LOCUS       sequence-495027-        3604 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Node B carrying the sfGPF reporter that can be used for the assembly
            of pEND-11Y, pEND-Y11, pEND-Z11 or pEND-ZY.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 241 to 275)
  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 3604)
  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: pNB-Rep
FEATURES             Location/Qualifiers
     source          1..3604
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        241..275
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     protein_bind    419..438
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     RBS             443..465
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             473..1186
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1587..1605)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1601..1648
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1657..1743
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1835..1862
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(1991..2782)
                     /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(2783..2874)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2940..3528)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3020..3039)
                     /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 cctgaattcg aaatccctga aactgagact gtagaaaata agcttcagcc tgcggtccgg
      241 ttgacggcta gctcagtcct aggtacagtg ctagctcgct gggacgcccg agatagccgt
      301 tacacaggtg acacttattt cagcctgcgg tccggccaca ctagagtcta gcttgagatc
      361 gctgggacgc ccgtagaact ctgtgaggat aaagtctccc ttacagcctg cggtccgggt
      421 tcactgccgt ataggcagta attttgttta actttaagaa ggagatatac atatgcgtaa
      481 aggcgaagaa ctgtttaccg gtgtggttcc gattctggtg gaactggacg gcgatgttaa
      541 tggtcataaa ttcagtgttc gcggcgaagg tgaaggcgat gcgacgaacg gcaaactgac
      601 cctgaaattt atctgcacca cgggtaaact gccggtcccg tggccgacgc tggtgaccac
      661 gctgacctat ggcgttcaat gttttgcgcg ttacccggat cacatgaaac agcacgactt
      721 tttcaaatcg gccatgccgg aaggctatgt gcaggaacgt acgattagct ttaaagacga
      781 tggtacgtat aaaacccgcg cggaagtgaa attcgaaggc gataccctgg ttaaccgtat
      841 cgaactgaaa ggtatcgatt tcaaagaaga cggcaatatt ctgggtcata aactggaata
      901 taacttcaat tcccacaacg tgtacatcac cgcggataaa cagaaaaacg gcattaaagc
      961 caatttcaaa atccgccata atgtggaaga tggtagcgtt cagctggccg accactatca
     1021 gcaaaacacg ccgattggtg atggcccggt cctgctgccg gacaatcact acctgagtac
     1081 ccagtccgtg ctgtcaaaag atccgaacga aaaacgtgac cacatggtcc tgctggaatt
     1141 tgtgacggct gcgggtatca cccacggcat ggacgaactg tataaaatgt cccgccgtaa
     1201 tactgacgcc atcacaatcc acagcatcct ggattggatt gaagactaat aatcgctggg
     1261 acgcccggga tccaagagat ttctacacga ttgagcactg tctcctgcag gctcggtacc
     1321 aaattccaga aaagaggcct cccgaaaggg gggccttttt tcgttttggt cctaatagat
     1381 aaaggatagg tctggtagtg ttgttcgttc tcgcaggtaa atcaataata ctctgcagtt
     1441 ccgtagactt ttcagtggga cagggtagcg ataacagata gattgtaata agacacagta
     1501 ggtgctcgta gttgcgtgaa gagaaccgct caggaaatcc agtcagaagt attggtaatc
     1561 gttgaaaact cagtcgactg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     1621 ctctaaacgg gtcttgaggg gttttttgcc aggcatcaaa taaaacgaaa ggctcagtcg
     1681 gaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
     1741 aatccgccgg gagcggattt gaacgttgtg aagcaacggc ccggagggtg gcgggcagga
     1801 cgcccgccat aaactgccag gcatcaaact aagcagaagg ccatcctgac ggatggcctt
     1861 tttgcgtttc agatctaccg gtaaaccagc aatagacata agcggctatt taacgaccct
     1921 gccctgaacc gacgaccggg tcatcgtggc cggatcttgc ggcccctcgg cttgaacgaa
     1981 ttgttagaca ttatttgccg actaccttgg tgatctcgcc tttcacgtag tggacaaatt
     2041 cttccaactg atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc
     2101 tagcttcaag tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag
     2161 cgacatcctt cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa
     2221 gcactacatt tcgctcatcg ccagcccagt cgggcggcga gttccatagc gttaaggttt
     2281 catttagcgc ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg
     2341 gacctaccaa ggcaacgcta tgttctcttg cttttgtcag taagatagcc agatcaatgt
     2401 cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt gcgctgccat tctccaaatt
     2461 gcagttcgcg cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga
     2521 cttctacagc gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt
     2581 tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa
     2641 tatcactgtg tggcttcagg ccgtcatcca ctgcggagcc gtacaaatgt acggccagca
     2701 acgtcggttc gagatggcgc tcgatgacgc caactacctc tgatagttga gtcgatactt
     2761 cggcgatcac cgcttccctc atactcttcc tttttcaata ttattgaagc atttatcagg
     2821 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagcta
     2881 gctcactcgg tcgctacgct ccgggcgtga gactgcggcg gggccgcgtt gctggcgttt
     2941 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3001 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3061 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3121 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3181 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3241 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3301 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3361 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3421 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3481 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3541 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3601 atga
//
