LOCUS       sequence-479360-        2782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Node C carrying sgRNA-1 that can be used for the assembly of 
            pEND-11Y or pEND-ZY.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2782)
  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 2782)
  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: pNC-Y
FEATURES             Location/Qualifiers
     source          1..2782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        239..273
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     promoter        239..273
                     /label=BBa_J23107
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.36"
     promoter        239..273
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     protein_bind    417..436
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     protein_bind    543..562
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     primer_bind     complement(765..783)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      779..826
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      835..921
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1013..1040
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(1169..1960)
                     /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(1961..2052)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2118..2706)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2198..2217)
                     /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 gaagtattgg taatcgttga aaactcagtc gaccagcctg cggtccggtt
      241 tacggctagc tcagtcctag gtattatgct agctcgctgg gacgcccgtg aacagttgct
      301 ctgattgaaa ccacgattca gcctgcggtc cggccaagta cttagtacac actgattcgc
      361 tgggacgccc ggtagtgctt atcagaccca atactgttga acagcctgcg gtccgggttc
      421 actgccgtat aggcagtctc aagctagact ctagtggttt cagagctatg ctggaaacag
      481 catagcaagt tgaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
      541 gcgttcactg ccgtataggc agtcgctggg acgcccggac gtcctattac actcgtcgtt
      601 ggaaactgaa gatgcggccg cggaaacaca gaaaaaagcc cgcacctgac agtgcgggct
      661 ttttttttcg accaaaggta gcgaacgacg agtcactgtt gaggataaat actttctcta
      721 ctaggcgcct gttacacagg tcctcagcgg cgcgcctgct gccaccgctg agcaataact
      781 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgccag gcatcaaata
      841 aaacgaaagg ctcagtcgga agactgggcc tttcgtttta tctgttgttt gtcggtgaac
      901 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgtgaa gcaacggccc
      961 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaactaa gcagaaggcc
     1021 atcctgacgg atggcctttt tgcgtttcag atctaccggt aaaccagcaa tagacataag
     1081 cggctattta acgaccctgc cctgaaccga cgaccgggtc atcgtggccg gatcttgcgg
     1141 cccctcggct tgaacgaatt gttagacatt atttgccgac taccttggtg atctcgcctt
     1201 tcacgtagtg gacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttctt
     1261 gtccaagata agcctgtcta gcttcaagta tgacgggctg atactgggcc ggcaggcgct
     1321 ccattgccca gtcggcagcg acatccttcg gcgcgatttt gccggttact gcgctgtacc
     1381 aaatgcggga caacgtaagc actacatttc gctcatcgcc agcccagtcg ggcggcgagt
     1441 tccatagcgt taaggtttca tttagcgcct caaatagatc ctgttcagga accggatcaa
     1501 agagttcctc cgccgctgga cctaccaagg caacgctatg ttctcttgct tttgtcagca
     1561 agatagccag atcaatgtcg atcgtggctg gctcgaagat acctgcaaga atgtcattgc
     1621 gctgccattc tccaaattgc agttcgcgct tagctggata acgccacgga atgatgtcgt
     1681 cgtgcacaac aatggtgact tctacagcgc ggagaatctc gctctctcca ggggaagccg
     1741 aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc cttacggtca
     1801 ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc gtcatccact gcggagccgt
     1861 acaaatgtac ggccagcaac gtcggttcga gatggcgctc gatgacgcca actacctctg
     1921 atagttgagt cgatacttcg gcgatcaccg cttccctcat actcttcctt tttcaatatt
     1981 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     2041 aaaataaaca aatagctagc tcactcggtc gctacgctcc gggcgtgaga ctgcggcggg
     2101 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2161 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2221 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2281 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2341 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2401 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2461 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2521 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2581 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2641 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2701 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2761 cgttaaggga ttttggtcat ga
//
