LOCUS       sequence-479403-        2765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Node A carrying sgRNA-1 that can be used for the assembly of 
            pEND-Y11.
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-Y
FEATURES             Location/Qualifiers
     source          1..2765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(811..838)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(930..1016)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(1025..1072)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1068..1086
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    complement(1448..1467)
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     protein_bind    complement(1574..1593)
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     rep_origin      1910..2498
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2399..2418
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2564..2655
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2656..2765,1..682)
                     /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"
ORIGIN
        1 agtggatgac ggcctgaagc cacacagtga tattgatttg ctggttacgg tgaccgtaag
       61 gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt cggcttcccc
      121 tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg acgacatcat
      181 tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc gcaatgacat
      241 tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct tgctgacaaa
      301 agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct ttgatccggt
      361 tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga actcgccgcc
      421 cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc
      481 agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc
      541 ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac aagaagaaga
      601 tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga aaggcgagat
      661 caccaaggta gtcggcaaat aatgtctaac aattcgttca agccgagggg ccgcaagatc
      721 cggccacgat gacccggtcg tcggttcagg gcagggtcgt taaatagccg cttatgtcta
      781 ttgctggttt accggtagat ctgaaacgca aaaaggccat ccgtcaggat ggccttctgc
      841 ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc
      901 ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac
      961 aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc
     1021 ctggcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct agttattgct
     1081 cagcggtggc agcaaagctt attttctaca gtctcagttt cagggatttc gttcacttta
     1141 ccttcttact caataacaac ctatctacta ccaggcacca gttctcgcaa cagtgtatca
     1201 attaccaact tactgaaatc acagaactgg gtgggactcg gcagagaaaa ctcaatcgga
     1261 gactacttta ctatctatta ttcgttacag gtcagaagaa ttgagaagag aaaagaaaac
     1321 cgccgatcct gtccaccgca ttactgcaag gtagtggaca agaccggcgg tcttaagttt
     1381 tttggctgaa ccatggtaac ggagaagccc tatcaactgt ttattgctcg agcgggcgtc
     1441 ccagcgactg cctatacggc agtgaacgca ccgactcggt gccacttttt caagttgata
     1501 acggactagc cttatttcaa cttgctatgc tgtttccagc atagctctga aaccactaga
     1561 gtctagcttg agactgccta tacggcagtg aacccggacc gcaggctgga gctcggacgg
     1621 taaagtttcc agaatctatc ctatgaattc aggtggcact tttcggggaa atgtgcgagt
     1681 cactgcatcc taggagcaac ccagtcagct ccttccggtg ggcgcggggc atgactaaca
     1741 tgagaattac aacttatatc gtatggggct gacttcaggt gctacatttg aagagataaa
     1801 ttgcactgaa atctagagcg gttcagtaga aaatcatgac caaaatccct taacgtgagt
     1861 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1921 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1981 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2041 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2101 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2161 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2221 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2281 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2341 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2401 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2461 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggccccgc
     2521 cgcagtctca cgcccggagc gtagcgaccg agtgagctag ctatttgttt atttttctaa
     2581 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2641 tgaaaaggga agagtatgag ggaagcggtg atcgccgaag tatcgactca actatcagag
     2701 gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc
     2761 tccgc
//
