LOCUS       sequence-319674-        3218 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tRNA and scaffold for the assembly of GBoligomers for position [4-5]
            of a polycistronic tRNA-gRNA..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3218)
  AUTHORS   Vazquez-Vilar M, Garcia-Carpintero V, Selma S, Bernabe-Orts JM, 
            Sanchez-Vicente J, Salazar-Sarasua B, Ressa A, de Paola C, Ajenjo M,
            Quintela JC, Fernandez-Del-Carmen A, Granell A, Orzaez D
  TITLE     The GB4.0 Platform, an All-In-One Tool for CRISPR/Cas-Based 
            Multiplex Genome Engineering in Plants.
  JOURNAL   Front Plant Sci. 2021 Jul 1;12:689937. doi: 
            10.3389/fpls.2021.689937. eCollection 2021.
  PUBMED    34276739
REFERENCE   2  (bases 1 to 3218)
  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: "Front Plant
            Sci."; date: "2021-07-1"; volume: "12:689937. doi"; pages: " 
            10.3389/fpls.2021.689937. eCollection 2021"
COMMENT     Sequence Label: pVD1 Multiplexing Edit E4 (GB2241)
FEATURES             Location/Qualifiers
     source          1..3218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     30..49
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(268..372)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(450..905)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(587..608)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     799..818
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1031..1053
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1045..1062
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1046..1062
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1069..1088
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1069..1087
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1118..1127
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1122..1131
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1428..1446)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1429..1446)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1464..1480)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1464..1480)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1477..1499)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1488..1504
                     /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(1512..1518,1519..1536,1537..1542))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1557..1578
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1695..1712)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1866..2454)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1946..1965)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2625..3218,1..198,199..267))
                     /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"
ORIGIN
        1 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
       61 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
      121 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
      181 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
      241 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
      301 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
      361 aggggttccg cgcacatttc cccgaaaagt gccacctgat gcggtgtgaa ataccgcaca
      421 gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt
      481 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
      541 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
      601 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
      661 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
      721 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
      781 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
      841 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
      901 cgcgtccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      961 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     1021 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgta atacgactca
     1081 ctatagggcg aattgggccc gacgtcgcat gctcccggcc gccatggcgg ccgcgggaat
     1141 tcgattcccg tctcgctcac tcggtgccac tttttcaagt tgataacgga ctagccttat
     1201 tttaacttgc tatttctagc tctaaaactg agacggcgcg cgccgtctct gcaccagccg
     1261 ggaatcgaac ccgggtctgt accgtggcag ggtactattc taccactaga ccactggtgc
     1321 tttgttgcac cgactcgagc gagacgggat cactagtgaa ttcgcggccg cctgcaggtc
     1381 gaccatatgg gagagctccc aacgcgttgg atgcatagct tgagtattct atagtgtcac
     1441 ctaaatagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1501 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     1561 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1621 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1681 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1741 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     1801 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1861 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1921 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1981 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2041 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2101 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2161 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2221 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2281 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2341 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2401 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2461 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2521 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2581 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2641 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2701 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2761 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     2821 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     2881 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     2941 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3001 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3061 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3121 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3181 tcaaccaagt cattctgaga atagtgtatg cggcgacc
//
