LOCUS       sequence-271434-        4489 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 4489)
  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 4489)
  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_ZmaBOP2binsitu
FEATURES             Location/Qualifiers
     source          1..4489
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1654..1672)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1655..1672)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1690..1706)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1690..1706)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1703..1725)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1714..1730
                     /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(1738..1744,1745..1762,1763..1768))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1783..1804
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1921..1938)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2092..2680)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2172..2191)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2851..3642,3643..3711))
                     /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     3474..3493
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3712..3816)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3894..4349)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4031..4052)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4243..4262
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(4475..4489,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4489,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg ggcccaagcc
       61 attggggaag tacaagttgg atgggctccc tcggccgccg ttccctttcc tcgtgtcgcc
      121 ggccactcca gcgtccatct cctggccctg gtacatgagc gttgagttca tcgtcaggct
      181 caccaagctg tcgtcggcgt cgttccttgc gaaatttccg ccgtcgcttc ccccggcgtc
      241 gcaaccaccg gcaggcggtg cgccctcgtc gcgcgtggtc accaacaccg cggactgcac
      301 gagctcaagg cacagcctga gcttgttggg ctcgatgtgc gtgagccccg gcacggcgcc
      361 cttgaagagg aactccgagg tgaggccgcg gagcacgtcg agcggggtga ctccgtcgag
      421 cgtgctggcg ttcgggtccg cgtggtggtc gaggaggacg gacaccaggt ccggcgagac
      481 catctcggct gccaggtgga gcggcgtctt ccccgtcggc cccgcgcggg agttgacgtc
      541 ggccgcgccg agctccagca gcgccttgac cacgtcgcgg ttgcagtgct ggaccgcgta
      601 gtgcacggcg agcgcgtcgt cgaggtccag accctcgctc atgaccatca gcttgacgag
      661 ctcgatgtcg gccgcgtcca gcgcccgccg catgcgccgg atcttgtggt cgcggtcagc
      721 ggcggcggag gccggagcgt tgatggggag gtagtactgc gtcgagaacg gcgagcgcgt
      781 cgtgctcctc gccgcggccg caaccggtga cttggcgcgg atctcctcga tcttggccac
      841 cacgtcgatg gggaggtgct tggagaggag gtcggccgag aggcctgagc gtgcgacgag
      901 gtgggggcag gtgtcccaga gctcctgcat ctcgaacttg cgcgacgcca atagcacctt
      961 catcacgtcg tcgacggacg cctccttcac catgctctcc agctgcttct ggaccaagag
     1021 cgcgagctgc tcgacgccga aggagcgcgc ggcggcgagc gcgtcgaggg cgaggtcgac
     1081 ggcggcgccg caccgggtgt gccagcaccc gcgggcgccg cacccgggca gcggcccgct
     1141 cttgggcgcg gccaccgacg cctggccgct gtagaggaac tgcagcacga gcacgagcac
     1201 ctcgtaccgg atcgagctga cggggatgac cagctccggc gtggcgccag ccgagccacg
     1261 ggacgccgga ggcagctggt ggttgggatc gaggccgcag aagagcttgc ggaagaagag
     1321 gctgcgcgcg gcgaggacgc agcggtgggc gtggacgagg cggccctcgg cgctgaacgc
     1381 gacgtcgctg aacgcctggc cgttgatgag caggttgagg tagtcgagcg acagcgactt
     1441 gaccgagtcc tcggagctca tgatcgatat ctcatgtgcg gcagcagcag cacccggccg
     1501 gtaaaggcga ccgatggatc tatcgatcga gctactagga ccgaatcgcg ctggctggtc
     1561 cagcttttcg atcggaggag aaggaattcg cggccgcctg caggtcgacc atatgggaga
     1621 gctcccaacg cgttggatgc atagcttgag tattctatag tgtcacctaa atagcttggc
     1681 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1741 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1801 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1861 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1921 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1981 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2041 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2101 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2161 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2221 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2281 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2341 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2401 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2461 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2521 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2581 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2641 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2701 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2761 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2821 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2881 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2941 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3001 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3061 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3121 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3181 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3241 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3301 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3361 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3421 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3481 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3541 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3601 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3661 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3721 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3781 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3841 tgatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt
     3901 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     3961 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     4021 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     4081 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     4141 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     4201 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     4261 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     4321 cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg
     4381 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     4441 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgtt
//
