LOCUS       sequence-271146-        6694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human ZNF503.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6694)
  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 6694)
  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_ZNF503
FEATURES             Location/Qualifiers
     source          1..6694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..203
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     154..174
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     200..224
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             300..1430
                     /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"
     regulatory      1912..1921
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2188..2211
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2215..2238
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2242..2265
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2281..2337
                     /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             2350..3144
                     /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(2404..2423)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3014..3033
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3915..3931)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3924..3946)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(4150..4965)
                     /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     4873..4892
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4957..4976
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5345..5933
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5834..5853
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6087..6104
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6404..6425
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6440..6445,6446..6463,6464..6470)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6478..6494
                     /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     6483..6505
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6502..6518
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6502..6518
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      181 gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
      241 cacgctgttt tgacctccat aggagacacc gggaccgatc cagcctgggg atcccgacca
      301 tggcttcgta cccctgccat caacacgcgt ctgcgttcga ccaggctgcg cgttctcgcg
      361 gccatagcaa ccgacgtacg gcgttgcgcc ctcgccggca gcaagaagcc acggaagtcc
      421 gcctggagca gaaaatgccc acgctactgc gggtttatat agacggtcct cacgggatgg
      481 ggaaaaccac caccacgcaa ctgctggtgg ccctgggttc gcgcgacgat atcgtctacg
      541 tacccgagcc gatgacttac tggcaggtgc tgggggcttc cgagacaatc gcgaacatct
      601 acaccacaca acaccgcctc gaccagggtg agatatcggc cggggacgcg gcggtggtaa
      661 tgacaagcgc ccagataaca atgggcatgc cttatgccgt gaccgacgcc gttctggctc
      721 ctcatatcgg gggggaggct gggagctcac atgccccgcc cccggccctc accctcatct
      781 tcgaccgcca tcccatcgcc gccctcctgt gctacccggc cgcgcgatac cttatgggca
      841 gcatgacccc ccaggccgtg ctggcgttcg tggccctcat cccgccgacc ttgcccggca
      901 caaacatcgt gttgggggcc cttccggagg acagacacat cgaccgcctg gccaaacgcc
      961 agcgccccgg cgagcggctt gacctggcta tgctggccgc gattcgccgc gtttacgggc
     1021 tgcttgccaa tacggtgcgg tatctgcagg gcggcgggtc gtggcgggag gattggggac
     1081 agctttcggg gacggccgtg ccgccccagg gtgccgagcc ccagagcaac gcgggcccac
     1141 gaccccatat cggggacacg ttatttaccc tgtttcgggc ccccgagttg ctggccccca
     1201 acggcgacct gtacaacgtg tttgcctggg ccttggacgt cttggccaaa cgcctccgtc
     1261 ccatgcacgt ctttatcctg gattacgacc aatcgcccgc cggctgccgg gacgccctgc
     1321 tgcaacttac ctccgggatg gtccagaccc acgtcaccac ccccggctcc ataccgacga
     1381 tctgcgacct ggcgcgcacg tttgcccggg agatggggga ggctaactga ggatcccacg
     1441 tcactattgt atactctata ttatactcta tgttatactc tgtaatccta ctcaataaac
     1501 gtgtcacgcc tgtgaaaccg tactaagtct cccgtgtatt cttatcacca tcaggtgaca
     1561 tcctcgccca ggctgtcaat catgccggta tcgattccag tagcaccggc cccacgctga
     1621 caacccactc ttgcagcgtt agcagcgccc ctcttaacaa gccgaccccc accagcgtcg
     1681 cggttactaa cactcctctc cccgacctgc agcccagctt acggctttat gctccctaac
     1741 gacccactcc cccacatctg caactgggtg tcggccaacg ggccgtgcga caagcgcttc
     1801 gccacgtccg aagagctgct gagccacttg cggacccata cggcatttcc cgggacagac
     1861 aaactgctgt cgggctaccc cagctcgtcg tctctggcca gcgctgccgc ggccgccatg
     1921 gcttgccaca tgcacatccc cacctcgggc gcaccgggca gccctgggac gctggcgctg
     1981 cgcagccccc accacgcgct gggactcagc agccgctacc acccctactc caagagcccg
     2041 cttcccacgc ctggcgcccc cgtgccggtg cccgccgcca ccggaccgta ctactccccc
     2101 tacgccctct acggacagag actgaccacc gcctcggcgc tggggtatca agggagcgga
     2161 ggaggttccg gtggaggtgg ttctggagat tacaaggatg acgacgataa gggcgattac
     2221 aaggatgacg acgataaggg agattacaag gatgacgacg ataaggtttc aggaagcgga
     2281 gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctgga
