LOCUS       sequence-171020-        6970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human ZGPAT.
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_ZGPAT
FEATURES             Location/Qualifiers
     source          1..6970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      22..610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     511..530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     764..781
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1081..1102
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1117..1122,1123..1140,1141..1147)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1155..1171
                     /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     1160..1182
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1179..1195
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1179..1195
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1372..1574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1525..1545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1571..1595
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1671..2801
                     /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             3835..3858
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3862..3885
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3889..3912
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3928..3984
                     /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             3997..4791
                     /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(4051..4070)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4661..4680
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5562..5578)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5571..5593)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(5797..6612)
                     /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     6520..6539
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6604..6623
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
       61 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
      121 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
      181 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
      241 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
      301 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
      361 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
      421 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
      481 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
      541 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
      601 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      661 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
      721 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      781 ggaagcggaa gaagctcgca cattcagcag cgtttttcag cgcgttttcg atcaacgttt
      841 caatgttggt atcaacacca ggtttaactt tgaacttatc ggcactgacg gttactgatt
      901 ttgaactttt gctttgccac ggaacggtct gcgttgtcgg gaagatgcgt gatctgatcc
      961 ttcaactcag caaaagttcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     1021 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     1081 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     1141 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     1201 ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccgggta ccgagctcga
     1261 attagcttgc atgcctgcag gtcaattccc tggcattatg cccagtacat gaccttatgg
     1321 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     1381 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tcaagtctcc
     1441 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     1501 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     1561 atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt
     1621 ttgacctcca taggagacac cgggaccgat ccagcctggg gatcccgacc atggcttcgt
     1681 acccctgcca tcaacacgcg tctgcgttcg accaggctgc gcgttctcgc ggccatagca
     1741 accgacgtac ggcgttgcgc cctcgccggc agcaagaagc cacggaagtc cgcctggagc
     1801 agaaaatgcc cacgctactg cgggtttata tagacggtcc tcacgggatg gggaaaacca
     1861 ccaccacgca actgctggtg gccctgggtt cgcgcgacga tatcgtctac gtacccgagc
     1921 cgatgactta ctggcaggtg ctgggggctt ccgagacaat cgcgaacatc tacaccacac
     1981 aacaccgcct cgaccagggt gagatatcgg ccggggacgc ggcggtggta atgacaagcg
     2041 cccagataac aatgggcatg ccttatgccg tgaccgacgc cgttctggct cctcatatcg
     2101 ggggggaggc tgggagctca catgccccgc ccccggccct caccctcatc ttcgaccgcc
     2161 atcccatcgc cgccctcctg tgctacccgg ccgcgcgata ccttatgggc agcatgaccc
     2221 cccaggccgt gctggcgttc gtggccctca tcccgccgac cttgcccggc acaaacatcg
     2281 tgttgggggc ccttccggag gacagacaca tcgaccgcct ggccaaacgc cagcgccccg
     2341 gcgagcggct tgacctggct atgctggccg cgattcgccg cgtttacggg ctgcttgcca
     2401 atacggtgcg gtatctgcag ggcggcgggt cgtggcggga ggattgggga cagctttcgg
     2461 ggacggccgt gccgccccag ggtgccgagc cccagagcaa cgcgggccca cgaccccata
     2521 tcggggacac gttatttacc ctgtttcggg cccccgagtt gctggccccc aacggcgacc
     2581 tgtacaacgt gtttgcctgg gccttggacg tcttggccaa acgcctccgt cccatgcacg
