LOCUS       sequence-146144-        4562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An inducible small RNA with E.coli Hfq targeting K. rhaeticus UGPase
            mRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4562)
  AUTHORS   Florea M, Hagemann H, Santosa G, Abbott J, Micklem CN, 
            Spencer-Milnes X, de Arroyo Garcia L, Paschou D, Lazenbatt C, Kong 
            D, Chughtai H, Jensen K, Freemont PS, Kitney R, Reeve B, Ellis T
  TITLE     Engineering control of bacterial cellulose production using a 
            genetic toolkit and a new cellulose-producing strain.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Jun 14;113(24):E3431-40. doi: 
            10.1073/pnas.1522985113. Epub 2016 May 31.
  PUBMED    27247386
REFERENCE   2  (bases 1 to 4562)
  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: 
            "10.1073/pnas.1522985113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-06-14- 14"; volume: "113"; issue: "24"; pages: 
            "E3431-40"
COMMENT     Sequence Label: J-sRNA-331Bb
FEATURES             Location/Qualifiers
     source          1..4562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             46..57
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      861..932
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      948..975
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             1163..1174
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      1695..1789
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        1837..1939
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1940..2599
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2006..2025)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     oriT            2748..2856
                     /note="incP origin of transfer"
     CDS             complement(2867..3529)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      3530..4301
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      complement(4416..4502)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4517..4539
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    4541..4562
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 tggtgcaaaa cctttcgcgg tatggcatga tagcgcctac tagagaaaga ggagaaatac
       61 tagatgaaaa acataaatgc cgacgacaca tacagaataa ttaataaaat taaagcttgt
      121 agaagcaata atgatattaa tcaatgctta tctgatatga ctaaaatggt acattgtgaa
      181 tattatttac tcgcgatcat ttatcctcat tctatggtta aatctgatat ttcaatccta
      241 gataattacc ctaaaaaatg gaggcaatat tatgatgacg ctaatttaat aaaatatgat
      301 cctatagtag attattctaa ctccaatcat tcaccaatta attggaatat atttgaaaac
      361 aatgctgtaa ataaaaaatc tccaaatgta attaaagaag cgaaaacatc aggtcttatc
      421 actgggttta gtttccctat tcatacggct aacaatggct tcggaatgct tagttttgca
      481 cattcagaaa aagacaacta tatagatagt ttatttttac atgcgtgtat gaacatacca
      541 ttaattgttc cttctctagt tgataattat cgaaaaataa atatagcaaa taataaatca
      601 aacaacgatt taaccaaaag agaaaaagaa tgtttagcgt gggcatgcga aggaaaaagc
      661 tcttgggata tttcaaaaat attaggttgc agtgagcgta ctgtcacttt ccatttaacc
      721 aatgcgcaaa tgaaactcaa tacaacaaac cgctgccaaa gtatttctaa agcaatttta
      781 acaggagcaa ttgattgccc atactttaaa aattaataac actgatagtg ctagtgtaga
      841 tcactactag agccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
      901 gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc tcaccttcgg
      961 gtgggccttt ctgcgtttat atactagaga cctgtaggat cgtacaggtt tacgcaagaa
     1021 aatggtttgt tatagtcgaa tggctttttt aaggggctta atcactttct gttgggccat
     1081 tgcattgcca ctgattttcc aacatataaa aagacaagcc cgaacagtcg tccgggcttt
     1141 ttttctcgag ctcgagccaa ttattgaagg ccgctaacgc ggcctttttt tgtttctggt
     1201 ctccctacta gtaacctgta ggatcgtaca ggtttacgca agaaaatggt ttgttatagt
     1261 cgaataaaat gcatgacacg caaatactac aaggaggaat tctatatggc taaggggcaa
     1321 tctttacaag atccgttcct gaacgcactg cgtcgggaac gtgttccagt ttctatttat
