LOCUS       sequence-466009-        4873 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pColE1 origin Cat-RNA U1376, Multiplexing ribozyme splicing 
            reactions for the purpose of comparing host range of origins of 
            replications.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4873)
  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 4873)
  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: pRAM28
FEATURES             Location/Qualifiers
     source          1..4873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             95..286
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(280..302)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    388..528
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     443..462
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(543..560)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(714..1302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(794..813)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2520..2531
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    2578..2599
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    2629..2656
                     /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    2687..2714
                     /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     3628..3648
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    3761..3781
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(3919..4713)
                     /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(4030..4049)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4640..4659
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc
       61 catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct
      121 taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct
      181 ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta
      241 ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
      301 ggtgacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
      361 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
      421 agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
      481 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
      541 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      601 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      661 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      721 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      781 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      841 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      901 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      961 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     1021 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     1081 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     1141 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     1201 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1261 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1321 tctacgggac ggccgcagcc gccgtaggcg gtcacgactt tgcgaagcaa agtctagtga
     1381 gtatactcaa gcattgagtg gcccgccgga ggcaccgcct tgcgctgccc ccgtcgagcc
     1441 ggttggacac caaaagggag gggcaggcat ggcggcatac gcgatcatgc gatgcaagaa
     1501 gctggcgaaa atgggcaacg tggcggccag tctcaagcac gcctaccgcg agcgcgagac
     1561 tcccaacgct gacgccagca ggacgccaga gaacgagcac tgggcggcca gcagcaccga
     1621 tgaagcgatg ggccgactgc gcgagttgct gccagagaag cggcgcaagg acgctgtgtt
     1681 ggcggtcgag tacgtcatga cggccagccc ggaatggtgg aagtcggcca gccaagaaca
     1741 gcaggcggcg ttcttcgaga aggcgcacaa gtggctggcg gacaagtacg gggcggatcg
     1801 catcgtgacg gccagcatcc accgtgacga aaccagcccg cacatgaccg cgttcgtggt
     1861 gccgctgacg caggacggca ggctgtcggc caaggagttc atcggcaaca aagcgcagat
     1921 gacccgcgac cagaccacgt ttgcggcagc tgtggccgat ctagggctgc aacggggcat
     1981 cgagggcagc aaggcacgtc acacgcgcat tcaggcgttc tacgaggccc tggagcggcc
     2041 accagtgggc cacgtcacca tcagcccgca agcggtcgag ccacgcgcct atgcaccgca
     2101 gggattggcc gaaaagctgg gaatctcaaa gcgcgttgaa acgccggaag ccgtggccga
     2161 ccggctgaca aaagcggttc ggcaggggta tgagcctgcc ctacaggccg ccgcaggagc
     2221 gcgtgagatg cgcaagaagg ccgatcaagc ccaagagaca gcccgagatc ttcgggagcg
     2281 cctgaagccc gttctggacg ccctggggcc gttgaatcgg gatatgcagg ccaaggccgc
     2341 cgcgatcatc aaggccgtgg gcgaaaagct gctgacggaa cagcgggaag tccagcgcca
     2401 gaaacaggcc cagcgccagc aggaacgcgg gcgcgcacat ttccccgaaa agtgccacct
     2461 gggctgataa acggaaaaaa aaaccccgcc cctgacaggg cggggttttt tttccaatta
     2521 ttgaaggccg ctaacgcggc ctttttttgt ttctggtctg ccttaatcaa tgactaagaa
     2581 ttcgcggccg cttctagagc gtctcacgtt gctagccctg caggcttcaa caaacagaca
     2641 atctggtctg tttgtattat ggaaaatttt tctgtataat agattcaaca aacagacaat
     2701 ctggtctgtt tgtattatcg accaacccac tcccatggtg tgacgggcgg tgtgtacaag
     2761 gcccgggaac gtgttcacaa aagttatcag gcatgcacct ggtagctagt ctttaaacca
     2821 atagattgca tcggtttaaa aggcaagacc gtcaaattgc gggaaagggg tcaacagccg
     2881 ttcagtacca agtctcaggg gaaactttga gatggccttg caaagggtat ggtaataagc
     2941 tgacggacat ggtcctaacc acgcagccaa gtcctaagtc aacagatctt ctgttgatat
     3001 ggatgcagtt cacagactaa atgtcggtcg gggaagatgt attcttctca taagatatag
     3061 tcggacctct ccttaatggg agctagcgga tgaagtgatg caacactgga gccgctggga
     3121 actaatttgt atgcgaaagt atattgatta gttttggagt actcgatggt gttcaatgct
     3181 tttcccgagt ggttatccgg atcacatgaa acggcatgac tttttcaaga gtgccatgcc
     3241 cgaaggttat gtacaggaac gcactatatc tttcaaagat gacgggacct acaagacgcg
     3301 tgctgaagtc aagtttgaag gtgataccct tgttaatcgt atcgagttaa agggtattga
     3361 ttttaaagaa gatggaaaca ttcttggaca caaactcgag tacaacttta actcacacaa
     3421 tgtatacatc acggcagaca aacaaaagaa tggaatcaaa gctaacttca aaattcgcca
     3481 caacgttgaa gatggttccg ttcaactagc agaccattat caacaaaata ctccaattgg
     3541 cgatggccct gtccttttac cagacaacca ttacctgtcg acacaatctg tcctttcgaa
     3601 agatcccaac gaaaagcgtg accacatggt ccttcttgag tttgtaactg ctgctgggat
     3661 tacacatggc atggatgagc tctacaaatg gcccaattat tgaaggcctc cctaacgggg
     3721 ggcctttttt tgtttctggt ctgccgctgc gtttgagacg tactagtagc ggccgctgca
     3781 gttaatcact gattaactcg gtaccaaatt ccagaaaaga ggcctcccga aaggggggcc
     3841 ttttttcgtt ttggtccttt tgttatcaat aaaaaagggg agcggtttcc cgctcccctt
     3901 attgttcgtc tacaattatt agaagaactc gtcaagaagg cgatagaagg cgatgcgctg
     3961 cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag
     4021 ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag
     4081 ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca
     4141 ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc
     4201 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
     4261 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
     4321 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
     4381 ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag
     4441 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
     4501 ggccagccac gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc
     4561 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
     4621 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
     4681 agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg
     4741 atcagatctt gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac
     4801 tttgcagggc ttcccaacct taccagaggg cgccccagct ggcaattccg gttcgcttgc
     4861 tgtccataac caa
//
