LOCUS       sequence-267404-        6970 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Donor vector for 3' FLAG tag of human ZHX3_iso1.
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 Apr 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_ZHX3_iso1
FEATURES             Location/Qualifiers
     source          1..6970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1045..1068
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1072..1095
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1099..1122
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1138..1194
                     /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             1207..2001
                     /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(1261..1280)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1871..1890
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2772..2788)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2781..2803)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(3007..3822)
                     /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     3730..3749
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3814..3833
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4202..4790
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4691..4710
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4944..4961
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5261..5282
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5297..5302,5303..5320,5321..5327)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5335..5351
                     /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     5340..5362
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5359..5375
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5359..5375
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5552..5754
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5705..5725
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5751..5775
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(5851..6970,1..11)
                     /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 aggctaactg aggatcccac gtcactattg tatactctat attatactct atgttatact
       61 ctgtaatcct actcaataaa cgtgtcacgc ctgtgaaacc gtactaagtc tcccgtgtat
      121 tcttatcacc atcaggtgac atcctcgccc aggctgtcaa tcatgccggt atcgattcca
      181 gtagcaccgg ccccacgctg acaacccact cttgcagcgt tagcagcgcc cctcttaaca
      241 agccgacccc caccagcgtc gcggttacta acactcctct ccccgacctg cagcccagct
      301 acatgttgct gtcctttgtg ggacagagca ggaatggcag ggggtggcca ttgccattgg
      361 cattgttctc ttctgtctga gacaggtaac tgtgggtaca ggaagggcca atctgtttat
      421 ctgtacctgg tttctcatgg ctttctcagc cagctgcaat taaggaccat ttctttatga
      481 atcacagatg atgaacctct ctgatatcag ccttactttg actttgatgt tagggttgca
      541 agttacaaat gaagtgaaga catcagtaga atacatatca gaactagacc cagctgctat
      601 tatattggaa cagtatgttc tggagagcct ggaaggagag ggcaggatga agcattttag
      661 gacaggactc cagagacggt ggtggcggaa gcctggggag ctagggggat ttagtgggcc
      721 tggagaggcc aaaccactgc tcctgctgct aggcctaggg tttagcaggg tctctcagag
      781 agtctgatgg ccttacaagg ttgagccagc atcctgtcct taacccacat tccctcaggc
      841 tggagggtgg gggagtgggt ggcagaaggc cattcattgc tcatggaatt aagaggtcct
      901 caaagccagt gatgccaccc tggtatagca aggaccaggc tgcctggggg gcaggtggtg
      961 gcagctgccg tagagtgaca ggcatgtgtt ttcttctcag aaacagacgg gagcggagga
     1021 ggttccggtg gaggtggttc tggagattac aaggatgacg acgataaggg cgattacaag
     1081 gatgacgacg ataagggaga ttacaaggat gacgacgata aggtttcagg aagcggagct
     1141 actaacttca gcctgctgaa gcaggctgga gacgtggagg agaaccctgg acctggatcg
     1201 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     1261 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     1321 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     1381 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     1441 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     1501 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     1561 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     1621 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     1681 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     1741 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     1801 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc cgaccgctat
     1861 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     1921 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     1981 cttcttgacg agttcttctg attcgaacat ctgaaggact ggccagtctg ggaaaccgcc
     2041 tgccacgtgg aagagccaaa cccgactctc tgctgccaca tgccgttccc atgcccggct
     2101 gctgggcacc tgggagagct tccagaatcc tcgcagacag cccagagcct gccgctaccc
     2161 tcggcctgcc caccaccaag caagcagcaa gcaagatggg gttctcatca gttcttcctc
     2221 ccacaatgta ggacctttcc tttaccttcc aatggataaa atagttcaga gttcatagtc
     2281 atattcatag acacagaatc aagcttttaa catatacatc cacctctata tgttaaataa
     2341 aacatcagat tatcaacact gtcattacgt agaaactttg gttagccaag cagtgcattg
     2401 tcagttacgt catctctaaa aatgacctgt gtctgttctc tggggattgc tgggtcacag
     2461 gtgcccctca ccttccacag tcaggcaggg aagttatagg cacaaagcta cgtctggaac
