LOCUS       sequence-232858-        3205 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression esgRNA in rice protoplasts.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3205)
  AUTHORS   Li C, Zong Y, Wang Y, Jin S, Zhang D, Song Q, Zhang R, Gao C
  TITLE     Expanded base editing in rice and wheat using a Cas9-adenosine 
            deaminase fusion.
  JOURNAL   Genome Biol. 2018 May 29;19(1):59. doi: 10.1186/s13059-018-1443-z.
  PUBMED    29807545
REFERENCE   2  (bases 1 to 3205)
  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"; journalName: "Genome 
            Biol."; date: "2018-05-29"; volume: "19(1):59. doi"; pages: " 
            10.1186/s13059-018-1443-z"
COMMENT     Sequence Label: pOsU3-esgRNA
FEATURES             Location/Qualifiers
     source          1..3205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     73..95
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     87..104
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     88..104
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        168..547
                     /label=OsU3 promoter
                     /note="Oryza sativa (rice) snRNA U3 promoter"
     terminator      655..674
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     primer_bind     complement(693..709)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(693..709)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(706..728)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    717..733
                     /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(741..747,748..765,766..771))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    786..807
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(924..941)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1095..1683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1175..1194)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1854..2645,2646..2714))
                     /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     2477..2496
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2715..2819)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2887..2905
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2943..2965)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3065..3084
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
       61 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattc agtaattcat
      121 ccaggtcacc aagttctagg attttcagaa ctgcaactta ttttatcaag gaatctttaa
      181 acatacgaac agatcactta aagttcttct gaagcaactt aaagttatca ggcatgcatg
      241 gatcttggag gaatcagatg tgcagtcagg gaccatagca caagacaggc gtcttctact
      301 ggtgctacca gcaaatgctg gaagccggga acactgggta cgttggaaac cacgtgatgt
      361 gaagaagtaa gataaactgt aggagaaaag catttcgtag tgggccatga agcctttcag
      421 gacatgtatt gcagtatggg ccggcccatt acgcaattgg acgacaacaa agactagtat
      481 tagtaccacc tcggctatcc acatagatca aagctgattt aaaagagttg tgcagatgat
      541 ccgtggcgtg agaccctgca gggtctctgt ttaagagcta tgctggaaac agcatagcaa
      601 gtttaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
      661 tgttttttat gtctaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
      721 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
      781 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
      841 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
      901 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
      961 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1021 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1081 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1141 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1201 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     1261 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     1321 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1381 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1441 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1501 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     1561 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1621 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1681 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     1741 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     1801 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     1861 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     1921 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     1981 gcaatgatac cgcgagaacc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     2041 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     2101 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     2161 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     2221 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     2281 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     2341 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     2401 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     2461 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     2521 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     2581 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     2641 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     2701 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     2761 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     2821 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     2881 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     2941 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     3001 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     3061 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     3121 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     3181 tgggaagggc gatcggtgcg ggcct
//
