LOCUS       sequence-75702-e        6426 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 6426)
  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 6426)
  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: pZHY565
FEATURES             Location/Qualifiers
     source          1..6426
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      35..129
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        136..154
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        231..629
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             701..721
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             722..748
                     /codon_start=1
                     /product="epitope tag from the baculovirus Autographa 
                     californica GP64 envelope fusion protein"
                     /label=AcV5 tag
                     /translation="SWKDASGWS"
     CDS             complement(1055..1462)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG
                     TRQCIARCMHCLHCGHAATFCNLHKLACVQL"
     primer_bind     1312..1334
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1326..1343
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1327..1343
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1353..1371
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1419..1437)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1458..1474)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1458..1474)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1471..1493)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1482..1498
                     /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(1506..1512,1513..1530,1531..1536))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1551..1572
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1623..2210
                     /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      2233..2485
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(2527..2545)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      complement(2547..2586)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(2886..3545)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(3546..3733)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     misc_feature    3884..4387
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4429..4533
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5601..6189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6090..6109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6343..6360
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcaacgacat cacctggttg cgagccttct gaaggactcc tgttgataga tccagtaatg
       61 acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt
      121 ggtgagaatc aggcgtaata cgactcacta tagggcgaat ggaactggac gctcgcctgt
      181 ggtcgcaggg tggctcgtac gaacttaccg cggagacata tcgataatca catagcttca
      241 aaatgtttct actccttttt tactcttcca gattttctcg gactccgcgc atcgccgtac
      301 cacttcaaaa cacccaagca cagcatacta aatttcccct ctttcttcct ctagggtgtc
      361 gttaattacc cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt ttctttttct
      421 tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt ttcttgaaaa tttttttttt
      481 gatttttttc tctttcgatg acctcccatt gatatttaag ttaataaacg gtcttcaatt
      541 tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt cttgctcatt
      601 agaaagaaag catagcaatc taatctaagt ttagatctaa ctcgagaaag atattgtata
      661 tatcgtaaca ataggaggtt caacaatggc ttcctcccct ccaaagaaaa agagaaaggt
      721 tagttggaag gacgcaagtg gttggtctag aatggtggat ctacgcacgc tcggctacag
      781 tcagcagcag caagagaaga tcaaaccgaa ggtgcgttcg acagtggcgc agcaccacga
      841 ggcactggtg ggccatgggt ttacacacgc gcacatcgtt gcgctcagcc aacacccggc
      901 agcgttaggg accgtcgctg tcacgtatca gcacataatc acggcgttgc cagaggcgac
      961 acacgaagac atcgttggcg tcggcaaaca gtggtccggc gcacgcgccc tggaggcctt
     1021 gctcacggat gcgggggagt tgagaggtcc gccgttacag ttggacacag gccaacttgt
     1081 gaagattgca aaacgtggcg gcgtgaccgc aatggaggca gtgcatgcat cgcgcaatgc
     1141 actgacgggt gcccccctgg agacgggcgc cgctacaggg cgcgtcccat tcgccattca
     1201 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     1261 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     1321 gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg
     1381 gtaccgggcc ccccctcgag gtcctccagc ttttgttccc tttagtgagg gttaattgcg
     1441 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     1501 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     1561 taactcacat taattgcgtt gcgctcactg cccgctttcc accggtcgtc tccaacggat
     1621 cccagctagt gaaatctgaa ttggaagaga agaaatctga acttagacat aaattgaaat
     1681 atgtgccaca tgaatatatt gaattgattg aaatcgcaag aaattcaact caggatagaa
     1741 tccttgaaat gaaggtgatg gagttcttta tgaaggttta tggttatcgt ggtaaacatt
     1801 tgggtggatc aaggaaacca gacggagcaa tttatactgt cggatctcct attgattacg
     1861 gtgtgatcgt tgatactaag gcatattcag gaggttataa tcttccaatt ggtcaagcag
     1921 atgaaatgca aagatatgtc gaagagaatc aaacaagaaa caagcatatc aaccctaatg
     1981 aatggtggaa agtctatcca tcttcagtaa cagaatttaa gttcttgttt gtgagtggtc
     2041 atttcaaagg aaactacaaa gctcagctta caagattgaa tcatatcact aattgtaatg