     1381 ttggtgaatg gtattaagct gcaagggcaa atcgagtctt ttgatcagtt cgtgatcctg
     1441 ttgaaaaaca cggtcagcca gatggtttac aagcacgcga tttctactgt tgtcccgtct
     1501 cgcccggttt ctcatcacag taacaacgcc ggtggcggta ccagcagtaa ctaccatcat
     1561 ggtagcagcg cgcagaatac ttccgcgcaa caggacagcg aagaaaccga ataatactag
     1621 ctcggtacca aattccagaa aagagacgct ttcgagcgtc ttttttcgtt ttggtccgcg
     1681 gccgctgcag cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga
     1741 tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc caggggtccc
     1801 caataattac gatttaaatt ggcgaaaatg agacgttgat cggcacgtaa gaggttccaa
     1861 ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc
     1921 aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc
     1981 ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa
     2041 ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa
     2101 gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaatttcg
     2161 tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt
     2221 tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg
     2281 gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt
     2341 ccctaaaggg tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac
     2401 cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg
     2461 caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc
     2521 cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga
     2581 gtggcagggc ggggcgtaat ttgacttttg tccttttccg ctgcataacc ctgcttcggg
     2641 gtcattatag cgattttttc ggtatatcca tcctttttcg cacgatatac aggattttgc
     2701 caaagggttc gtgtagactt tccttggtgt atccaacggc gtcagccggg caggataggt
     2761 gaagtaggcc cacccgcgag cgggtgttcc ttcttcactg tcccttattc gcacctggcg
     2821 gtgctcaacg ggaatcctgc tctgcgaggc tggccgtagg ccggccctac cggcgcggca
     2881 gcgttacccg tgtcggcggc tccaacggct cgccatcgtc cagaaaacac ggctcatcgg
     2941 gcatcggcag gcgctgctgc ccgcgccgtt cccattcctc cgtttcggtc aaggctggca
     3001 ggtctggttc catgcccgga atgccgggct ggctgggcgg ctcctcgccg gggccggtcg
     3061 gtagttgctg ctcgcccgga tacagggtcg ggatgcggcg caggtcgcca tgccccaaca
     3121 gcgattcgtc ctggtcgtcg tgatcaacca ccacggcggc actgaacacc gacaggcgca
     3181 actggtcgcg gggctggccc cacgccacgc ggtcattgac cacgtaggcc gacacggtgc
     3241 cggggccgtt gagcttcacg acggagatcc agcgctcggc caccaagtcc ttgactgcgt
     3301 attggaccgt ccgcaaagaa cgtccgatga gcttggaaag tgtcttctgg ctgaccacca
     3361 cggcgttctg gtggcccatc tgcgccacga ggtgatgcag cagcattgcc gccgtgggtt
     3421 tcctcgcaat aagcccggcc cacgcctcat gcgctttgcg ttccgtttgc acccagtgac
     3481 cgggcttgtt cttggcttga atgccgattt ctctggactg cgtggccatg cttatctcca
     3541 tgcggtaggg gtgccgcacg gttgcggcac catgcgcaat cagctgcaac ttttcggcag
     3601 cgcgacaaca attatgcgtt gcgtaaaagt ggcagtcaat tacagatttt ctttaaccta
     3661 cgcaatgagc tattgcgggg ggtgccgcaa tgagctgttg cgtacccccc ttttttaagt
     3721 tgttgatttt taagtctttc gcatttcgcc ctatatctag ttctttggtg cccaaagaag
     3781 ggcacccctg cggggttccc ccacgccttc ggcgcggctc cccctccggc aaaaagtggc
     3841 ccctccgggg cttgttgatc gactgcgcgg ccttcggcct tgcccaaggt ggcgctgccc
     3901 ccttggaacc cccgcactcg ccgccgtgag gctcgggggg caggcgggcg ggcttcgccc
     3961 ttcgactgcc cccactcgca taggcttggg tcgttccagg cgcgtcaagg ccaagccgct
     4021 gcgcggtcgc tgcgcgagcc ttgacccgcc ttccacttgg tgtccaaccg gcaagcgaag
     4081 cgcgcaggcc gcaggccgga ggcttttccc cagagaaaat taaaaaaatt gatggggcaa
     4141 ggccgcaggc cgcgcagttg gagccggtgg gtatgtggtc gaaggctggg tagccggtgg
     4201 gcaatccctg tggtcaagct cgtgggcagg cgcagcctgt ccatcagctt gtccagcagg
     4261 gttgtccacg ggccgagcga agcgagccag ccggtggccg ctcgcggcca tcgtccacat
     4321 atccacgggc tggcaaggga gcgcagcgac cgcgcagggc gaagcccgga gagcaagccc
     4381 gtaggggggg cgcgcccagc tgtctagggc ggcggatttg tcctactcag gagagcgttc
     4441 accgacaaac aacagataaa acgaaaggcc cagtctttcg actgagcctt tcgttttatt
     4501 tgatgccttt aattaaagcg gataacaatt tcacacagga gaattcgcgg ccgcttctag
     4561 ag
//
