LOCUS       sequence-75703-e        7363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TALEN expression vector for yeast, N152,C18 architecture.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7363)
  AUTHORS   Zhang Y, Zhang F, Li X, Baller JA, Qi Y, Starker CG, Bogdanove AJ, 
            Voytas DF
  TITLE     TALENs enable efficient plant genome engineering.
  JOURNAL   Plant Physiol. 2012 Nov 2.
  PUBMED    23124327
REFERENCE   2  (bases 1 to 7363)
  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: "Plant 
            Physiol. 2012 Nov 2."
COMMENT     Sequence Label: pZHY566
FEATURES             Location/Qualifiers
     source          1..7363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        286..690
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             703..1797
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     misc_feature    2226..2729
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        2771..2875
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3943..4531
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4432..4451
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4685..4702
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      4803..4897
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        4904..4922
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        4999..5397
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             5469..5489
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5490..5516
                     /codon_start=1
                     /product="epitope tag from the baculovirus Autographa 
                     californica GP64 envelope fusion protein"
                     /label=AcV5 tag
                     /translation="SWKDASGWS"
     CDS             complement(5823..6230)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG
                     TRQCIARCMHCLHCGHAATFCNLHKLACVQL"
     primer_bind     6080..6102
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6094..6111
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6095..6111
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6121..6139
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6187..6205)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6226..6242)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6226..6242)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6239..6261)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6250..6266
                     /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)."
     promoter        complement(join(6274..6280,6281..6298,6299..6304))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6319..6340
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             6391..6978
                     /codon_start=1
                     /product="nonspecific DNA cleavage domain of the FokI 
                     endonuclease (Li et al., 1992)"
                     /label=FokI cleavage domain
                     /note="must dimerize to be catalytically active"
                     /translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
                     MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
                     YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     terminator      7001..7253
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(7295..7313)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      complement(7315..7354)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
ORIGIN
        1 ttctccttac gcatctgtgc ggtatttcac accgcatatc gacggtcgag gagaacttct
       61 agtatatcca catacctaat attattgcct tattaaaaat ggaatcccaa caattacatc
      121 aaaatccaca ttctcttcaa aatcaattgt cctgtacttc cttgttcatg tgtgttcaaa
      181 aacgttatat ttataggata attatactct atttctcaac aagtaattgg ttgtttggcc
      241 gagcggtcta aggcgcctga ttcaagaaat atcttgaccg cagttaactg tgggaatact
      301 caggtatcgt aagatgcaag agttcgaatc tcttagcaac cattattttt ttcctcaaca
      361 taacgagaac acacaggggc gctatcgcac agaatcaaat tcgatgactg gaaatttttt
      421 gttaatttca gaggtcgcct gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa
