LOCUS       sequence-190326-        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             737..760
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             764..787
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             791..814
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             830..886
                     /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             899..1693
                     /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(953..972)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1563..1582
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2464..2480)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2473..2495)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2699..3514)
                     /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     3422..3441
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3506..3525
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3894..4482
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4383..4402
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4636..4653
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4953..4974
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4989..4994,4995..5012,5013..5019)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5027..5043
                     /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     5032..5054
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5051..5067
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5051..5067
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5244..5446
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5397..5417
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5443..5467
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5543..6673
                     /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 ttgatggaac gtgggagagc cgggctggag tcacagtttt tattttgcta gggaggtagt
       61 cagtggcgca caaagggtaa caagcagtga tagtggggat gctttccatt ttaggaaact
      121 tttcctaaaa ggaagtggca attttaacta atctcccctc cgccgctccc agctgccgct
      181 ccagatgctg ctgctgccgc cgcggccacc ccaccctcgg tcctcctctc cggaggccat
      241 ggacccaccg ccccccaagg ctcccccttt ccccaaggcg gaaggcccct cctccactcc
      301 ttcctcggcg gcggggcccc gacccccgcg gctgggccgc cacctcctca tcgacgccaa
      361 tggggtcccc tacacataca cggtgcagct ggaggaggag ccccggggcc cgccccagcg
      421 cgaggcgccc ccaggagagc ccggccctcg caagggctac agctgcccgg agtgcgcccg
      481 tgtctttgcc agccctctgc ggctgcagag ccaccgcgtg tcgcactcgg acctcaagcc
      541 cttcacgtgc ggcgcctgcg gcaaggcctt caagcgctcc agccacctgt cgcggcatcg
      601 cgccacgcac cgcgcccgcg ccgggccgcc gcacacctgc ccgctctgcc cacgccgctt
      661 ccaggacgcc gcggagctgg cgcagcacgt gcgcctccat gggagcggag gaggttccgg
      721 tggaggtggt tctggagatt acaaggatga cgacgataag ggcgattaca aggatgacga
      781 cgataaggga gattacaagg atgacgacga taaggtttca ggaagcggag ctactaactt
      841 cagcctgctg aagcaggctg gagacgtgga ggagaaccct ggacctggat cgtttcgcat
      901 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
      961 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     1021 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     1081 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     1141 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     1201 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     1261 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     1321 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     1381 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     1441 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     1501 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gccgaccgct atcaggacat
     1561 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     1621 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     1681 cgagttcttc tgattcgaac atcacccggc ctgtgctgcc ctgcccgtct cagggccacc
     1741 aagtctgacc cacacagcgt cactcactcc cacacacacc ccctggctct gctgaggtta
     1801 ctgccttacc ctgggcctca gtttccccac cttccaaagg gaggagcatc attccttcct
     1861 tacccccttt ctagctgtgt gatgtagacc aaagtcgttg cccctccctg ggcctgggaa
     1921 ccagtcggaa ctgggttcca gtccagctgt gctgtgtgag cctgtgcaag tgacatgacc
     1981 tctctaaacc ttggttttct gctctctgga gcggtgaacc ggtggttgtc tgcggggaag
     2041 agatgataaa gagcacgggc acggtctggt tcatttctgt atctaccccc cttccgccca
     2101 cgcccccgac cctttgctca ataaacattc cgcactccat tttcaggctc ttccttgcgt
     2161 gtgcgtgcac gttgggtgct ggggggtgga gaccggatct tcctcgcttg ggtactttcc
     2221 tctcggtgtg tgtttctggc cggagccgtt tcgcgacggc ccgggcgccc cgccccaacc
     2281 ttccttccct agaccctctt ctctcccttc ggcttctctc tttcggccgg cgccgccagt
     2341 tcctggggca cacccagagg tccccttctc gccgccgcct gcaactgcga gggtagcccg
     2401 gggccgcttg gcttggaaag tcctctccac tgactgtagc ctccaattca ctggccgtcg
     2461 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     2521 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     2581 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgc gatttattca acaaagccgc
