LOCUS       sequence-348328-        2638 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 2638)
  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 2638)
  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: pTAG1
FEATURES             Location/Qualifiers
     source          1..2638
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    141..162
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(177..182,183..200,201..207)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    215..231
                     /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     220..242
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     239..255
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     239..255
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             251..688
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGATSRLR
                     FRARNSRDHEVPRHEPSGHKPVRFVSCNASSVCAHATGPEP"
     primer_bind     274..294
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        276..294
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(341..360)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(342..360)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(370..387)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(370..386)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(379..401)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             877..1668
                     /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      2009..2597
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2498..2517
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggtctcaaat gaacaaagca ccagtggtct agtggtagaa tagtaccctg ccacggtaca
       61 gacccgggtt cgattcccgg ctggtgcatg agacgagatc tagacgaccg gtggaaagcg
      121 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
      181 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
      241 ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa
      301 aagctggagg acctcgaggg ggggcccggt acccaattcg ccctatagtg agtcgtatta
      361 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
      421 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
      481 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
      541 cggcgccact agtcgtctcc gttttagagc tagaaatagc cgagaccacg aagtgccgag
      601 gcacgaaccc agtggacata agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg
      661 cgctcacgca actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg
      721 gttttcatgg cttgttatga ctgttttttt ggggtacagt ctatgcctcg ggcatccaag
      781 cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg
      841 cagcagggca gtcgccctaa aacaaagtta aacatcatgg gggaagcggt gatcgccgaa
      901 gtatcgactc aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg
      961 ctggccgtac atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagcgatatt
     1021 gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac
     1081 gaccttttgg aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc
     1141 accattgttg tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa
     1201 tttggagaat ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac
     1261 attgatctgg ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca
     1321 gcggcggagg aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa
     1381 accttaacgc tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt
     1441 acgttgtccc gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct
     1501 gccgactggg caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga
     1561 caggcttatc ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa
     1621 tttgtccatt acgtgaaagg cgagatcacc aaggtagtcg gcaaataatg tctagctaga
     1681 aattcgttca agccgacgcc gcttcgcggc gcggcttaac tcaagcgtta gatgcactaa
     1741 gcacataatt gctcacagcc aaactatcag gtcaagtctg cttttattat ttttaagcgt
     1801 gcataataag ccctacacaa attgggagat atatcatgct gtcagaccaa gtttactcat
     1861 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     1921 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     1981 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2041 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2101 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     2161 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2221 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2281 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2341 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2401 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2461 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2521 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2581 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg cactctgt
//
