LOCUS       sequence-203109-        3003 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recipient vector for Golden Gate TALE cloning system. Used to 
            assemble 8 TALE repeats..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3003)
  AUTHORS   Lowder LG, Zhou J, Zhang Y, Malzahn A, Zhong Z, Hsieh TF, Voytas DF,
            Zhang Y, Qi Y
  TITLE     Robust transcriptional activation in plants using multiplexed 
            CRISPR-Act2.0 and mTALE-Act systems.
  JOURNAL   Mol Plant. 2017 Nov 29. pii: S1674-2052(17)30344-1. doi: 
            10.1016/j.molp.2017.11.010.
  PUBMED    29197638
REFERENCE   2  (bases 1 to 3003)
  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: "Mol Plant. 
            2017 Nov 29. pii: S1674-2052(17)30344-1. doi: 
            10.1016/j.molp.2017.11.010."
COMMENT     Sequence Label: pFUS_A8
FEATURES             Location/Qualifiers
     source          1..3003
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(57..434)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLDSH
                     HLVRGQVQGETLKTGRRFTTS"
     primer_bind     67..83
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     299..315
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     325..344
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        325..343
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(391..411)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(391..409)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(430..446)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(430..446)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(443..465)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    454..470
                     /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(478..484,485..502,503..508))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    523..544
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             999..1790
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1883..2471
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2372..2391
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2625..2642
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2801..2828)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(join(2920..3003,1..3))
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac
       61 gacgttgtaa aacgacggcc agtcttaagc gtctccccct gaacctgacc ccggaccaag
      121 tggtggctat cgagaccggc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
      181 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
      241 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
      301 aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt gggtaccggg
      361 ccccccctcg aggtcctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc
      421 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
      481 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
      541 attaattgcg ttgcgctcac tgcccgcttt ccaccggtgg tctcttcgaa acggtgcagc
      601 ggctgttgcc ggtgctgtgc caggaccatg gccgagacgt ctagaccagc caggacagaa
      661 atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg aaacggatga
      721 aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta
      781 tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg
      841 cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca
      901 agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta
      961 cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg gtgatcgccg
     1021 aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt
     1081 tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata
     1141 ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca
     1201 acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag
     1261 tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc
     1321 aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg
     1381 acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc
     1441 cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg
     1501 aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc
     1561 ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg
     1621 ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta
     1681 gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag
     1741 aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc
     1801 acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg
     1861 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1921 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1981 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     2041 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2101 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2161 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2221 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2281 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2341 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2401 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     2461 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     2521 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     2581 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     2641 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     2701 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     2761 atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat
     2821 ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc
     2881 gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc
     2941 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt
     3001 tat
//
