LOCUS       sequence-466745-        5120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  G264A Cat-RNA U1367, Catalytically dead ribozyme control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5120)
  AUTHORS   Kalvapalle PB, Staubus A, Dysart MJ, Gambill L, Reyes Gamas K, Lu 
            LC, Silberg JJ, Stadler LB, Chappell J
  TITLE     Information storage across a microbial community using universal RNA
            barcoding.
  JOURNAL   Nat Biotechnol. 2025 Mar 18. doi: 10.1038/s41587-025-02593-0.
  PUBMED    40102641
REFERENCE   2  (bases 1 to 5120)
  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 
            Biotechnol. 2025 Mar 18. doi: 10.1038/s41587-025-02593-0."
COMMENT     Sequence Label: pRAM29
FEATURES             Location/Qualifiers
     source          1..5120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      424..1195
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             2470..2481
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    2528..2549
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    2579..2606
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     protein_bind    2637..2664
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     primer_bind     3573..3593
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    3706..3726
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(3864..4658)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3975..3994)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4585..4604
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     terminator      complement(4832..4863)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(join(4881..5120,1..423))
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
ORIGIN
        1 gccatgcccc aacagcgatt cgtcctggtc gtcgtgatca accaccacgg cggcactgaa
       61 caccgacagg cgcaactggt cgcggggctg gccccacgcc acgcggtcat tgaccacgta
      121 ggccgacacg gtgccggggc cgttgagctt cacgacggag atccagcgct cggccaccaa
      181 gtccttgact gcgtattgga ccgtccgcaa agaacgtccg atgagcttgg aaagtgtttt
      241 ctggctgacc accacggcgt tctggtggcc catctgcgcc acgaggtgat gcagcagcat
      301 tgccgccgtg ggtttcctcg caataagccc ggcccacgcc tcatgcgctt tgcgttccgt
      361 ttgcacccag tgaccgggct tgttcttggc ttgaatgccg atttctctgg actgcgtggc
      421 catgcttatc tccatgcggt aggggtgccg cacggttgcg gcaccatgcg caatcagctg
      481 caacttttcg gcagcgcgac aacaattatg cgttgcgtaa aagtggcagt caattacaga
      541 ttttctttaa cctacgcaat gagctattgc ggggggtgcc gcaatgagct gttgcgtacc
      601 cccctttttt aagttgttga tttttaagtc tttagcattt cgccctatat ctagttcttt
      661 ggtgcccaaa gaagggcacc cctgcggggt tcccccacgc cttcggcgcg gctccccctc
      721 cggcaaaaag tggcccctcc ggggcttgtt gatcgactgc gcggccttcg gccttgccca
      781 aggtggcgct gcccccttgg aacccccgca ctcgccgccg tgaggctcgg ggggcaggcg
      841 ggcgggcttc gcccttcgac tgcccccact cgcataggct tgggtcgttc caggcgcgtc
      901 aaggccaagc cgctgcgcgg tcgctgcgcg agccttgacc cgccttccac ttggtgtcca
      961 accggcaagc gaagcgcgca ggccgcaggc cggaggcttt tccccagaga aaattaaaaa
     1021 aattgatggg gcaaggccgc aggccgcgca gttggagccg gtgggtatgt ggtcgaaggc
     1081 tgggtagccg gtgggcaatc cctgtggtca agctcgtggg caggcgcagc ctgtccatca
     1141 gcttgtccag cagggttgtc cacgggccga gcgaagcgag ccagccggtg gccgctcgcg
     1201 gccatcgtcc acatatccac gggctggcaa gggagcgcag cgaccgcgca gggcgaagcc
     1261 cggagagcaa gcccgtaggg cgccgcagcc gccgtaggcg gtcacgactt tgcgaagcaa
     1321 agtctagtga gtatactcaa gcattgagtg gcccgccgga ggcaccgcct tgcgctgccc
     1381 ccgtcgagcc ggttggacac caaaagggag gggcaggcat ggcggcatac gcgatcatgc
     1441 gatgcaagaa gctggcgaaa atgggcaacg tggcggccag tctcaagcac gcctaccgcg
     1501 agcgcgagac tcccaacgct gacgccagca ggacgccaga gaacgagcac tgggcggcca
     1561 gcagcaccga tgaagcgatg ggccgactgc gcgagttgct gccagagaag cggcgcaagg
     1621 acgctgtgtt ggcggtcgag tacgtcatga cggccagccc ggaatggtgg aagtcggcca
     1681 gccaagaaca gcaggcggcg ttcttcgaga aggcgcacaa gtggctggcg gacaagtacg
     1741 gggcggatcg catcgtgacg gccagcatcc accgtgacga aaccagcccg cacatgaccg
     1801 cgttcgtggt gccgctgacg caggacggca ggctgtcggc caaggagttc atcggcaaca
