LOCUS       sequence-191465-        4268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor Vector containing ZNF66 transcription factor, part of the 
            Human TFome Collection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4268)
  AUTHORS   Ng AHM, Khoshakhlagh P, Rojo Arias JE, Pasquini G, Wang K, Swiersy 
            A, Shipman SL, Appleton E, Kiaee K, Kohman RE, Vernet A, Dysart M, 
            Leeper K, Saylor W, Huang JY, Graveline A, Taipale J, Hill DE, Vidal
            M, Melero-Martin JM, Busskamp V, Church GM
  TITLE     A comprehensive library of human transcription factors for cell fate
            engineering.
  JOURNAL   Nat Biotechnol. 2021 Apr;39(4):510-519. doi: 
            10.1038/s41587-020-0742-6. Epub 2020 Nov 30.
  PUBMED    33257861
REFERENCE   2  (bases 1 to 4268)
  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"; doi: 
            "10.1038/s41587-020-0742-6"; journalName: "Nat Biotechnol"; date: 
            "2021-04"; volume: "39"; issue: "4"; pages: "510-519"
COMMENT     Sequence Label: pDONR221-ZNF66
FEATURES             Location/Qualifiers
     source          1..4268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1723..1822)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1839..1858)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1840..1858)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1863..1879)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1863..1879)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1981..2000)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1992..2801
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2059..2078)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2948..3536
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3437..3456
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3690..3707
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3866..3893)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3985..4071)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4120..4142
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4134..4151
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4135..4151
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4167..4266
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 atgggccccc tgcagttcag agatgtggcc atcgagttca gcctggagga gtggcactgc
       61 ctggacatgg cccagaggaa cctgtacaga gacgtgatgc tggagaacta cagaaacctg
      121 gtgttcctgg gcatcgtggt gagcaagccc gacctgatca cccacctgga gcagggcaag
      181 aagcctagca ccatgcagag acacgagatg gtggccaacc ctagcgtgct gtgcagccac
      241 ttcaaccagg acctgtggcc cgagcagagc atcaaggaca gcttccagaa gctgatcctg
      301 agaagacaca agaagtgcgg ccacgacaac ctgcagctga agaagggctg cgagagcgtg
      361 gacaagtgca aggtgcacaa gagaggctac aacggcctga accagtgcct gacaaccacc
      421 cagagcaaga tgttccagtg cgacaagcac ggcaaggtgt tccaccagtt cagcaacacc
      481 aacaggcaca agattagaca cactggcaag aacccctgca agttcaccga atgtggcaaa
      541 gcttttaaca ggagctccac cttcacaacc cacaaaaaga tccatacagg cgagaaacct
      601 tataagtgca tcgagtgtgg caaggcattc aacagatcct cccacctgac cacccacaag
      661 ataatccata ccggcgaaaa aagatacaag tgtgaagact gcggtaaggc atttaaccgg
      721 tctagcaacc tcaccaccca taagaagata cataccggtg agaagccata caagtgtgaa
      781 gaatgtggga aggcgttcaa gagatccagc attctgacaa ctcacaaacg catccacacg
      841 ggcgaaaagc cttataaatg cgaagagtgc ggaaaggtct tcaagtatct gtccagcctg
      901 agcacacata agattatcca caccggcgag aagccgtaca agtgcgagga atgcggtaaa
      961 gctttcaact ggtccagcca tctgaccact cacaagagga ttcataccgg agagaagccc
     1021 tataaatgtg aggagtgcgg caagggattc aagtacagca gcaccctgac caaacacaag
     1081 atcatccata ctggagaaaa gccctacaaa tgcgaggagt gtggagaggc cttcaaatac
     1141 tcctgcagcc tgaccgccca taaaatcata cacaccggaa agaaacccta taagtgtgag
     1201 gaatgtggaa aggtgtttaa gcacagcagc cccctgagca aacataagag gatccacact
     1261 ggggagaagc cttacaagtg tgaagagtgt ggtaaagcct ttagcaggag cagcatcctg
     1321 accacacaca aaatcattca tacaggagaa aaaccctacg agtgtgaaga ttgcggcaag
     1381 gcctttaaca gaagcagcaa cctgacaaag cacaagaaaa ttcacacagg agagaaacca
     1441 tacaaatgtg aagagtgcgg gaaagccttc aagtgcagct ctatcctgac tacacataaa
     1501 aggattcaca ccgccgacaa gccatataag tgcgaagaat gcgggaagga ctttaagtac
     1561 tccagcacac tgaccagaca caaaaaaatc cacacaggcg gcaagcccca caagtgtaac
     1621 aaatgcggca aagcattcat cagcagctcc aacctgagca ggcacgagat cattcacatg
     1681 ggcggcaacc cctacaagtg tgaaaacgtg gccaagccct atacccagct ttcttgtaca
     1741 aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca
     1801 gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca
     1861 tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg
     1921 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     1981 caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca
     2041 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     2101 caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     2161 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     2221 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     2281 ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     2341 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     2401 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     2461 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     2521 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
     2581 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     2641 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     2701 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     2761 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt
     2821 tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat
     2881 cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     2941 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3001 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3061 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3121 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3181 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3241 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3301 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3361 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3421 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3481 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3541 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     3601 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     3661 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     3721 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     3781 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta
     3841 gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt
     3901 ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac
     3961 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca
     4021 acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc
     4081 agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgttgtaaaa
     4141 cgacggccag tcttaagctc gggccccaaa taatgatttt attttgactg atagtgacct
     4201 gttcgttgca acaaattgat gagcaatgct tttttataat gccaactttg tacaaaaaag
     4261 caggctta
//
