LOCUS       40924_43139        6567 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Moss transformation vector pTFH22.4 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6567)
  AUTHORS   Sakakibara K, Reisewitz P, Aoyama T, Friedrich T, Ando S, Sato Y, 
            Nishiyama T, Tamada Y, Hiwatashi Y, Kurata T, Ishikawa M, Deguchi H,
            Rensing SA, Werr W, Murata T, Hasebe M, Laux T.
  TITLE     WOX13-like genes are required for the initiation of apical stem 
            cells from differentiated leaf cells in the moss Physcomitrella 
            patens
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6567)
  AUTHORS   Sakakibara K, Fujita T, Hasebe M.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-OCT-2012) Contact:Keiko Sakakibara Graduate School of 
            Science, Hiroshima University, Department of Biological Science; 
            1-3-1 Kagamiyama, Higashi-Hiroshima, Hiroshima 739-8526, Japan
REFERENCE   3  (bases 1 to 6567)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6567)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-OCT-2012) Contact:Keiko Sakakibara Graduate School of Science, 
            Hiroshima University, Department of Biological Science; 1-3-1 
            Kagamiyama, Higashi-Hiroshima, Hiroshima 739-8526, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      28..616
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    904..925
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        940..970
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    978..994
                     /label=lac operator
                     /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)."
     primer_bind     1002..1018
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1055..1893
                     /label=Act1 promoter
                     /note="rice actin promoter"
     CDS             1898..1915
                     /label=6xHis
                     /note="6xHis affinity tag"
     misc_feature    2450..2976
                     /label=TrbcS
                     /note="TrbcS"
     misc_feature    2461..2496
                     /note="SfiI Multi cloning site (SfiI, AscI, SmaI, ApaI,
                     SfiI)"
     promoter        3055..3402
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             3423..4214
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4287..4462
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        5143..5488
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             5536..6252
                     /label=EGFP
                     /note="enhanced GFP"
     terminator      6263..6515
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
ORIGIN
        1 actagtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
       61 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
      121 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
      181 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
      241 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
      301 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
      361 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
      421 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
      481 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
      541 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
      601 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
      661 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
      721 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
      781 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
      841 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
      901 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
      961 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     1021 tgattacgaa ttcgagctcg gtacccctca agcttcgagg tcattcatat gcttgagaag
     1081 agagtcggga tagtccaaaa taaaacaaag gtaagattac ctggtcaaaa gtgaaaacat
     1141 cagttaaaag gtggtataaa gtaaaatatc ggtaataaaa ggtggcccaa agtgaaattt
     1201 actcttttct actattataa aaattgagga tgtttttgtc ggtactttga tacgtcattt
     1261 ttgtatgaat tggtttttaa gtttattcgc ttttggaaat gcatatctgt atttgagtcg
     1321 ggttttaagt tcgtttgctt ttgtaaatac agagggattt gtataagaaa tatctttaaa
     1381 aaaacccata tgctaatttg acataatttt tgagaaaaat atatattcag gcgaattctc
     1441 acaatgaaca ataataagat taaaatagct ttcccccgtt gcagcgcatg ggtatttttt
     1501 ctagtaaaaa taaaagataa acttagactc aaaacattta caaaaacaac ccctaaagtt
     1561 cctaaagccc aaagtgctat ccacgatcca tagcaagccc agcccaaccc aacccaaccc
     1621 aacccacccc agtccagcca actggacaat agtctccaca cccccccact atcaccgtga
     1681 gttgtccgca cgcaccgcac gtctcgcagc caaaaaaaaa aaagaaagaa aaaaaagaaa
     1741 aagaaaaaac agcaggtggg tccgggtcgt gggggccgga aacgcgagga ggatcgcgag
     1801 ccagcgacag gccggccctc cctccgcttc caaagaaacg ccccccatcg ccactatata
     1861 catccccccc ctctcctccc atccccccaa cccctaccac caccaccacc accacctcca
     1921 cctcctcccc cctcgctgcc ggacgacgag ctcctccccc ctccccctcc gccgccgccg
     1981 cgccggtaac caccccgccc ctctcctctt tctttctccg tttttttttt ccgtctcggt
     2041 ctcgatcttt ggccttggta gtttgggtgg gcgagaggcg gcttcgtgcg cgcccagatc
     2101 ggtgcgcggg aggggcggga tctcgcggct gggtctcgcc ggcgtggatc cggcccggat
