LOCUS       sequence-467175-        5120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBBR1 origin cat-RNA U1376.
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: pRAM24
FEATURES             Location/Qualifiers
     source          1..5120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1135..1906
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1907..2569
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     terminator      2587..2618
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             2792..3586
                     /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(2846..2865)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3456..3475
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    3724..3744
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(3857..3877)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     protein_bind    4786..4813
                     /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    4844..4871
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     misc_feature    4901..4922
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(4969..4980)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 tccgtcagca gcttttcgcc cacggccttg atgatcgcgg cggccttggc ctgcatatcc
       61 cgattcaacg gccccagggc gtccagaacg ggcttcaggc gctcccgaag atctcgggct
      121 gtctcttggg cttgatcggc cttcttgcgc atctcacgcg ctcctgcggc ggcctgtagg
      181 gcaggctcat acccctgccg aaccgctttt gtcagccggt cggccacggc ttccggcgtt
      241 tcaacgcgct ttgagattcc cagcttttcg gccaatccct gcggtgcata ggcgcgtggc
      301 tcgaccgctt gcgggctgat ggtgacgtgg cccactggtg gccgctccag ggcctcgtag
      361 aacgcctgaa tgcgcgtgtg acgtgccttg ctgccctcga tgccccgttg cagccctaga
      421 tcggccacag ctgccgcaaa cgtggtctgg tcgcgggtca tctgcgcttt gttgccgatg
      481 aactccttgg ccgacagcct gccgtcctgc gtcagcggca ccacgaacgc ggtcatgtgc
      541 gggctggttt cgtcacggtg gatgctggcc gtcacgatgc gatccgcccc gtacttgtcc
      601 gccagccact tgtgcgcctt ctcgaagaac gccgcctgct gttcttggct ggccgacttc
      661 caccattccg ggctggccgt catgacgtac tcgaccgcca acacagcgtc cttgcgccgc
      721 ttctctggca gcaactcgcg cagtcggccc atcgcttcat cggtgctgct ggccgcccag
      781 tgctcgttct ctggcgtcct gctggcgtca gcgttgggag tctcgcgctc gcggtaggcg
      841 tgcttgagac tggccgccac gttgcccatt ttcgccagct tcttgcatcg catgatcgcg
      901 tatgccgcca tgcctgcccc tcccttttgg tgtccaaccg gctcgacggg ggcagcgcaa
      961 ggcggtgcct ccggcgggcc actcaatgct tgagtatact cactagactt tgcttcgcaa
     1021 agtcgtgacc gcctacggcg gctgcggcgc cctacgggct tgctctccgg gcttcgccct
     1081 gcgcggtcgc tgcgctccct tgccagcccg tggatatgtg gacgatggcc gcgagcggcc
     1141 accggctggc tcgcttcgct cggcccgtgg acaaccctgc tggacaagct gatggacagg
     1201 ctgcgcctgc ccacgagctt gaccacaggg attgcccacc ggctacccag ccttcgacca
     1261 catacccacc ggctccaact gcgcggcctg cggccttgcc ccatcaattt ttttaatttt
     1321 ctctggggaa aagcctccgg cctgcggcct gcgcgcttcg cttgccggtt ggacaccaag
     1381 tggaaggcgg gtcaaggctc gcgcagcgac cgcgcagcgg cttggccttg acgcgcctgg
     1441 aacgacccaa gcctatgcga gtgggggcag tcgaagggcg aagcccgccc gcctgccccc
     1501 cgagcctcac ggcggcgagt gcgggggttc caagggggca gcgccacctt gggcaaggcc
     1561 gaaggccgcg cagtcgatca acaagccccg gaggggccac tttttgccgg agggggagcc
     1621 gcgccgaagg cgtgggggaa ccccgcaggg gtgcccttct ttgggcacca aagaactaga
     1681 tatagggcga aatgcgaaag acttaaaaat caacaactta aaaaaggggg gtacgcaaca
     1741 gctcattgcg gcaccccccg caatagctca ttgcgtaggt taaagaaaat ctgtaattga
     1801 ctgccacttt tacgcaacgc ataattgttg tcgcgctgcc gaaaagttgc agctgattgc
     1861 gcatggtgcc gcaaccgtgc ggcaccccta ccgcatggag ataagcatgg ccacgcagtc
     1921 cagagaaatc ggcattcaag ccaagaacaa gcccggtcac tgggtgcaaa cggaacgcaa
     1981 agcgcatgag gcgtgggccg ggcttattgc gaggaaaccc acggcggcaa tgctgctgca
