LOCUS       sequence-271109-        6734 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human ZNF331.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6734)
  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 6734)
  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_ZNF331
FEATURES             Location/Qualifiers
     source          1..6734
                     /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"
     CDS             2228..2251
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2255..2278
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2282..2305
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2321..2377
                     /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             2390..3184
                     /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(2444..2463)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3054..3073
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3955..3971)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3964..3986)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(4190..5005)
                     /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     4913..4932
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4997..5016
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5385..5973
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5874..5893
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6127..6144
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6444..6465
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6480..6485,6486..6503,6504..6510)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6518..6534
                     /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     6523..6545
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6542..6558
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6542..6558
                     /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 agcccagctg gcctttactc gagtcaatta
     1741 ccttactcag catcagaaga tccacaccgg tgagaagcct cacgaatgta aggagtgtgg
     1801 gaaggccttt cgctggggtt cgagcctcgt taagcacgag aggatacata cgggcgagaa
     1861 gccgtacaag tgcacagaat gtgggaaggc cttcaattgt ggctatcacc tcactcagca
     1921 cgagagaatc cacacaggcg aaaccccgta taaatgtaag gagtgtggga aggctttcat
     1981 ttatggatcg agcctcgtga aacatgagag aattcatacc ggggtgaaac cctatgggtg
     2041 tacagaatgt gggaagagct ttagtcacgg ccatcagctt acacaacatc agaaaacgca
     2101 cagtggggcg aaatcctacg aatgtaagga gtgcgggaag gcatgtaacc acctaaacca
     2161 tctccgagaa catcagagga tccacaatag cgggagcgga ggaggttccg gtggaggtgg
     2221 ttctggagat tacaaggatg acgacgataa gggcgattac aaggatgacg acgataaggg
     2281 agattacaag gatgacgacg ataaggtttc aggaagcgga gctactaact tcagcctgct
     2341 gaagcaggct ggagacgtgg aggagaaccc tggacctgga tcgtttcgca tgattgaaca
     2401 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     2461 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     2521 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     2581 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     2641 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     2701 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     2761 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     2821 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     2881 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     2941 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     3001 tggattcatc gactgtggcc ggctgggtgt ggccgaccgc tatcaggaca tagcgttggc
     3061 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     3121 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     3181 ctgattcgaa catcgaacgc agtagcccgc tcgtatctat ggtttcgctt tccacagttt
     3241 gttacctgca gtcaactgca gttcaaaaat attaaatgga aaattccaga aataaagaat
     3301 tttaagtctc aaatggtgtg cccttctgag tagcgtgatg aaatctctcg ctgtccggct
     3361 ccagccggcc ggggatgtga gtcatccctt ggtccagcac atccacgctg tatacgccac
     3421 ccaccctgct agtgacttag tagccgtctt ggtgatcaga tcaactatcc cagcatcaca
     3481 gtgcctgtgc ccaagcagtc ctcactttgc ttaacagtgg ccccagagag caggagtagt
     3541 gatgctggtg attcggatat gccaaagaga agccacaaag tgcttccttt taaatgaaaa
     3601 ggtgaaagtt ctcaacttaa aaaagaaaag aaaaaaatca tatactgagg ttgctaagat
     3661 ctacatgaca ataagatatt ttgggagaga gatacgttca cataatttta ttacatatat
     3721 tgttacaatt gttctatttt gttagttact gttggtaatc tcttactgtg cctaatttat
     3781 aaattaaact ttagatatgt atgtacagga aaaaatgtag tatatagtat cgtatatatg
     3841 tacaggaaaa aaacgtacta tctgtggttt caggtgtcca ctgcgggtct tcaaacgtac
     3901 cccttggaaa gtcctctcca ctgactgtag cctccaattc actggccgtc gttcactggc
     3961 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     4021 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     4081 ccaacagttg cgcagcctga atggcgaatg cgatttattc aacaaagccg ccgtcccgtc
     4141 aagtcagcgt aatgctctgc cagtgttaca accaattaac caattctgat tagaaaaact
     4201 catcgagcat caaatgaaac tgcaatatat tcatatcagg attatcaata ccatattttt
     4261 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     4321 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     4381 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     4441 agaatggcaa aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     4501 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     4561 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     4621 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     4681 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     4741 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     4801 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     4861 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     4921 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ttcgagcaag
     4981 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
     5041 gttttattgt tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag
     5101 acacaacgtg gctttgttga ataaatcgaa cttttgctga gttgaaggat cagatcacgc
     5161 atcttcccga caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc accaactggt
     5221 ccacctacaa caaagctctc atcaaccgtg gctccctcac tttctggctg gatgatgggg
     5281 cgattcaggc ctggtatgag tcagcaacac cttcttcacg aggcagacct ctcgacggag
     5341 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5401 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5461 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5521 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5581 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5641 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5701 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5761 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5821 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5881 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5941 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6001 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6061 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6121 gagcgcagcg agtcagtgag cgaggaagcg gaagaagctc gcacattcag cagcgttttt
     6181 cagcgcgttt tcgatcaacg tttcaatgtt ggtatcaaca ccaggtttaa ctttgaactt
     6241 atcggcactg acggttactg attttgaact tttgctttgc cacggaacgg tctgcgttgt
     6301 cgggaagatg cgtgatctga tccttcaact cagcaaaagt tcccaatacg caaaccgcct
     6361 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     6421 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     6481 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     6541 acaggaaaca gctatgacca tgattacgcc aagcttgcat gcctgcaggt cgactctaga
     6601 ggatccccgg gtaccgagct cgaattagct tgcatgcctg caggtcaatt ccctggcatt
     6661 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     6721 tcgctattac catg
//
