LOCUS       sequence-271438-        3773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for generating in situ hybridization probe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3773)
  AUTHORS   Toriba T, Tokunaga H, Nagasawa K, Nie F, Yoshida A, Kyozuka J
  TITLE     Suppression of Leaf Blade Development by BLADE-ON-PETIOLE Orthologs 
            Is a Common Strategy for Underground Rhizome Growth.
  JOURNAL   Curr Biol. 2020 Feb 3;30(3):509-516.e3. doi: 
            10.1016/j.cub.2019.11.055. Epub 2020 Jan 16.
  PUBMED    31956025
REFERENCE   2  (bases 1 to 3773)
  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"; doi: 
            "10.1016/j.cub.2019.11.055"; journalName: "Curr Biol"; date: 
            "2020-02-3- 3"; volume: "30"; issue: "3"; pages: "509-516.e3"
COMMENT     Sequence Label: pGEM_HcBOP1binsitu
FEATURES             Location/Qualifiers
     source          1..3773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      50..59
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      54..63
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(915..933)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(916..933)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(951..967)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(951..967)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(964..986)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    975..991
                     /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(999..1005,1006..1023,1024..1029))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1044..1065
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1182..1199)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1353..1941)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1433..1452)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2112..2903,2904..2972))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2735..2754
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2973..3077)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3155..3610)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3292..3313)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3504..3523
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3736..3758
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3750..3767
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3751..3767
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattgggc ccgacgtcgc atgctcccgg ccgccatggc
       61 ggccgcggga attcgattca ttcacctttt cttatggatt gcgcttctgt atagcatttc
      121 gagactgcgt acgtgaagga gcaggatgag ctagtattca tgggaatata ttggcgggtc
      181 acagccatcc tgggattcat gccgattatt cgtgctcacc cttctgctgt tggtctcagg
      241 actacctcca ttgccatcca tcttgcaccc caattgattg agatacacca ttcttgagtc
      301 aaggttaagg ttcacaattg gcggctcagg gttcctcgag gcgtaaatgg cagtcggact
      361 ggcatcggcc ccactttcct cccgagaaag gaccatcgca gcggactgga cgagctcgag
      421 gcagagtcgg agcttgttgg gttcgacgtg ggccagacca gggatagcgc ctttgaagag
      481 gaaatcggac gtgagggtac ggaggatgtc gagcggggtc actccgtccg ccgtccggac
      541 tttagggtcg gcgtggtggt cgagcaggac ggctaccatg tccggacaga ccatttcagc
      601 ggctaggtgg agtggagtct tccccaccag accggcaggg caattaacgt cggctgcgcc
      661 aagctcgaga agcgccttca ccacctcccg gctgcagtac tggacggagt agtggagggc
      721 gagggcgtcg tcaaggttca ggccttctcc catgaccatc agcttgacga gctccacatc
      781 cgacgagtcg agagcccgcc gcatgcggcg aattttgttg tcgtccagct ctccaatcac
      841 tagtgaattc gcggccgcct gcaggtcgac catatgggag agctcccaac gcgttggatg
      901 catagcttga gtattctata gtgtcaccta aatagcttgg cgtaatcatg gtcatagctg
      961 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     1021 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     1081 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     1141 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     1201 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1261 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1321 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1381 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1441 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1501 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1561 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1621 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1681 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1741 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1801 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1861 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1921 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1981 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2041 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2101 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2161 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2221 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2281 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2341 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2401 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2461 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2521 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2581 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2641 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2701 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2761 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2821 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2881 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2941 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     3001 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     3061 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgatgcggt gtgaaatacc
     3121 gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta
     3181 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3241 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3301 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3361 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3421 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3481 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3541 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3601 cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     3661 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     3721 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttg
//