     2101 gagctgttct tagtgtagaa gagcttttga ttggtggaga aatgattaaa gctggtacat
     2161 tgacacttga ggaagtgaga aggaaattta ataacggtga gataaacttt taataggagc
     2221 tcgaatttcc ccgatcgttc aaacatttgg caataaagtt tcttaagatt gaatcctgtt
     2281 gccggtcttg cgatgattat catataattt ctgttgaatt acgttaagca tgtaataatt
     2341 aacatgtaat gcatgacgtt atttatgaga tgggttttta tgattagagt cccgcaatta
     2401 tacatttaat acgcgataga aaacaaaata tagcgcgcaa actaggataa attatcgcgc
     2461 gcggtgtcat ctatgttact agatcgtggg cccacattcc tgtagaactc ggtgagactt
     2521 ttgttccctt tagtgagggt taattgtcca aaaaaaaagg ctccaaaagg agcctttaat
     2581 tgtttcggtt tatgcttagg acaccattga ttaacgctta caatttcctg atgcggtatt
     2641 ttctccttac gcatctgtgc ggtatttcac accgcataga tccgtcgagt tcaagagaaa
     2701 aaaaaagaaa aagcaaaaag aaaaaaggaa agcgcgcctc gttcagaatg acacgtatag
     2761 aatgatgcat taccttgtca tcttcagtat catactgttc gtataaatac ttactgacat
     2821 tcatagctat acatatatac acatgtatat atatcgtatg ctggagcttt aaataatcgg
     2881 tgtcactaca taagaacacc tttggtggag ggaacatcgt tggttccatt gggcgaggtg
     2941 gcttctctta tggcaaccgc aagagccttg aacgcactct cactacggtg atggtcattc
     3001 ttgcctcgca gacaatcaac gtggagggta attctgcttg cctctgcaaa actttcaaga
     3061 aaatgcggga tcatctcgca agagaggtct cctactttct ccctttgcaa accaagttcg
     3121 acaactgcgt atggcctgtt agaaaggtct accaccgctc tggaaagtgc ctcatccaaa
     3181 ggcgcaaatc ctgatccaaa cctttttact ccacgcgcca gtagagcctc tttaaaagcc
     3241 tgaccgagag caatcccgca gtcttcagtg gtgtgatggt cgtctatgtg taagtcacca
     3301 atgcactcaa cgattagcga ccagccggaa tgcttggcca gagcatgtat catgtggtcc
     3361 agaaacccta tacctgtgtg gacgttaatc acttgcgatt gtgtggcctg ttctgctact
     3421 gcttctgcct ctttttctgg gaagatcgag tgctctatcg ctaggggacc accctttaaa
     3481 gagatagcaa tctgaatctt ggtttcattt gtaatacgct ttactagggc tttctgctct
     3541 gtcatctttg ccttcgttta tcttgcctgc tcatttttta gtatattctt cgaagaaatc
     3601 acattacttt atataatgta taattcatta tgtgataatg ccaatcgcta agaaaaaaaa
     3661 agagtcatcc gctaggtgga aaaaaaaaaa tgaaaatcat taccgaggca taaaaaaata
     3721 tagagtgtac tagaggaggc caagagtaat agaaaaagaa aattgcggga aaggactgtg
     3781 ttatgacttc cctgactaat gccgtgttca aacgatacct ggcagtgact cctagcgctc
     3841 accaagctct taaaacggga atttatggat cttaattaag acggatcgct tgcctgtaac
     3901 ttacacgcgc ctcgtatctt ttaatgatgg aataatttgg gaatttactc tgtgtttatt
     3961 tatttttatg ttttgtattt ggattttaga aagtaaataa agaaggtaga agagttacgg
     4021 aatgaagaaa aaaaaataaa caaaggttta aaaaatttca acaaaaagcg tactttacat
     4081 atatatttat tagacaagaa aagcagatta aatagatata cattcgatta acgataagta
     4141 aaatgtaaaa tcacaggatt ttcgtgtgtg gtcttctaca cagacaagat gaaacaattc
     4201 ggcattaata cctgagagca ggaagagcaa gataaaaggt agtatttgtt ggcgatcccc
     4261 ctagagtctt ttacatcttc ggaaaacaaa aactattttt tctttaattt ctttttttac
     4321 tttctatttt taatttatat atttatatta aaaaatttaa attataatta tttttatagc
     4381 acgtgatgaa aagccttagg gtcaggtggc acttttcggg gaaatgtgcg atgaacccct
     4441 atttgtttat ttttctaaat acattcaaat atgtatccgc taatgagaca ataaccctga
     4501 taaatgcttc aattatattg aaaaaggaag agtatgagta tccaacattt ccgtgtcgcc
     4561 cttattccct tttttgcggc cttctgcctt cctgtttttg ctcacccaga aacgctggtg
     4621 aaagtaaaag atgctgaaga tcagttgggt gcccgagtgg gttacattga actggatctc
     4681 aacagcggca aaatcttaga gagttttcgc cccgaagaac gcttcccaat gatgagcact
     4741 ttcaaagttc tgctatgtgg cgcggtgtta tcccgtattg atgctgggca agagcaactc
     4801 ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag
     4861 catcttacgg atggcatgac agtaagagaa ttatgtagtg ctgccataac catgagtgat
     4921 aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt
     4981 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5041 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aactctccgc
     5101 aaactattaa ctggcgaact acttactctt gcttcacgcc agcaactcat tgactggatg
     5161 gaggcggaca aagttgcagg accacttctg cgctcggcac ttccggctgg ctggtttatt
     5221 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5281 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5341 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5401 gacgaaggga aataaatagt agcccgcctg atgtgcgggc ttttttttgg tggtggttac
     5461 cagtgtcgct tagctaccgt aggtcctctt aggcggtccg attagcgaca tagtcatcaa
     5521 tgccttgacg gcaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5581 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5641 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct actaactctt
     5701 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5761 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5821 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5881 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgctcacag
     5941 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6001 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6061 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6121 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6181 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6241 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6301 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6361 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattga
     6421 tgcagg
//