     2521 ccctttgtgc cccctttgtg ttcctcaagg aagcagtacc tttgaagaga tctctgctgc
     2581 attaagtgat gaccggctac gtttcatgtc aggcttgctt tgccttgtgg gctactcagt
     2641 gcagaacctg ctgtaaccct cagttcaaaa aatggactgg caatgtgatt agcgttggat
     2701 gctttaccat tctctttagc ttggaaagtc ctctccactg actgtagcct ccaattcact
     2761 ggccgtcgtt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     2821 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     2881 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatgcga tttattcaac
     2941 aaagccgccg tcccgtcaag tcagcgtaat gctctgccag tgttacaacc aattaaccaa
     3001 ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatatattca tatcaggatt
     3061 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     3121 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     3181 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     3241 gacgactgaa tccggtgaga atggcaaaag cttatgcatt tctttccaga cttgttcaac
     3301 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     3361 tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     3421 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     3481 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     3541 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     3601 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     3661 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     3721 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     3781 tcgcggcttc gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     3841 gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta
     3901 acatcagaga ttttgagaca caacgtggct ttgttgaata aatcgaactt ttgctgagtt
     3961 gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt tccgtggcaa agcaaaagtt
     4021 caaaatcacc aactggtcca cctacaacaa agctctcatc aaccgtggct ccctcacttt
     4081 ctggctggat gatggggcga ttcaggcctg gtatgagtca gcaacacctt cttcacgagg
     4141 cagacctctc gacggagttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4201 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4261 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4321 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4381 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4441 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4501 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4561 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4621 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4681 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4741 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4801 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4861 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4921 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gaagctcgca
     4981 cattcagcag cgtttttcag cgcgttttcg atcaacgttt caatgttggt atcaacacca
     5041 ggtttaactt tgaacttatc ggcactgacg gttactgatt ttgaactttt gctttgccac
     5101 ggaacggtct gcgttgtcgg gaagatgcgt gatctgatcc ttcaactcag caaaagttcc
     5161 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     5221 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     5281 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     5341 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cttgcatgcc
     5401 tgcaggtcga ctctagagga tccccgggta ccgagctcga attagcttgc atgcctgcag
     5461 gtcaattccc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     5521 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     5581 gcgtggatag cggtttgact cacggggatt tcaagtctcc accccattga cgtcaatggg
     5641 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     5701 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
     5761 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca taggagacac
     5821 cgggaccgat ccagcctggg gatcccgacc atggcttcgt acccctgcca tcaacacgcg
     5881 tctgcgttcg accaggctgc gcgttctcgc ggccatagca accgacgtac ggcgttgcgc
     5941 cctcgccggc agcaagaagc cacggaagtc cgcctggagc agaaaatgcc cacgctactg
     6001 cgggtttata tagacggtcc tcacgggatg gggaaaacca ccaccacgca actgctggtg
     6061 gccctgggtt cgcgcgacga tatcgtctac gtacccgagc cgatgactta ctggcaggtg
     6121 ctgggggctt ccgagacaat cgcgaacatc tacaccacac aacaccgcct cgaccagggt
     6181 gagatatcgg ccggggacgc ggcggtggta atgacaagcg cccagataac aatgggcatg
     6241 ccttatgccg tgaccgacgc cgttctggct cctcatatcg ggggggaggc tgggagctca
     6301 catgccccgc ccccggccct caccctcatc ttcgaccgcc atcccatcgc cgccctcctg
     6361 tgctacccgg ccgcgcgata ccttatgggc agcatgaccc cccaggccgt gctggcgttc
     6421 gtggccctca tcccgccgac cttgcccggc acaaacatcg tgttgggggc ccttccggag
     6481 gacagacaca tcgaccgcct ggccaaacgc cagcgccccg gcgagcggct tgacctggct
     6541 atgctggccg cgattcgccg cgtttacggg ctgcttgcca atacggtgcg gtatctgcag
     6601 ggcggcgggt cgtggcggga ggattgggga cagctttcgg ggacggccgt gccgccccag
     6661 ggtgccgagc cccagagcaa cgcgggccca cgaccccata tcggggacac gttatttacc
     6721 ctgtttcggg cccccgagtt gctggccccc aacggcgacc tgtacaacgt gtttgcctgg
     6781 gccttggacg tcttggccaa acgcctccgt cccatgcacg tctttatcct ggattacgac
     6841 caatcgcccg ccggctgccg ggacgccctg ctgcaactta cctccgggat ggtccagacc
     6901 cacgtcacca cccccggctc cataccgacg atctgcgacc tggcgcgcac gtttgcccgg
     6961 gagatggggg
//