     2341 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     2401 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     2461 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     2521 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     2581 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     2641 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     2701 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     2761 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     2821 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     2881 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     2941 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggccgaccgc
     3001 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3061 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     3121 cgccttcttg acgagttctt ctgattcgaa catcgggcgg ccgggagggc gagcgaggga
     3181 gaggagggag agggggaggg gaggagtcca gggagaggcg ggatcacggc ccaggctgct
     3241 gacacccgcg cgtggggagg actcgggcca cgaaaggaaa gaaatgtata ccgtatctat
     3301 ctacccgaca gcagcgaccg agacccggtg ggacactccc cttctcccca ctttcacctc
     3361 cccacccaaa ctttataaaa gttgaaaaaa tatcatttga ctttttatag aaaaaaaaag
     3421 gaaaaaataa ttgagaaagt gttcatctga ggactgcatc ggtggacact ggtatttatt
     3481 tatgttagct ccaagcggac cggtggttca aaagtgcatt atttagtttg agctctgtag
     3541 gtaaaaagga ggtgggaaaa attttaaaac ttgagggtaa aaatgtggaa aacaaaccct
     3601 cccatccctt gtagattata aataaaagca aaaccgccac agaactagag gtcttctctt
     3661 taatgttact ttaaaattgc tatgattgta ttgtacgtta tttaatgtct gattgaaaca
     3721 caaatttaca tgcatgtttg ttacaaaaaa aatgaaaaaa aaagtcacaa tttgtcagct
     3781 ctgatttcaa attgcaatta tttttaaggt gtataccatc gaagagaatg ggtatttttt
     3841 tgtatgtatt ctggaagaaa accttggaaa gtcctctcca ctgcctgtag cctccaattc
     3901 actggccgtc gttcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     3961 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     4021 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg cgatttattc
     4081 aacaaagccg ccgtcccgtc aagtcagcgt aatgctctgc cagtgttaca accaattaac
     4141 caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatatat tcatatcagg
     4201 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     4261 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     4321 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     4381 agtgacgact gaatccggtg agaatggcaa aagcttatgc atttctttcc agacttgttc
     4441 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     4501 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     4561 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     4621 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     4681 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     4741 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     4801 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     4861 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     4921 taatcgcggc ttcgagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     4981 actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat cttgtgcaat
     5041 gtaacatcag agattttgag acacaacgtg gctttgttga ataaatcgaa cttttgctga
     5101 gttgaaggat cagatcacgc atcttcccga caacgcagac cgttccgtgg caaagcaaaa
     5161 gttcaaaatc accaactggt ccacctacaa caaagctctc atcaaccgtg gctccctcac
     5221 tttctggctg gatgatgggg cgattcaggc ctggtatgag tcagcaacac cttcttcacg
     5281 aggcagacct ctcgacggag ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5341 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5401 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5461 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5521 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5581 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5641 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5701 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5761 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5821 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5881 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5941 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6001 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     6061 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagaagctc
     6121 gcacattcag cagcgttttt cagcgcgttt tcgatcaacg tttcaatgtt ggtatcaaca
     6181 ccaggtttaa ctttgaactt atcggcactg acggttactg attttgaact tttgctttgc
     6241 cacggaacgg tctgcgttgt cgggaagatg cgtgatctga tccttcaact cagcaaaagt
     6301 tcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     6361 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     6421 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     6481 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcttgcat
     6541 gcctgcaggt cgactctaga ggatccccgg gtaccgagct cgaattagct tgcatgcctg
     6601 caggtcaatt ccctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     6661 gtacatctac gtattagtca tcgctattac catg
//
