LOCUS       sequence-495026-        2765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Node A carrying sgRNA-2 that can be used for the assembly of 
            pEND-Z11 or pEND-ZY.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2765)
  AUTHORS   Nevot G, Guell M, Santos-Moreno J
  TITLE     Critical Analysis of Preprints and Inquiry-Based Lessons Improve the
            Synthetic Biology Learning Experience.
  JOURNAL   ACS Synth Biol. 2025 Aug 15;14(8):2878-2884. doi: 
            10.1021/acssynbio.5c00014.
  PUBMED    40814254
REFERENCE   2  (bases 1 to 2765)
  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: "10.1021/acssynbio";
            journalName: "ACS Synth Biol"; date: "2025-08-15- 15"; volume: "14";
            issue: "8"; pages: "2878-2884"
COMMENT     Sequence Label: pNA-Z
FEATURES             Location/Qualifiers
     source          1..2765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    241..260
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     protein_bind    367..386
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     primer_bind     complement(748..766)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      762..809
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      818..904
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      996..1023
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(1152..1943)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDDGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(1944..2035)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2101..2689)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2181..2200)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttctactg aaccgctcta gatttcagtg caatttatct cttcaaatgt agcacctgaa
       61 gtcagcccca tacgatataa gttgtaattc tcatgttagt catgccccgc gcccaccgga
      121 aggagctgac tgggttgctc ctaggatgca gtgactcgca catttccccg aaaagtgcca
      181 cctgaattca taggatagat tctggaaact ttaccgtccg agctccagcc tgcggtccgg
      241 gttcactgcc gtataggcag atcagtgtgt actaagtact gtttcagagc tatgctggaa
      301 acagcatagc aagttgaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
      361 cggtgcgttc actgccgtat aggcagtcgc tgggacgccc gctcgagcaa taaacagttg
      421 atagggcttc tccgttacca tggttcagcc aaaaaactta agaccgccgg tcttgtccac
      481 taccttgcag taatgcggtg gacaggatcg gcggttttct tttctcttct caattcttct
      541 gacctgtaac gaataataga tagtaaagta gtctccgatt gagttttctc tgccgagtcc
      601 cacccagttc tgtgatttca gtaagttggt aattgataca ctgttgcgag aactggtgcc
      661 tggtagtaga taggttgtta ttgagtaaga aggtaaagtg aacgaaatcc ctgaaactga
      721 gactgtagaa aataagcttt gctgccaccg ctgagcaata actagcataa ccccttgggg
      781 cctctaaacg ggtcttgagg ggttttttgc caggcatcaa ataaaacgaa aggctcagtc
      841 ggaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
      901 aaatccgccg ggagcggatt tgaacgttgt gaagcaacgg cccggagggt ggcgggcagg
      961 acgcccgcca taaactgcca ggcatcaaac taagcagaag gccatcctga cggatggcct
     1021 ttttgcgttt cagatctacc ggtaaaccag caatagacat aagcggctat ttaacgaccc
     1081 tgccctgaac cgacgaccgg gtcatcgtgg ccggatcttg cggcccctcg gcttgaacga
     1141 attgttagac attatttgcc gactaccttg gtgatctcgc ctttcacgta gtggacaaat
     1201 tcttccaact gatctgcgcg cgaggccaag cgatcttctt cttgtccaag ataagcctgt
     1261 ctagcttcaa gtatgacggg ctgatactgg gccggcaggc gctccattgc ccagtcggca
     1321 gcgacatcct tcggcgcgat tttgccggtt actgcgctgt accaaatgcg ggacaacgta
     1381 agcactacat ttcgctcatc gccagcccag tcgggcggcg agttccatag cgttaaggtt
     1441 tcatttagcg cctcaaatag atcctgttca ggaaccggat caaagagttc ctccgccgct
     1501 ggacctacca aggcaacgct atgttctctt gcttttgtca gcaagatagc cagatcaatg
     1561 tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat tgcgctgcca ttctccaaat
     1621 tgcagttcgc gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg
     1681 acttctacag cgcggagaat ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg
     1741 ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac cagcaaatca
     1801 atatcactgt gtggcttcag gccgtcatcc actgcggagc cgtacaaatg tacggccagc
     1861 aacgtcggtt cgagatggcg ctcgatgacg ccaactacct ctgatagttg agtcgatact
     1921 tcggcgatca ccgcttccct catactcttc ccttttcaat attattgaag catttatcag
     1981 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaatagct
     2041 agctcactcg gtcgctacgc tccgggcgtg agactgcggc ggggccgcgt tgctggcgtt
     2101 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2161 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2221 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2281 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2341 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2401 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2461 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2521 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2581 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2641 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2701 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2761 catga
//
