LOCUS       sequence-191593-        6038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor Vector containing ZNF516 transcription factor, part of the 
            Human TFome Collection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6038)
  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 6038)
  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-ZNF516
FEATURES             Location/Qualifiers
     source          1..6038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2373..2382
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3493..3592)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3609..3628)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3610..3628)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3633..3649)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3633..3649)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3751..3770)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3762..4571
                     /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(3829..3848)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4718..5306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5207..5226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5460..5477
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5636..5663)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5755..5841)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5890..5912
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5904..5921
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5905..5921
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5937..6036
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 atggacagaa acagagaagc agaaatggaa ttgagaagag gaccttcacc tacaagggct
       61 ggaagaggcc atgaagtaga tggtgataaa gctacgtgcc atacctgctg tatctgtgga
      121 aagagtttcc ctttccaatc aagtttaagc caacatatga ggaaacatac tggagaaaag
      181 ccatataaat gcccgtactg tgaccaccgg gcaagccaaa aaggaaacct aaaaatccat
      241 ataagaagcc acaggactgg cactttaatc caaggacatg agccagaggc tggtgaagcc
      301 cctttaggtg aaatgagggc cagtgaaggc ctagatgcgt gtgcgagccc cacaaagagt
      361 gcatcggcat gtaacagact tttaaatgga gcttcccagg ctgatggagc gagagtttta
      421 aacggtgcca gccaggcaga cagcgggaga gtccttttga gatcttcaaa aaaaggtgca
      481 gaagggagtg catgtgcccc aggtgaagca aaagcagctg tccaatgttc tttctgtaag
      541 agccaatttg aaagaaaaaa agaccttgaa ctccatgtcc accaggccca taaacctttt
      601 aagtgcagac tatgttcata tgcaactttg agagaagaaa gccttttatc ccatatagag
      661 agagatcata tcacggccca aggccctgga agtggagagg cctgtgtaga aaatggaaag
      721 ccagaattat cgcctggaga attcccctgt gaagtctgtg gccaagcttt cagccagacg
      781 tggttcctaa aggcacatat gaagaaacac agaggcagtt ttgaccatgg atgccatata
      841 tgtgggagga gatttaaaga gccatggttc ctgaagaacc atatgaaagc ccatggacca
      901 aagactggtt caaagaacag gccaaaatct gaattagacc caattgctac aataaataat
      961 gttgtccagg aagaagtaat cgtggctggc ctcagcctat atgaagtgtg tgcaaaatgt
     1021 ggaaaccttt ttacaaacct tgattcttta aatgcacata atgcaatcca cagaagagtt
     1081 gaagccagca ggaccagggc gcctgctgaa gaaggagcag aaggccccag tgacacaaaa
     1141 caatttttcc tccagtgcct aaacctcagg ccaagtgcag ctggtgattc gtgccctggt
     1201 acccaagctg gaaggagagt tgcagaacta gaccctgtaa attcatacca ggcatggcaa
     1261 ttggctacca gagggaaagt agcagagcct gcagaatacc taaaatatgg ggcgtgggat
     1321 gaagctttag ccggagatgt tgcttttgat aaagacagaa gagaatatgt ccttgtaagc
     1381 caagaaaaga ggaagagaga acaagatgcc cctgcggcac aaggaccccc cagaaaaaga
     1441 gcatctggcc ctggtgaccc tgccccagct ggacaccttg acccaagatc agcagcgagg
     1501 ccaaacagaa gggctgcggc aaccactggc caaggaaaat ccagtgaatg ttttgagtgt
     1561 ggaaaaatat tcaggacata ccaccaaatg gtcctacaca gcagagtcca caggagagcg
     1621 aggagagaaa gagattctga tggtgacagg gcagcgagag caagatgtgg aagcctgagt
     1681 gaaggagatt cagcgagcca accatcgagc cctggatcgg cgtgtgctgc agctgacagc
     1741 cctggttctg gcctagctga tgaggcagca gaagacagtg gagaagaagg tgcacctgag
     1801 cctgcgccag gtggccagcc caggagatgc tgtttcagtg aagaagtcac atctactgaa
     1861 ttaagcagtg gtgaccagag ccataaaatg ggagataatg ccagtgagag agacactggt
     1921 gaatcaaagg cagggatagc tgccagtgta tcgatccttg aaaattcatc gagagaaaca
     1981 tccaggagac aagaacaaca cagattttca atggacctga aaatgcctgc tttccacccg
     2041 aaacaagaag tccctgtgcc tggtgatggt gttgaattcc catcgagtac aggggcagaa
     2101 ggacagacag gccacccagc agaaaaatta tctgacctcc ataataaaga acacagcgga