     2161 ctcgcgggga atggggctct cggatgtaga tctgcgatcc gccgttgttg ggggagatga
     2221 tggggggttt aaaatttccg ccatgctaaa caagatcagg aagaggggaa aagggcacta
     2281 tggtttatat ttttatatat ttctgctgct tcgtcaggct tagatgtgct agatctttct
     2341 ttcttctttt tgtgggtaga atttgaatcc ctcagcattg ttcatcggta gtttttcttt
     2401 tcatgatttg tgacaaatgc agcctcgtgc ggagcttttt tgtaggtaga agatatcggc
     2461 cattcaggcc ggcgcgccac ccgggagggc ccggccgatt tggccatcag ctttcgtccg
     2521 tatcatcggt ttcgacaacg ttcgtcaagt tcaatgcatc agtttcattg cccacacacc
     2581 agaatcctac taagtttgag tattatggca ttggaaaagc tgttttcttc tatcatttgt
     2641 tctgcttgta atttactgtg ttctttcagt ttttgttttc ggacatcaaa atgcaaatgg
     2701 atggataaga gttaataaat gatatggtcc ttttgttcat tctcaaatta ttattatctg
     2761 ttgtttttac tttaatgggt tgaatttaag taagaaagga actaacagtg tgatattaag
     2821 gtgcaatgtt agacatataa aacagtcttt cacctctctt tggttatgtc ttgaattggt
     2881 ttgtttcttc acttatctgt gtaatcaagt ttactatgag tctatgatca agtaattatg
     2941 caatcaagtt aagtacagta taggcttttt gtgtcgacat ttaaatatcg aattcctgca
     3001 gcccccctac tccaaaaatg tcaaagatac agtctcagaa gaccaaaggg ctattgacac
     3061 ttttcaacaa agggtatata atcgggaaga ctcctcggat tccattgccc agctatctgt
     3121 cacttcatcg aaaggacagt agaaaaggaa ggtggctcct acaaatgcca tcattgcgat
     3181 aaaggaaagg ctatcattca agatgcctct gccgacagtg gtcccaaaga tggaccccca
     3241 cccacgagga gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat
     3301 tgatgtgaca tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac
     3361 ccttcctcta tataaggaag ttcatttcat ttggagagga cagcccaagc aaggaggtga
     3421 gcatgattga acaagacgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3481 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3541 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3601 tccaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     3661 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     3721 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     3781 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     3841 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     3901 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cggatgcccg
     3961 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4021 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4081 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4141 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4201 ttgacgagtt cttctgagcg ggactctggg gttcggacgg tacgctgaaa tcaccagtct
     4261 ctctctacaa atctatctct ctctattttc tcataaataa tgtgtgagta gtttcccgat
     4321 aagggaaatt agggttctta tagggtttcg ctcatgtgtt gagcatataa gaaaccctta
     4381 gtatgtattt gtatttgtaa aatacttcta tcaataaaat ttctaattcc taaaaccaaa
     4441 atccagtact aaaatccaga tctcctaaag tccctataga tctttgtcgt gaatataaac
     4501 cagacacgag acgactaaac ctggagccca gacgccgttc gaagctagaa gtaccgctta
     4561 ggcaggaggc cgttagggaa aagatgctaa ggcagggttg gttacgttat cggggatcct
     4621 ctagagtcga cctgcaggca tgcctgcagg tccccagatt agccttttca atttcagaaa
     4681 gaatgctaac ccacagatgg ttagagaggc ttacgcagca ggtctcatca agacgatcta
     4741 cccgagcaat aatctccagg aaatcaaata ccttcccaag aaggttaaag atgcagtcaa
     4801 aagattcagg actaactgca tcaagaacac agagaaagat atatttctca agatcagaag
     4861 tactattcca gtatggacga ttcaaggctt gcttcacaaa ccaaggcaag taatagagat
     4921 tggagtctct aaaaaggtag ttcccactga atcaaaggcc atggagtcaa agattcaaat
     4981 agaggaccta acagaactcg ccgtaaagac tggcgaacag ttcatacaga gtctcttacg
     5041 actcaatgac aagaagaaaa tcttcgtcaa catggtggag cacgacacac ttgtctactc
     5101 caaaaatatc aaagatacag tctcagaaga ccaaagggca attgagactt ttcaacaaag
     5161 ggtaatatcc ggaaacctcc tcggattcca ttgcccagct atctgtcact ttattgtgaa
     5221 gatagtggaa aaggaaggtg gctcctacaa atgccatcat tgcgataaag gaaaggccat
     5281 cgttgaagat gcctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat
     5341 cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgatatctc
     5401 cactgacgta agggatgacg cacaatccca ctatccttcg caagaccctt cctctatata
     5461 aggaagttca tttcatttgg agagaacacg ggggactcta gaggatcccc gtggcaggtc
     5521 gactctagag gatccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     5581 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     5641 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     5701 gtgccctggc ccaccctcgt gaccaccttc acctacggcg tgcagtgctt cagccgctac
     5761 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     5821 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     5881 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     5941 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     6001 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     6061 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     6121 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     6181 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactca cggcatggac
     6241 gagctgtaca agtaaagcgg ccgatcgttc aaacatttgg caataaagtt tcttaagatt
     6301 gaatcctgtt gccggtcttg cgatgattat catataattt ctgttgaatt acgttaagca
     6361 tgtaataatt aacatgtaat gcatgacgtt atttatgaga tgggttttta tgattagagt
     6421 cccgcaatta tacatttaat acgcgataga aaacaaaata tagcgcgcaa actaggataa
     6481 attatcgcgc gcggtgtcat ctatgttact agatccgatg ataagctgtc aaacatgaga
     6541 attcgtaatc atggtcatag ctgtttc
//
