LOCUS       40924_2732        5405 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Moss transformation vector p3xHA NPTII DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5405)
  AUTHORS   Nishiyama T, Cheng C, Tamada Y, Kurata T, Hasebe M.
  TITLE     Moss transformation vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5405)
  AUTHORS   Nishiyama T, Cheng C, Tamada Y, Kurata T, Hasebe M.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-NOV-2015) Contact:Yosuke Tamada National Institute for
            Basic Biology, Division of Evolutionary Biology; 38 Nishigonaka, 
            Myodaiji, Okazaki, Aichi 444-8585, Japan URL 
            :http://www.nibb.ac.jp/evodevo/
REFERENCE   3  (bases 1 to 5405)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5405)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-NOV-2015) Contact:Yosuke Tamada National Institute for Basic 
            Biology, Division of Evolutionary Biology; 38 Nishigonaka, Myodaiji,
            Okazaki, Aichi 444-8585, Japan URL :http://www.nibb.ac.jp/evodevo/"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             701..790
                     /codon_start=1
                     /label=3xHA
                     /note="three tandem HA epitope tags"
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     misc_recomb     822..855
                     /label=loxP
                     /note="loxP"
     protein_bind    822..855
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     terminator      860..1111
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        1194..1541
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             1562..2353
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    2426..2602
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     protein_bind    complement(3091..3124)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(3217..3235)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3256..3272)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3280..3296)
                     /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)."
     promoter        complement(3304..3334)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3349..3370)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3658..4246)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4420..5277)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5278..5382)
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgataa gcttgatatc tacccatacg atgttcctga
      721 ctatgcgggc tatccctatg acgtcccgga ctatgcagga tcctatccat atgacgttcc
      781 agattacgct gtgtacaagt aagaattcct gcagccccga aataacttcg tatagcatac
      841 attatacgaa gttatattcc tcgttcaaac atttggcaat aaagtttctt aagattgaat
      901 cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt taagcatgta
      961 ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat tagagtcccg
     1021 caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta ggataaatta
     1081 tcgcgcgcgg tgtcatctat gttactagat ccgatgataa gctgtcaaac atgagaattg
     1141 cccccctact ccaaaaatgt caaagataca gtctcagaag accaaagggc tattgacact
     1201 tttcaacaaa gggtatataa tcgggaagac tcctcggatt ccattgccca gctatctgtc
     1261 acttcatcga aaggacagta gaaaaggaag gtggctccta caaatgccat cattgcgata
     1321 aaggaaaggc tatcattcaa gatgcctctg ccgacagtgg tcccaaagat ggacccccac
     1381 ccacgaggag catcgtggaa aaagaagacg ttccaaccac gtcttcaaag caagtggatt
     1441 gatgtgacat ctccactgac gtaagggatg acgcacaatc ccactatcct tcgcaagacc
     1501 cttcctctat ataaggaagt tcatttcatt tggagaggac agcccaagca aggaggtgag
     1561 catgattgaa caagacggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     1621 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     1681 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     1741 ccaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     1801 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     1861 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     1921 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     1981 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     2041 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc ggatgcccga
     2101 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     2161 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     2221 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     2281 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     2341 tgacgagttc ttctgagcgg gactctgggg ttcggacggt acgctgaaat caccagtctc
     2401 tctctacaaa tctatctctc tctattttct ccataaataa tgtgtgagta gtttcccgat
     2461 aagggaaatt agggttctta tagggtttcg ctcatgtgtt gagcatataa gaaaccctta
     2521 gtatgtattt gtatttgtaa aatacttcta tcaataaaat ttctaattcc taaaaccaaa
     2581 atccagtact aaaatccaga tctcctaaag tccctataga tctttgtcgt gaatataaac
     2641 cagacacgag acgactaaac ctggagccca gacgccgttc gaagctagaa gtaccgctta
     2701 ggcaggaggc cgttagggaa aagatgctaa ggcagggttg gttacgttga ctcccccgta
     2761 ggtttggttt aaatatgatg aagtggacgg aaggaaggag gaagacaagg aaggataagg
     2821 ttgcaggccc tgtgcaaggt aagaagatgg aaatttgata gaggtacgct actatactta
     2881 tactatacgc taagggaatg cttgtattta taccctatac cccctaataa ccccttatca
     2941 atttaagaaa taatccgcat aagcccccgc ttaaaaattg gtatcagggc catgaatagg
     3001 tctatgacca aaactcaaga ggataaaacc tcaccaaaat acgaaagagt tcttaactct
     3061 aaagataaaa gatctttcaa gatcaaaact ataacttcgt atagcataca ttatacgaag
     3121 ttatccctca caccggtgac ggggatcggg gggatccacg tgcccgggac tagttctaga
     3181 gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt taattgcgcg
     3241 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3301 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3361 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3421 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3481 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3541 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3601 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3661 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3721 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3781 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3841 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3901 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3961 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4021 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4081 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4141 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4201 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4261 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4321 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4381 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4441 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4501 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4561 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4621 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4681 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4741 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4801 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4861 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4921 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4981 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5041 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5101 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5161 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5221 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5281 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5341 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5401 gccac
//