      481 ggaaaggtga gaggccggaa ccggcttttc atatagaata gagaagcgtt catgactaaa
      541 tgcttgcatc acaatacttg aagttgacaa tattatttaa ggacctattg ttttttccaa
      601 taggtggtta gcaatcgtct tactttctaa cttttcttac cttttacatt tcagcaatat
      661 atatatatat ttcaaggata taccattcta atgtctgccc ctatgtctgc ccctaagaag
      721 atcgtcgttt tgccaggtga ccacgttggt caagaaatca cagccgaagc cattaaggtt
      781 cttaaagcta tttctgatgt tcgttccaat gtcaagttcg atttcgaaaa tcatttaatt
      841 ggtggtgctg ctatcgatgc tacaggtgtc ccacttccag atgaggcgct ggaagcctcc
      901 aagaaggttg atgccgtttt gttaggtgct gtgggtggtc ctaaatgggg taccggtagt
      961 gttagacctg aacaaggttt actaaaaatc cgtaaagaac ttcaattgta cgccaactta
     1021 agaccatgta actttgcatc cgactctctt ttagacttat ctccaatcaa gccacaattt
     1081 gctaaaggta ctgacttcgt tgttgtcaga gaattagtgg gaggtattta ctttggtaag
     1141 agaaaggaag acgatggtga tggtgtcgct tgggatagtg aacaatacac cgttccagaa
     1201 gtgcaaagaa tcacaagaat ggccgctttc atggccctac aacatgagcc accattgcct
     1261 atttggtcct tggataaagc taatgttttg gcctcttcaa gattatggag aaaaactgtg
     1321 gaggaaacca tcaagaacga attccctaca ttgaaggttc aacatcaatt gattgattct
     1381 gccgccatga tcctagttaa gaacccaacc cacctaaatg gtattataat caccagcaac
     1441 atgtttggtg atatcatctc cgatgaagcc tccgttatcc caggttcctt gggtttgttg
     1501 ccatctgcgt ccttggcctc tttgccagac aagaacaccg catttggttt gtacgaacca
     1561 tgccacggtt ctgctccaga tttgccaaag aataaggttg accctatcgc cactatcttg
     1621 tctgctgcaa tgatgttgaa attgtcattg aacttgcctg aagaaggtaa ggccattgaa
     1681 gatgcagtta aaaaggtttt ggatgcaggt atcagaactg gtgatttagg tggttccaac
     1741 agtaccaccg aagtcggtga tgctgtcgcc gaagaagtta agaaaatcct tgcttaaaaa
     1801 gattctcttt ttttatgata tttgtacata aactttataa atgaaattca taatagaaac
     1861 gacacgaaat tacaaaatgg aatatgttca tagggtagac gaaactatat acgcaatcta
     1921 catacattta tcaagaagga gaaaaaggag gatagtaaag gaatacaggt aagcaaattg
     1981 atactaatgg ctcaacgtga taaggaaaaa gaattgcact ttaacattaa tattgacaag
     2041 gaggagggca ccacacaaaa agttaggtgt aacagaaaat catgaaacta cgattcctaa
     2101 tttgatattg gaggattttc tctaaaaaaa aaaaaataca acaaataaaa aacactcaat
     2161 gacctgacca tttgatgggc tcaccaagct cttaaaacgg gaatttatgg atcttaatta
     2221 agacggatcg cttgcctgta acttacacgc gcctcgtatc ttttaatgat ggaataattt
     2281 gggaatttac tctgtgttta tttattttta tgttttgtat ttggatttta gaaagtaaat
     2341 aaagaaggta gaagagttac ggaatgaaga aaaaaaaata aacaaaggtt taaaaaattt
     2401 caacaaaaag cgtactttac atatatattt attagacaag aaaagcagat taaatagata
     2461 tacattcgat taacgataag taaaatgtaa aatcacagga ttttcgtgtg tggtcttcta
     2521 cacagacaag atgaaacaat tcggcattaa tacctgagag caggaagagc aagataaaag
     2581 gtagtatttg ttggcgatcc ccctagagtc ttttacatct tcggaaaaca aaaactattt
     2641 tttctttaat ttcttttttt actttctatt tttaatttat atatttatat taaaaaattt
     2701 aaattataat tatttttata gcacgtgatg aaaagcctta gggtcaggtg gcacttttcg
     2761 gggaaatgtg cgatgaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2821 gctaatgaga caataaccct gataaatgct tcaattatat tgaaaaagga agagtatgag
     2881 tatccaacat ttccgtgtcg cccttattcc cttttttgcg gccttctgcc ttcctgtttt
     2941 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcccgagt
     3001 gggttacatt gaactggatc tcaacagcgg caaaatctta gagagttttc gccccgaaga
     3061 acgcttccca atgatgagca ctttcaaagt tctgctatgt ggcgcggtgt tatcccgtat
     3121 tgatgctggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3181 gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgtag
     3241 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg
     3301 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3361 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3421 agcaatggca acaactctcc gcaaactatt aactggcgaa ctacttactc ttgcttcacg
     3481 ccagcaactc attgactgga tggaggcgga caaagttgca ggaccacttc tgcgctcggc
     3541 acttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3601 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3661 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3721 gattaagcat tggtaactgt cagacgaagg gaaataaata gtagcccgcc tgatgtgcgg
     3781 gctttttttt ggtggtggtt accagtgtcg cttagctacc gtaggtcctc ttaggcggtc