     2041 tcacctcgtg gcgcagatgg gccaccagaa cgccgtggtg gtcagccaga aaacactttc
     2101 caagctcatc ggacgttctt tgcggacggt ccaatacgca gtcaaggact tggtggccga
     2161 gcgctggatc tccgtcgtga agctcaacgg ccccggcacc gtgtcggcct acgtggtcaa
     2221 tgaccgcgtg gcgtggggcc agccccgcga ccagttgcgc ctgtcggtgt tcagtgccgc
     2281 cgtggtggtt gatcacgacg accaggacga atcgctgttg gggcatggcg acctgcgccg
     2341 catcccgacc ctgtatccgg gcgagcagca actaccgacc ggccccggcg aggagccgcc
     2401 cagccagccc ggcattccgg gcatggaacc agacctgcca gccttgaccg aaacggagga
     2461 atgggaacgg cgcgggcagc agcgcctgcc gatgcccgat gagccgtgtt ttctggacga
     2521 tggcgagccg ttggagccgc cgacacgggt cacgctgccg cgccggtagt aattgggtaa
     2581 gcagaaagtc aaaagcctcc gaccggaggc ttttgactat tactcaacag gactcttatg
     2641 gacagcaagc gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg
     2701 caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagatc
     2761 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     2821 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     2881 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     2941 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     3001 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3061 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3121 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3181 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3241 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     3301 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     3361 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     3421 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     3481 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     3541 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctaataat tgtagacgaa
     3601 caataagggg agcgggaaac cgctcccctt ttttattgat aacaaaagga ccaaaacgaa
     3661 aaaaggcccc cctttcggga ggcctctttt ctggaatttg gtaccgagtt aatcagtgat
     3721 taactgcagc ggccgctact agtacgtctc aaacgcagcg gcagaccaga aacaaaaaaa
     3781 ggccccccgt tagggaggcc ttcaataatt gggccatttg tagagctcat ccatgccatg
     3841 tgtaatccca gcagcagtta caaactcaag aaggaccatg tggtcacgct tttcgttggg
     3901 atctttcgaa aggacagatt gtgtcgacag gtaatggttg tctggtaaaa ggacagggcc
     3961 atcgccaatt ggagtatttt gttgataatg gtctgctagt tgaacggaac catcttcaac
     4021 gttgtggcga attttgaagt tagctttgat tccattcttt tgtttgtctg ccgtgatgta
     4081 tacattgtgt gagttaaagt tgtactcgag tttgtgtcca agaatgtttc catcttcttt
     4141 aaaatcaata ccctttaact cgatacgatt aacaagggta tcaccttcaa acttgacttc
     4201 agcacgcgtc ttgtaggtcc cgtcatcttt gaaagatata gtgcgttcct gtacataacc
     4261 ttcgggcatg gcactcttga aaaagtcatg ccgtttcatg tgatccggat aacgggaaaa
     4321 gcattgaaca ccatcgagta ctccaaaact aatcaatata ctttcgcata caaattagtt
     4381 cccagcggct ccagtgttgc atcacttcat ccgctagctc ccattaagga gaggtccgac
     4441 tatatcttat gagaagaata catcttcccc gaccgacatt tagtctgtga actgcatcca
     4501 tatcaacaga agatctgttg acttaggact tggctgcgtg gttaggacca tgtccgtcag
     4561 cttattacca taccctttgc aaggccatct caaagtttcc cctgagactt ggtactgaac
     4621 ggctgttgac ccctttcccg caatttgacg gtcttgcctt ttaaaccgat gcaatctatt
     4681 ggtttaaaga ctagctacca ggtgcatgcc tgataacttt tgtgaacacg ttcccgggcc
     4741 ttgtacacac cgcccgtcac accatgggag tgggttggtc gataatacaa acagaccaga
     4801 ttgtctgttt gttgaatcta ttatacagaa aaattttcca taatacaaac agaccagatt
     4861 gtctgtttgt tgaagcctgc agggctagca acgtgagacg ctctagaagc ggccgcgaat
     4921 tcttagtcat tgattaaggc agaccagaaa caaaaaaagg ccgcgttagc ggccttcaat
     4981 aattggaaaa aaaaccccgc cctgtcaggg gcggggtttt tttttccgtt tatcagccca
     5041 ggtggcactt ttcggggaaa tgtgcgcgcc cgcgttcctg ctggcgctgg gcctgtttct
     5101 ggcgctggac ttcccgctgt
//
