LOCUS       sequence-171196-        6795 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human ZFP1_iso1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6795)
  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 6795)
  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_ZFP1_iso1
FEATURES             Location/Qualifiers
     source          1..6795
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      16..604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     505..524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     758..775
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1075..1096
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1111..1116,1117..1134,1135..1141)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1149..1165
                     /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     1154..1176
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1173..1189
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1173..1189
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1366..1568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1519..1539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1565..1589
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1665..2795
                     /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             3654..3677
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3681..3704
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3708..3731
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3747..3803
                     /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             3816..4610
                     /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(3870..3889)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4480..4499
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5381..5397)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5390..5412)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(5616..6431)
                     /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     6339..6358
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6423..6442
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
       61 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
      121 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
      181 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
      241 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
      301 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
      361 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
      421 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
      481 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
      541 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
      601 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
      661 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
      721 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      781 ggaagaagct cgcacattca gcagcgtttt tcagcgcgtt ttcgatcaac gtttcaatgt
      841 tggtatcaac accaggttta actttgaact tatcggcact gacggttact gattttgaac
      901 ttttgctttg ccacggaacg gtctgcgttg tcgggaagat gcgtgatctg atccttcaac
      961 tcagcaaaag ttcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     1021 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     1081 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     1141 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     1201 caagcttgca tgcctgcagg tcgactctag aggatccccg ggtaccgagc tcgaattagc
     1261 ttgcatgcct gcaggtcaat tccctggcat tatgcccagt acatgacctt atgggacttt
     1321 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     1381 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttcaagt ctccacccca
     1441 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     1501 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     1561 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     1621 tccataggag acaccgggac cgatccagcc tggggatccc gaccatggct tcgtacccct
     1681 gccatcaaca cgcgtctgcg ttcgaccagg ctgcgcgttc tcgcggccat agcaaccgac
     1741 gtacggcgtt gcgccctcgc cggcagcaag aagccacgga agtccgcctg gagcagaaaa
     1801 tgcccacgct actgcgggtt tatatagacg gtcctcacgg gatggggaaa accaccacca
     1861 cgcaactgct ggtggccctg ggttcgcgcg acgatatcgt ctacgtaccc gagccgatga
     1921 cttactggca ggtgctgggg gcttccgaga caatcgcgaa catctacacc acacaacacc
     1981 gcctcgacca gggtgagata tcggccgggg acgcggcggt ggtaatgaca agcgcccaga
     2041 taacaatggg catgccttat gccgtgaccg acgccgttct ggctcctcat atcggggggg
     2101 aggctgggag ctcacatgcc ccgcccccgg ccctcaccct catcttcgac cgccatccca
     2161 tcgccgccct cctgtgctac ccggccgcgc gataccttat gggcagcatg accccccagg
     2221 ccgtgctggc gttcgtggcc ctcatcccgc cgaccttgcc cggcacaaac atcgtgttgg
     2281 gggcccttcc ggaggacaga cacatcgacc gcctggccaa acgccagcgc cccggcgagc
     2341 ggcttgacct ggctatgctg gccgcgattc gccgcgttta cgggctgctt gccaatacgg
     2401 tgcggtatct gcagggcggc gggtcgtggc gggaggattg gggacagctt tcggggacgg
     2461 ccgtgccgcc ccagggtgcc gagccccaga gcaacgcggg cccacgaccc catatcgggg