     3841 cgattagcga catagtcatc aatgccttga cggcaaaatc ccttaacgtg agttttcgtt
     3901 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3961 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4021 ggatcaagag ctactaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4081 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4141 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4201 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4261 aacggggggt tcgtgctcac agcccagctt ggagcgaacg acctacaccg aactgagata
     4321 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4381 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4441 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4501 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4561 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     4621 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     4681 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     4741 cgcgcgttgg ccgattcatt gatgcaggtc aacgacatca cctggttgcg agccttctga
     4801 aggactcctg ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac
     4861 gctcggttgc cgccgggcgt tttttattgg tgagaatcag gcgtaatacg actcactata
     4921 gggcgaatgg aactggacgc tcgcctgtgg tcgcagggtg gctcgtacga acttaccgcg
     4981 gagacatatc gataatcaca tagcttcaaa atgtttctac tcctttttta ctcttccaga
     5041 ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca gcatactaaa
     5101 tttcccctct ttcttcctct agggtgtcgt taattacccg tactaaaggt ttggaaaaga
     5161 aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac
     5221 gtttcttttt cttgaaaatt ttttttttga tttttttctc tttcgatgac ctcccattga
     5281 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
     5341 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
     5401 agatctaact cgagaaagat attgtatata tcgtaacaat aggaggttca acaatggctt
     5461 cctcccctcc aaagaaaaag agaaaggtta gttggaagga cgcaagtggt tggtctagaa
     5521 tggtggatct acgcacgctc ggctacagtc agcagcagca agagaagatc aaaccgaagg
     5581 tgcgttcgac agtggcgcag caccacgagg cactggtggg ccatgggttt acacacgcgc
     5641 acatcgttgc gctcagccaa cacccggcag cgttagggac cgtcgctgtc acgtatcagc
     5701 acataatcac ggcgttgcca gaggcgacac acgaagacat cgttggcgtc ggcaaacagt
     5761 ggtccggcgc acgcgccctg gaggccttgc tcacggatgc gggggagttg agaggtccgc
     5821 cgttacagtt ggacacaggc caacttgtga agattgcaaa acgtggcggc gtgaccgcaa
     5881 tggaggcagt gcatgcatcg cgcaatgcac tgacgggtgc ccccctggag acgggcgccg
     5941 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     6001 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     6061 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg
     6121 taatacgact cactataggg cgaattgggt accgggcccc ccctcgaggt cctccagctt
     6181 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     6241 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6301 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6361 cgctttccac cggtcgtctc caacggatcc cagctagtga aatctgaatt ggaagagaag
     6421 aaatctgaac ttagacataa attgaaatat gtgccacatg aatatattga attgattgaa
     6481 atcgcaagaa attcaactca ggatagaatc cttgaaatga aggtgatgga gttctttatg
     6541 aaggtttatg gttatcgtgg taaacatttg ggtggatcaa ggaaaccaga cggagcaatt
     6601 tatactgtcg gatctcctat tgattacggt gtgatcgttg atactaaggc atattcagga
     6661 ggttataatc ttccaattgg tcaagcagat gaaatgcaaa gatatgtcga agagaatcaa
     6721 acaagaaaca agcatatcaa ccctaatgaa tggtggaaag tctatccatc ttcagtaaca
     6781 gaatttaagt tcttgtttgt gagtggtcat ttcaaaggaa actacaaagc tcagcttaca
     6841 agattgaatc atatcactaa ttgtaatgga gctgttctta gtgtagaaga gcttttgatt
     6901 ggtggagaaa tgattaaagc tggtacattg acacttgagg aagtgagaag gaaatttaat
     6961 aacggtgaga taaactttta ataggagctc gaatttcccc gatcgttcaa acatttggca
     7021 ataaagtttc ttaagattga atcctgttgc cggtcttgcg atgattatca tataatttct
     7081 gttgaattac gttaagcatg taataattaa catgtaatgc atgacgttat ttatgagatg
     7141 ggtttttatg attagagtcc cgcaattata catttaatac gcgatagaaa acaaaatata
     7201 gcgcgcaaac taggataaat tatcgcgcgc ggtgtcatct atgttactag atcgtgggcc
     7261 cacattcctg tagaactcgg tgagactttt gttcccttta gtgagggtta attgtccaaa
     7321 aaaaaaggct ccaaaaggag cctttaattg tttcggttta tgc
//
