LOCUS       40924_2737        5631 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Moss transformation vector p3xMyc aphIV DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5631)
  AUTHORS   Nishiyama T, Cheng C, Tamada Y, Kurata T, Hasebe M.
  TITLE     Moss transformation vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5631)
  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 5631)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5631)
  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            "Unpublished"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..5631
                     /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..730
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     CDS             746..775
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             782..811
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_recomb     843..876
                     /label=loxP
                     /note="loxP"
     protein_bind    843..876
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     terminator      881..1132
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        1215..1562
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             1582..2604
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    2652..2828
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     protein_bind    complement(3317..3350)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(3443..3461)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3482..3498)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3506..3522)
                     /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(3530..3560)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3575..3596)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3884..4472)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4646..5503)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5504..5608)
                     /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 gaacaaaaat taatctcaga
      721 agaagacttg aacggactcg acggtgaaca aaagttgatt tctgaagaag atttgaacgg
      781 tgaacaaaag ctaatctccg aggaagactt ggtgtacaag taagaattcc tgcagccccg
      841 aaataacttc gtatagcata cattatacga agttatattc ctcgttcaaa catttggcaa
      901 taaagtttct taagattgaa tcctgttgcc ggtcttgcga tgattatcat ataatttctg
      961 ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt tatgagatgg
     1021 gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa caaaatatag
     1081 cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactaga tccgatgata
     1141 agctgtcaaa catgagaatt gcccccctac tccaaaaatg tcaaagatac agtctcagaa
     1201 gaccaaaggg ctattgacac ttttcaacaa agggtatata atcgggaaga ctcctcggat
     1261 tccattgccc agctatctgt cacttcatcg aaaggacagt agaaaaggaa ggtggctcct
     1321 acaaatgcca tcattgcgat aaaggaaagg ctatcattca agatgcctct gccgacagtg
     1381 gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca
     1441 cgtcttcaaa gcaagtggat tgatgtgaca tctccactga cgtaagggat gacgcacaat
     1501 cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga
     1561 cagcccaagc aaggaggtga tatgaagaag cctgaactca ccgcgacgtc tgtcgagaag
     1621 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
     1681 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
     1741 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
     1801 attccggaag tgcttgacat tggagagttc agcgagagcc tgacctattg catctcccgc
     1861 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttcttcag
     1921 ccggtcgcgg aggctatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
     1981 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg
     2041 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
     2101 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
     2161 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg
     2221 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc
     2281 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat
     2341 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa
     2401 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc
     2461 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
     2521 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc
     2581 agcactcgtc ctagggcaaa ggaatagcgc tgaaatcacc agtctctctc tacaaatcta
     2641 tctctctcta ttttctccat aaataatgtg tgagtagttt cccgataagg gaaattaggg
     2701 ttcttatagg gtttcgctca tgtgttgagc atataagaaa cccttagtat gtatttgtat
     2761 ttgtaaaata cttctatcaa taaaatttct aattcctaaa accaaaatcc agtactaaaa
     2821 tccagatctc ctaaagtccc tatagatctt tgtcgtgaat ataaaccaga cacgagacga
     2881 ctaaacctgg agcccagacg ccgttcgaag ctagaagtac cgcttaggca ggaggccgtt
     2941 agggaaaaga tgctaaggca gggttggtta cgttgactcc cccgtaggtt tggtttaaat
     3001 atgatgaagt ggacggaagg aaggaggaag acaaggaagg ataaggttgc aggccctgtg
     3061 caaggtaaga agatggaaat ttgatagagg tacgctacta tacttatact atacgctaag
     3121 ggaatgcttg tatttatacc ctataccccc taataacccc ttatcaattt aagaaataat
     3181 ccgcataagc ccccgcttaa aaattggtat cagggccatg aataggtcta tgaccaaaac
     3241 tcaagaggat aaaacctcac caaaatacga aagagttctt aactctaaag ataaaagatc
     3301 tttcaagatc aaaactataa cttcgtatag catacattat acgaagttat ccctcacacc
     3361 ggtgacgggg atcgggggga tccacgtgcc cgggactagt tctagagcgg ccgccaccgc
     3421 ggtggagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     3481 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3541 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3601 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3661 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3721 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3781 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3841 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3901 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3961 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4021 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4081 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4141 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4201 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4261 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4321 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4381 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4441 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4501 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4561 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4621 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4681 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4741 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4801 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4861 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4921 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4981 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5041 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5101 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5161 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5221 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5281 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5341 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5401 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5461 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5521 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5581 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca c
//