     2521 acacgttatt taccctgttt cgggcccccg agttgctggc ccccaacggc gacctgtaca
     2581 acgtgtttgc ctgggccttg gacgtcttgg ccaaacgcct ccgtcccatg cacgtcttta
     2641 tcctggatta cgaccaatcg cccgccggct gccgggacgc cctgctgcaa cttacctccg
     2701 ggatggtcca gacccacgtc accacccccg gctccatacc gacgatctgc gacctggcgc
     2761 gcacgtttgc ccgggagatg ggggaggcta actgaggatc ccacgtcact attgtatact
     2821 ctatattata ctctatgtta tactctgtaa tcctactcaa taaacgtgtc acgcctgtga
     2881 aaccgtacta agtctcccgt gtattcttat caccatcagg tgacatcctc gcccaggctg
     2941 tcaatcatgc cggtatcgat tccagtagca ccggccccac gctgacaacc cactcttgca
     3001 gcgttagcag cgcccctctt aacaagccga cccccaccag cgtcgcggtt actaacactc
     3061 ctctccccga cctgcagccc agctatcaga gaattcacac tggggagaaa cctttcgagt
     3121 gtccggaatg tggaaaagct ttcacccacc agtcaaacct cattgtacac cagagagcac
     3181 atatggagaa gaagccctat gagtgcagtg aatgtggaaa gacatttgcc caaaagtttg
     3241 aactcaccac acaccagaga attcatacag gagagcgacc ctatgagtgt aacgaatgtg
     3301 caaaaacctt ctttaagaag tcaaacctta tcatacatca gaagattcac acgggggaga
     3361 aacgctatga gtgcagtgaa tgtggaaaat cctttatcca gaactcacag ctcatcatac
     3421 acatgagaac tcatacagga gagaaaccct atgaatgtac tgagtgcggc aaaactttca
     3481 gccagaggtc aactcttaga ttacacttgc gaatccacac aggagagaaa ccatatgagt
     3541 gttccgaatg tgggaaggcc tttagcagga agtcccgact cagtgtccat cagagagttc
     3601 acatcgggga gaaacccggg agcggaggag gttccggtgg aggtggttct ggagattaca
     3661 aggatgacga cgataagggc gattacaagg atgacgacga taagggagat tacaaggatg
     3721 acgacgataa ggtttcagga agcggagcta ctaacttcag cctgctgaag caggctggag
     3781 acgtggagga gaaccctgga cctggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3841 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3901 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3961 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     4021 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4081 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4141 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4201 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4261 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4321 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     4381 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4441 gtggccggct gggtgtggcc gaccgctatc aggacatagc gttggctacc cgtgatattg
     4501 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4561 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga ttcgaacatc
     4621 ctgtggaaaa ctcctgccag aactcttcaa gcgggtgaaa aacctcatga cagtattgag
     4681 ggaacatggg aattcatact gagatgcaat ctctcaactc aaaaatgtat taaaaatagg
     4741 atcccatgag aacattatac tggaagttac atgtgatacc cagctaaaga atacatatca
     4801 gaatatatcc agttgtaaat agccataccc agttgtacta caatgagctt tttaaaatct
     4861 tgaatgaatt gagtattcta gacagcagca taagaaatat acaaacagta tcctatgaat
     4921 ttagtggttt tatgtggata tgcaacaaaa cacaagtggt atgctttaga tctgcacatg
     4981 tgaatttagc ataatcactg ggaatttgaa attcttgtcc tctgtgataa ttaggacata
     5041 acaagatttt ccatactaaa catcacatgt gtttttcagt atctctgcaa tccagtatgc
     5101 attccaaatg aaatgtgtaa catttaaagt agcatggaac ctacgtcttt ccctactgct
     5161 tgctggcata tatcagttgg tatgaacatt gttcttgagc tgcctcttag gaaacagaag
     5221 acagtgttga agttttccct gcttctggtt tgctgaagat tttctagaag cactatttac
     5281 acaaatatgc aaatgtgaaa taaccgatct ataacgtgaa ggaggcagct tggaaagtcc
     5341 tctccactga ctgtagcctc caattcactg gccgtcgttc actggccgtc gttttacaac
     5401 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     5461 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     5521 gcctgaatgg cgaatgcgat ttattcaaca aagccgccgt cccgtcaagt cagcgtaatg
     5581 ctctgccagt gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa
     5641 tgaaactgca atatattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     5701 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     5761 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     5821 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc
     5881 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     5941 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     6001 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     6061 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     6121 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     6181 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     6241 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     6301 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     6361 tataaatcag catccatgtt ggaatttaat cgcggcttcg agcaagacgt ttcccgttga
     6421 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     6481 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt
     6541 tgttgaataa atcgaacttt tgctgagttg aaggatcaga tcacgcatct tcccgacaac
     6601 gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca actggtccac ctacaacaaa
     6661 gctctcatca accgtggctc cctcactttc tggctggatg atggggcgat tcaggcctgg
     6721 tatgagtcag caacaccttc ttcacgaggc agacctctcg acggagttcc actgagcgtc
     6781 agaccccgta gaaaa
//