     2161 ggaggaaaga gggctttggc cccagaccta atgccccttg acctcagtgc gaggagcacc
     2221 agagatgacc cttcgaataa agaaactgca tcttccctcc aggcagccct tgttgtccac
     2281 ccatgcccat actgttccca taagacatat tacccagaag ttttgtggat gcataaaaga
     2341 atctggcaca gagtttcatg taattctgtt gcgccgccat ggatccagcc aaatggatat
     2401 aaatcaatca gatcaaacct tgtatttttg agcagatcag ggagaactgg cccgccgccg
     2461 gccctcggtg gaaaagaatg ccaacccctc ctcctggcga gatttacgag aacccaagta
     2521 ccaggtggaa tgcctggatc aaaaagtgga tcatcgcccc ttggtgttgt gacaaaggcg
     2581 gcttcaatgc caaaaaataa agagagccac agtggtggcc cgtgtgccct gtgggcacct
     2641 gggcctgatg gatacagaca gacaaaaccg tgccatggcc aagaacccca tggagcagca
     2701 acccaaggac ctttggcaaa gcccagacaa gaggcttcat caaagcctgt cccggcgcct
     2761 ggcggtggtg gtttttcaag aagtgcaacg ccgacgccga cagtaattgc gagggcaggg
     2821 gcccagccga gtgcaaattc gaagcctgtt gaaaaatttg gtgtcccccc ggctggggct
     2881 ggttttgccc ctacaaataa acacagtgcg cctgacagtt taaaagcaaa attttcggcc
     2941 caaccccaag ggccaccgcc ggcaaaagga gaaggtggag cgccaccttt accacccaga
     3001 gaaccccctt caaaggctgc ccaagaattg aggactttag ccacatgtgc tgctggatcg
     3061 agaggtgatg cggccctcca agcccagcct ggtgttgctg gtgcaccccc tgttttacat
     3121 tcaataaaac aagagccggt ggcggaaggc catgagaaga gattagatat tttgaacatt
     3181 tttaagacgt atatcccgaa agattttgcc accctatacc aaggatgggg tgtaagtgga
     3241 cctggccttg aacacagagg gaccctgagg acccaggcca gaccaggaga atttgtatgt
     3301 atagaatgtg ggaaaagttt ccaccaacct ggccatttga gggcccatat gagagcacac
     3361 agtgttgttt ttgaatcaga tggaccgaga gggagtgaag tccacactac gagtgcagat
     3421 gcaccaaaac aaggaaggga ccattcaaac actggaactg tccagactgt acccctaaga
     3481 aaaggtacat atacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat
     3541 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag
     3601 ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc
     3661 ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa
     3721 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg
     3781 aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     3841 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg
     3901 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     3961 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     4021 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg
     4081 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     4141 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     4201 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     4261 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct
     4321 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     4381 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     4441 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     4501 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     4561 agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt
     4621 gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact
     4681 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4741 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4801 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4861 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4921 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4981 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5041 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5101 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5161 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5221 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5281 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5341 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5401 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     5461 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     5521 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     5581 agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag
     5641 gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc
     5701 tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt
     5761 cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga
     5821 ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcgtta acgctagcat
     5881 ggatgttttc ccagtcacga cgttgtaaaa cgacggccag tcttaagctc gggccccaaa
     5941 taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat gagcaatgct
     6001 tttttataat gccaactttg tacaaaaaag caggctta
//