     2641 tctttatcct ggattacgac caatcgcccg ccggctgccg ggacgccctg ctgcaactta
     2701 cctccgggat ggtccagacc cacgtcacca cccccggctc cataccgacg atctgcgacc
     2761 tggcgcgcac gtttgcccgg gagatggggg aggctaactg aggatcccac gtcactattg
     2821 tatactctat attatactct atgttatact ctgtaatcct actcaataaa cgtgtcacgc
     2881 ctgtgaaacc gtactaagtc tcccgtgtat tcttatcacc atcaggtgac atcctcgccc
     2941 aggctgtcaa tcatgccggt atcgattcca gtagcaccgg ccccacgctg acaacccact
     3001 cttgcagcgt tagcagcgcc cctcttaaca agccgacccc caccagcgtc gcggttacta
     3061 acactcctct ccccgacctg cagcccagct ttgcctcgag ggaagtcgct ggaccagtgt
     3121 gtggagaccc tgcagaagca gaccagggtt ggcaaggctg gcaccaacaa gccccccagg
     3181 tgccggggaa gaggggccag gcctgggggc cgcccagctc ctcggaatgt gtttgacttc
     3241 ctcaatgaaa agctgcaagg tcaggctcct ggggccctag aagccggggc ggccccagcg
     3301 gggaggagga gcaaggacat gtaccatgcc agcaagagtg ccaagcgggc cctgagcctg
     3361 cggctcttcc agactgagga gaagatcgag cgaacccagc gggacatcag gagcatccag
     3421 gaggctctcg cccgcaacgc tggccggtac gtgtggggcc cagctcaggg caaagggcga
     3481 cccagggtgg ccctgccccc gggatacatg ctggaggtca cctgacccaa gcatagcggg
     3541 ctgagtccag gaggggtctg catgttggag gacgggcagc gggcacacag gcagtgggta
     3601 cggcagactg gggttggtgg catggcccag gaccccacgc tgactagtga gcccctccgc
     3661 aggcatagcg tggcgtcagc ccagctgcag gagaagctgg caggagccca gcgccagctg
     3721 gggcagctcc gggctcagga agccggcctg cagcaggagc agaggaaggc agacacccac
     3781 aagaagatga ctgagttcgg gagcggagga ggttccggtg gaggtggttc tggagattac
     3841 aaggatgacg acgataaggg cgattacaag gatgacgacg ataagggaga ttacaaggat
     3901 gacgacgata aggtttcagg aagcggagct actaacttca gcctgctgaa gcaggctgga
     3961 gacgtggagg agaaccctgg acctggatcg tttcgcatga ttgaacaaga tggattgcac
     4021 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4081 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4141 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4201 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4261 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4321 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4381 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4441 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4501 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     4561 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4621 tgtggccggc tgggtgtggc cgaccgctat caggacatag cgttggctac ccgtgatatt
     4681 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4741 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg attcgaacat
     4801 cctatggacg aagcgtggga ccccagcacg ggctgccctc aggaagacca gtgttgcccg
     4861 aggaggggcc ggcctgctgg cctggggcgt gcagacactg ctgagtggag acagagctgc
     4921 ggggtcccat ctggacactt acttgcccac ctgccagtgt cttgggcatt tccttggcaa
     4981 ggacattaaa gtgatttcat cacagtgtca ttcagtggag atcagctggc tgcagggtct
     5041 ctggggagag aagggtctgt gcagggctgt ggtgagagac gggctgggct ggaggggcct
     5101 aggaggccgg cattctccct tcttaccctt ctctgaccaa ggcagatcgg aggccagggt
     5161 gcccgccagg ctgcctgaca cccacagatg ggtggagctc agtgctgctc cagcgggcga
     5221 gacgggcggg agaggactgt ggcccaccac gtggggcact cttggtgagg caggcggacc
     5281 tggccctgtg ggtttggggg ttctgtgcca gctctgctca cgtcttcgtg tgtggagggg
     5341 ttgtgagtta ggctgtcaga acagtgcggg ctagaggcgc acacgtttca tctaggcttc
     5401 gggcggggcg cagcaggtgg gtgaggctgc ggtggtggct gctgcccagg gtgggaggct
     5461 caggggaggc gcttggcgac aggagcagga agtgatgagg tggggagctc ttggaaagtc
     5521 ctctccactg actgtagcct ccaattcact ggccgtcgtt cactggccgt cgttttacaa
     5581 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     5641 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     5701 agcctgaatg gcgaatgcga tttattcaac aaagccgccg tcccgtcaag tcagcgtaat
     5761 gctctgccag tgttacaacc aattaaccaa ttctgattag aaaaactcat cgagcatcaa
     5821 atgaaactgc aatatattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     5881 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     5941 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     6001 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     6061 cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     6121 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     6181 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     6241 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     6301 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     6361 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     6421 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     6481 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     6541 atataaatca gcatccatgt tggaatttaa tcgcggcttc gagcaagacg tttcccgttg
     6601 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     6661 tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtggct
     6721 ttgttgaata aatcgaactt ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa
     6781 cgcagaccgt tccgtggcaa agcaaaagtt caaaatcacc aactggtcca cctacaacaa
     6841 agctctcatc aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggcctg
     6901 gtatgagtca gcaacacctt cttcacgagg cagacctctc gacggagttc cactgagcgt
     6961 cagaccccgt
//
