LOCUS       sequence-168159-        6965 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human ZNF580.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6965)
  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 6965)
  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_ZNF580
FEATURES             Location/Qualifiers
     source          1..6965
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             740..763
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             767..790
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             794..817
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             833..889
                     /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             902..1696
                     /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(956..975)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1566..1585
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2467..2483)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2476..2498)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2702..3517)
                     /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     3425..3444
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3509..3528
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3897..4485
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4386..4405
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4639..4656
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4956..4977
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4992..4997,4998..5015,5016..5022)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5030..5046
                     /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     5035..5057
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5054..5070
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5054..5070
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5247..5449
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5400..5420
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5446..5470
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5546..6676
                     /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 agcttgatgg aacgtgggag agccgggctg gagtcacagt ttttattttg ctagggaggt
       61 agtcagtggc gcacaaaggg taacaagcag tgatagtggg gatgctttcc attttaggaa
      121 acttttccta aaaggaagtg gcaattttaa ctaatctccc ctccgccgct cccagctgcc
      181 gctccagatg ctgctgctgc cgccgcggcc accccaccct cggtcctcct ctccggaggc
      241 catggaccca ccgcccccca aggctccccc tttccccaag gcggaaggcc cctcctccac
      301 tccttcctcg gcggcggggc cccgaccccc gcggctgggc cgccacctcc tcatcgacgc
      361 caatggggtc ccctacacat acacggtgca gctggaggag gagccccggg gcccgcccca
      421 gcgcgaggcg cccccaggag agcccggccc tcgcaagggc tacagctgcc cggagtgcgc
      481 ccgtgtcttt gccagccctc tgcggctgca gagccaccgc gtgtcgcact cggacctcaa
      541 gcccttcacg tgcggcgcct gcggcaaggc cttcaagcgc tccagccacc tgtcgcggca
      601 tcgcgccacg caccgcgccc gcgccgggcc gccgcacacc tgcccgctct gcccacgccg
      661 cttccaggac gccgcggagc tggcgcagca cgtgcgcctc catgggagcg gaggaggttc
      721 cggtggaggt ggttctggag attacaagga tgacgacgat aagggcgatt acaaggatga
      781 cgacgataag ggagattaca aggatgacga cgataaggtt tcaggaagcg gagctactaa
      841 cttcagcctg ctgaagcagg ctggagacgt ggaggagaac cctggacctg gatcgtttcg
      901 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
      961 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     1021 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     1081 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     1141 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     1201 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     1261 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     1321 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     1381 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     1441 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     1501 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggccgacc gctatcagga
     1561 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     1621 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     1681 tgacgagttc ttctgattcg aacatcaccc ggcctgtgct gccctgcccg tctcagggcc
     1741 accaagtctg acccacacag cgtcactcac tcccacacac accccctggc tctgctgagg
     1801 ttactgcctt accctgggcc tcagtttccc caccttccaa agggaggagc atcattcctt
     1861 ccttaccccc tttctagctg tgtgatgtag accaaagtcg ttgcccctcc ctgggcctgg
     1921 gaaccagtcg gaactgggtt ccagtccagc tgtgctgtgt gagcctgtgc aagtgacatg
     1981 acctctctaa accttggttt tctgctctct ggagcggtga accggtggtt gtctgcgggg
     2041 aagagatgat aaagagcacg ggcacggtct ggttcatttc tgtatctacc ccccttccgc
     2101 ccacgccccc gaccctttgc tcaataaaca ttccgcactc cattttcagg ctcttccttg
     2161 cgtgtgcgtg cacgttgggt gctggggggt ggagaccgga tcttcctcgc ttgggtactt
     2221 tcctctcggt gtgtgtttct ggccggagcc gtttcgcgac ggcccgggcg ccccgcccca
     2281 accttccttc cctagaccct cttctctccc ttcggcttct ctctttcggc cggcgccgcc
     2341 agttcctggg gcacacccag aggtcccctt ctcgccgccg cctgcaactg cgagggtagc
     2401 ccggggccgc ttggcttgga aagtcctctc cactgactgt agcctccaat tcactggccg