     1861 aagcgcagat gacccgcgac cagaccacgt ttgcggcagc tgtggccgat ctagggctgc
     1921 aacggggcat cgagggcagc aaggcacgtc acacgcgcat tcaggcgttc tacgaggccc
     1981 tggagcggcc accagtgggc cacgtcacca tcagcccgca agcggtcgag ccacgcgcct
     2041 atgcaccgca gggattggcc gaaaagctgg gaatctcaaa gcgcgttgaa acgccggaag
     2101 ccgtggccga ccggctgaca aaagcggttc ggcaggggta tgagcctgcc ctacaggccg
     2161 ccgcaggagc gcgtgagatg cgcaagaagg ccgatcaagc ccaagagaca gcccgagatc
     2221 ttcgggagcg cctgaagccc gttctggacg ccctggggcc gttgaatcgg gatatgcagg
     2281 ccaaggccgc cgcgatcatc aaggccgtgg gcgaaaagct gctgacggaa cagcgggaag
     2341 tccagcgcca gaaacaggcc cagcgccagc aggaacgcgg gcgcgcacat ttccccgaaa
     2401 agtgccacct gggctgataa acggaaaaaa aaaccccgcc cctgacaggg cggggttttt
     2461 tttccaatta ttgaaggccg ctaacgcggc ctttttttgt ttctggtctg ccttaatcaa
     2521 tgactaagaa ttcgcggccg cttctagagc gtctcacgtt gctagccctg caggcttcaa
     2581 caaacagaca atctggtctg tttgtattat ggaaaatttt tctgtataat agattcaaca
     2641 aacagacaat ctggtctgtt tgtattatcg accaacccac tcccatggtg tgacgggcgg
     2701 tgtgtacaag gcccgggaac gtgttcacaa aagttatcag gcatgcacct ggtagctagt
     2761 ctttaaacca atagattgca tcggtttaaa aggcaagacc gtcaaattgc gggaaagggg
     2821 tcaacagccg ttcagtacca agtctcaggg gaaactttga gatggccttg caaagggtat
     2881 ggtaataagc tgacggacat ggtcctaacc acgcagccaa gtcctaagtc aacagatctt
     2941 ctgttgatat ggatgcagtt cacaaactaa atgtcggtcg gggaagatgt attcttctca
     3001 taagatatag tcggacctct ccttaatggg agctagcgga tgaagtgatg caacactgga
     3061 gccgctggga actaatttgt atgcgaaagt atattgatta gttttggagt actcgatggt
     3121 gttcaatgct tttcccgtta tccggatcac atgaaacggc atgacttttt caagagtgcc
     3181 atgcccgaag gttatgtaca ggaacgcact atatctttca aagatgacgg gacctacaag
     3241 acgcgtgctg aagtcaagtt tgaaggtgat acccttgtta atcgtatcga gttaaagggt
     3301 attgatttta aagaagatgg aaacattctt ggacacaaac tcgagtacaa ctttaactca
     3361 cacaatgtat acatcacggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt
     3421 cgccacaacg ttgaagatgg ttccgttcaa ctagcagacc attatcaaca aaatactcca
     3481 attggcgatg gccctgtcct tttaccagac aaccattacc tgtcgacaca atctgtcctt
     3541 tcgaaagatc ccaacgaaaa gcgtgaccac atggtccttc ttgagtttgt aactgctgct
     3601 gggattacac atggcatgga tgagctctac aaatggccca attattgaag gcctccctaa
     3661 cggggggcct ttttttgttt ctggtctgcc gctgcgtttg agacgtacta gtagcggccg
     3721 ctgcagttaa tcactgatta actcggtacc aaattccaga aaagaggcct cccgaaaggg
     3781 gggccttttt tcgttttggt ccttttgtta tcaataaaaa aggggagcgg tttcccgctc
     3841 cccttattgt tcgtctacaa ttattagaag aactcgtcaa gaaggcgata gaaggcgatg
     3901 cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg
     3961 ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca
     4021 cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc
     4081 aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg tgccttgagc
     4141 ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg
     4201 acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg
     4261 aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat
     4321 actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat
     4381 agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc
     4441 gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac
     4501 aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca
     4561 tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg
     4621 gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc
     4681 tcttgatcag atcttgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag
     4741 tttactttgc agggcttccc aaccttacca gagggcgccc cagctggcaa ttccggttcg
     4801 cttgctgtcc ataagagtcc tgttgagtaa tagtcaaaag cctcaggtcg gaggcttttg
     4861 actttctgct tacccaatta ctaccggcgc ggcagcgtga cccgtgtcgg cggctccaac
     4921 ggctcgccat cgtccagaaa acacggctca tcgggcatcg gcaggcgctg ctgcccgcgc
     4981 cgttcccatt cctccgtttc ggtcaaggct ggcaggtctg gttccatgcc cggaatgccg
     5041 ggctggctgg gcggctcctc gccggggccg gtcggtagtt gctgctcgcc cggatacagg
     5101 gtcgggatgc ggcgcaggtc
//