     2641 cgtcccgtca agtcagcgta atgctctgcc agtgttacaa ccaattaacc aattctgatt
     2701 agaaaaactc atcgagcatc aaatgaaact gcaatatatt catatcagga ttatcaatac
     2761 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
     2821 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     2881 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
     2941 aatccggtga gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc
     3001 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
     3061 cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
     3121 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     3181 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat
     3241 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
     3301 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     3361 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat
     3421 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggct
     3481 tcgagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
     3541 aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga
     3601 gattttgaga cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc
     3661 agatcacgca tcttcccgac aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca
     3721 ccaactggtc cacctacaac aaagctctca tcaaccgtgg ctccctcact ttctggctgg
     3781 atgatggggc gattcaggcc tggtatgagt cagcaacacc ttcttcacga ggcagacctc
     3841 tcgacggagt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3901 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3961 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4021 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4081 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4141 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4201 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4261 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4321 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4381 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4441 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4501 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4561 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4621 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagaagctcg cacattcagc
     4681 agcgtttttc agcgcgtttt cgatcaacgt ttcaatgttg gtatcaacac caggtttaac
     4741 tttgaactta tcggcactga cggttactga ttttgaactt ttgctttgcc acggaacggt
     4801 ctgcgttgtc gggaagatgc gtgatctgat ccttcaactc agcaaaagtt cccaatacgc
     4861 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     4921 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     4981 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     5041 caatttcaca caggaaacag ctatgaccat gattacgcca agcttgcatg cctgcaggtc
     5101 gactctagag gatccccggg taccgagctc gaattagctt gcatgcctgc aggtcaattc
     5161 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     5221 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     5281 agcggtttga ctcacgggga tttcaagtct ccaccccatt gacgtcaatg ggagtttgtt
     5341 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     5401 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg
     5461 tcagatcgcc tggagacgcc atccacgctg ttttgacctc cataggagac accgggaccg
     5521 atccagcctg gggatcccga ccatggcttc gtacccctgc catcaacacg cgtctgcgtt
     5581 cgaccaggct gcgcgttctc gcggccatag caaccgacgt acggcgttgc gccctcgccg
     5641 gcagcaagaa gccacggaag tccgcctgga gcagaaaatg cccacgctac tgcgggttta
     5701 tatagacggt cctcacggga tggggaaaac caccaccacg caactgctgg tggccctggg
     5761 ttcgcgcgac gatatcgtct acgtacccga gccgatgact tactggcagg tgctgggggc
     5821 ttccgagaca atcgcgaaca tctacaccac acaacaccgc ctcgaccagg gtgagatatc
     5881 ggccggggac gcggcggtgg taatgacaag cgcccagata acaatgggca tgccttatgc
     5941 cgtgaccgac gccgttctgg ctcctcatat cgggggggag gctgggagct cacatgcccc
     6001 gcccccggcc ctcaccctca tcttcgaccg ccatcccatc gccgccctcc tgtgctaccc
     6061 ggccgcgcga taccttatgg gcagcatgac cccccaggcc gtgctggcgt tcgtggccct
     6121 catcccgccg accttgcccg gcacaaacat cgtgttgggg gcccttccgg aggacagaca
     6181 catcgaccgc ctggccaaac gccagcgccc cggcgagcgg cttgacctgg ctatgctggc
     6241 cgcgattcgc cgcgtttacg ggctgcttgc caatacggtg cggtatctgc agggcggcgg
     6301 gtcgtggcgg gaggattggg gacagctttc ggggacggcc gtgccgcccc agggtgccga
     6361 gccccagagc aacgcgggcc cacgacccca tatcggggac acgttattta ccctgtttcg
     6421 ggcccccgag ttgctggccc ccaacggcga cctgtacaac gtgtttgcct gggccttgga
     6481 cgtcttggcc aaacgcctcc gtcccatgca cgtctttatc ctggattacg accaatcgcc
     6541 cgccggctgc cgggacgccc tgctgcaact tacctccggg atggtccaga cccacgtcac
     6601 cacccccggc tccataccga cgatctgcga cctggcgcgc acgtttgccc gggagatggg
     6661 ggaggctaac tgaggatccc acgtcactat tgtatactct atattatact ctatgttata
     6721 ctctgtaatc ctactcaata aacgtgtcac gcctgtgaaa ccgtactaag tctcccgtgt
     6781 attcttatca ccatcaggtg acatcctcgc ccaggctgtc aatcatgccg gtatcgattc
     6841 cagtagcacc ggccccacgc tgacaaccca ctcttgcagc gttagcagcg cccctcttaa
     6901 caagccgacc cccaccagcg tcgcggttac taacactcct ctccccgacc tgcagcccag
     6961 ctagc
//