     2461 tcgttcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     2521 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     2581 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgcgatttat tcaacaaagc
     2641 cgccgtcccg tcaagtcagc gtaatgctct gccagtgtta caaccaatta accaattctg
     2701 attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     2761 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     2821 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     2881 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     2941 ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt tcaacaggcc
     3001 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     3061 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
     3121 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     3181 attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat
     3241 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     3301 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     3361 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga
     3421 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     3481 gcttcgagca agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta
     3541 tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc
     3601 agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct gagttgaagg
     3661 atcagatcac gcatcttccc gacaacgcag accgttccgt ggcaaagcaa aagttcaaaa
     3721 tcaccaactg gtccacctac aacaaagctc tcatcaaccg tggctccctc actttctggc
     3781 tggatgatgg ggcgattcag gcctggtatg agtcagcaac accttcttca cgaggcagac
     3841 ctctcgacgg agttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3901 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3961 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4021 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4081 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4141 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4201 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4261 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4321 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4381 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4441 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4501 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4561 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4621 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagaagc tcgcacattc
     4681 agcagcgttt ttcagcgcgt tttcgatcaa cgtttcaatg ttggtatcaa caccaggttt
     4741 aactttgaac ttatcggcac tgacggttac tgattttgaa cttttgcttt gccacggaac
     4801 ggtctgcgtt gtcgggaaga tgcgtgatct gatccttcaa ctcagcaaaa gttcccaata
     4861 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     4921 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     4981 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     5041 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc atgcctgcag
     5101 gtcgactcta gaggatcccc gggtaccgag ctcgaattag cttgcatgcc tgcaggtcaa
     5161 ttccctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     5221 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     5281 gatagcggtt tgactcacgg ggatttcaag tctccacccc attgacgtca atgggagttt
     5341 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     5401 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
     5461 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagga gacaccggga
     5521 ccgatccagc ctggggatcc cgaccatggc ttcgtacccc tgccatcaac acgcgtctgc
     5581 gttcgaccag gctgcgcgtt ctcgcggcca tagcaaccga cgtacggcgt tgcgccctcg
     5641 ccggcagcaa gaagccacgg aagtccgcct ggagcagaaa atgcccacgc tactgcgggt
     5701 ttatatagac ggtcctcacg ggatggggaa aaccaccacc acgcaactgc tggtggccct
     5761 gggttcgcgc gacgatatcg tctacgtacc cgagccgatg acttactggc aggtgctggg
     5821 ggcttccgag acaatcgcga acatctacac cacacaacac cgcctcgacc agggtgagat
     5881 atcggccggg gacgcggcgg tggtaatgac aagcgcccag ataacaatgg gcatgcctta
     5941 tgccgtgacc gacgccgttc tggctcctca tatcgggggg gaggctggga gctcacatgc
     6001 cccgcccccg gccctcaccc tcatcttcga ccgccatccc atcgccgccc tcctgtgcta
     6061 cccggccgcg cgatacctta tgggcagcat gaccccccag gccgtgctgg cgttcgtggc
     6121 cctcatcccg ccgaccttgc ccggcacaaa catcgtgttg ggggcccttc cggaggacag
     6181 acacatcgac cgcctggcca aacgccagcg ccccggcgag cggcttgacc tggctatgct
     6241 ggccgcgatt cgccgcgttt acgggctgct tgccaatacg gtgcggtatc tgcagggcgg
     6301 cgggtcgtgg cgggaggatt ggggacagct ttcggggacg gccgtgccgc cccagggtgc
     6361 cgagccccag agcaacgcgg gcccacgacc ccatatcggg gacacgttat ttaccctgtt
     6421 tcgggccccc gagttgctgg cccccaacgg cgacctgtac aacgtgtttg cctgggcctt
     6481 ggacgtcttg gccaaacgcc tccgtcccat gcacgtcttt atcctggatt acgaccaatc
     6541 gcccgccggc tgccgggacg ccctgctgca acttacctcc gggatggtcc agacccacgt
     6601 caccaccccc ggctccatac cgacgatctg cgacctggcg cgcacgtttg cccgggagat
     6661 gggggaggct aactgaggat cccacgtcac tattgtatac tctatattat actctatgtt
     6721 atactctgta atcctactca ataaacgtgt cacgcctgtg aaaccgtact aagtctcccg
     6781 tgtattctta tcaccatcag gtgacatcct cgcccaggct gtcaatcatg ccggtatcga
     6841 ttccagtagc accggcccca cgctgacaac ccactcttgc agcgttagca gcgcccctct
     6901 taacaagccg acccccacca gcgtcgcggt tactaacact cctctccccg acctgcagcc
     6961 cagct
//
