6181 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     6241 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     6301 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     6361 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     6421 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     6481 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     6541 ccatataaat cagcatccat gttggaattt aatcgcggct tcgagcaaga cgtttcccgt
     6601 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt
     6661 catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg
     6721 ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc agatcacgca tcttcccgac
     6781 aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc cacctacaac
     6841 aaagctctca tcaaccgtgg ctccctcact ttctggctgg atgatggggc gattcaggcc
     6901 tggtatgagt cagcaacacc ttcttcacga ggcagacctc tcgacggagt tccactgagc
     6961 gtcagacccc
//
