LOCUS       sequence-348322-        2668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  First sgRNA in a tRNA separated array of sgRNAs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2668)
  AUTHORS   Cody JP, Maher MF, Nasti RA, Starker CG, Chamness JC, Voytas DF
  TITLE     Direct delivery and fast-treated Agrobacterium co-culture 
            (Fast-TrACC) plant transformation methods for Nicotiana benthamiana.
  JOURNAL   Nat Protoc. 2022 Oct 17. pii: 10.1038/s41596-022-00749-9. doi: 
            10.1038/s41596-022-00749-9.
  PUBMED    36253612
REFERENCE   2  (bases 1 to 2668)
  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: "Nat Protoc.
            2022 Oct 17. pii: 10.1038/s41596-022-00749-9. doi: 
            10.1038/s41596-022-00749-9."
COMMENT     Sequence Label: pTAG3
FEATURES             Location/Qualifiers
     source          1..2668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    181..202
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(217..222,223..240,241..247)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    255..271
                     /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)."
     primer_bind     260..282
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     279..295
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     279..295
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             291..668
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGATSRLR
                     FRAETTKCRGTNPVDISLFGS"
     primer_bind     314..334
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        316..334
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(381..400)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(382..400)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(410..427)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(410..426)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(419..441)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             907..1698
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      2039..2627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2528..2547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggtctcaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg caacaaagca
       61 ccagtggtct agtggtagaa tagtaccctg ccacggtaca gacccgggtt cgattcccgg
      121 ctggtgcatg agacgagatc tagacgaccg gtggaaagcg ggcagtgagc gcaacgcaat
      181 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
      241 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      301 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagg acctcgaggg
      361 ggggcccggt acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg
      421 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
      481 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
      541 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgccact agtcgtctcc
      601 gttttagagc cgagaccacg aagtgccgag gcacgaaccc agtggacata agcctgttcg
      661 gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc
      721 gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt
      781 ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt
      841 ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta
      901 aacatcatgg gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc
      961 gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg
     1021 gatggcggcc tgaagccaca cagcgatatt gatttgctgg ttacggtgac cgtaaggctt
     1081 gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga
     1141 gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg
     1201 tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt
     1261 gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca
     1321 agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct
     1381 gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac
     1441 tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta
     1501 accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc
     1561 cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc
     1621 ttggcctcgc gcgcagatca gttggaagaa tttgtccatt acgtgaaagg cgagatcacc
     1681 aaggtagtcg gcaaataatg tctagctaga aattcgttca agccgacgcc gcttcgcggc
     1741 gcggcttaac tcaagcgtta gatgcactaa gcacataatt gctcacagcc aaactatcag
     1801 gtcaagtctg cttttattat ttttaagcgt gcataataag ccctacacaa attgggagat
     1861 atatcatgct gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     1921 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     1981 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2041 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2101 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2161 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     2221 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2281 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2341 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2401 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2461 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2521 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2581 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2641 cggccttttt acggttcctg cactctgt
//
