LOCUS       sequence-146146-        2893 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSEVA331 with Biobrick multiple cloning site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2893)
  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 2893)
  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: pSEVA331Bb
FEATURES             Location/Qualifiers
     source          1..2893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..21
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      26..120
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        168..270
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             271..930
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(337..356)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     oriT            1079..1187
                     /note="incP origin of transfer"
     CDS             complement(1198..1860)
                     /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      1861..2632
                     /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(2747..2833)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2848..2870
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    2872..2893
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 tactagtagc ggccgctgca gcttggactc ctgttgatag atccagtaat gacctcagaa
       61 ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat tggtgagaat
      121 ccaggggtcc ccaataatta cgatttaaat tggcgaaaat gagacgttga tcggcacgta
      181 agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta
      241 tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc
      301 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa
      361 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa
      421 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat
      481 ccggaatttc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct
      541 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac
      601 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac
      661 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg
      721 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
      781 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
      841 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
      901 tactgcgatg agtggcaggg cggggcgtaa tttgactttt gtccttttcc gctgcataac
      961 cctgcttcgg ggtcattata gcgatttttt cggtatatcc atcctttttc gcacgatata
     1021 caggattttg ccaaagggtt cgtgtagact ttccttggtg tatccaacgg cgtcagccgg
     1081 gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc cttcttcact gtcccttatt
     1141 cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg ctggccgtag gccggcccta
     1201 ccggcgcggc agcgttaccc gtgtcggcgg ctccaacggc tcgccatcgt ccagaaaaca
     1261 cggctcatcg ggcatcggca ggcgctgctg cccgcgccgt tcccattcct ccgtttcggt
     1321 caaggctggc aggtctggtt ccatgcccgg aatgccgggc tggctgggcg gctcctcgcc
     1381 ggggccggtc ggtagttgct gctcgcccgg atacagggtc gggatgcggc gcaggtcgcc
     1441 atgccccaac agcgattcgt cctggtcgtc gtgatcaacc accacggcgg cactgaacac
     1501 cgacaggcgc aactggtcgc ggggctggcc ccacgccacg cggtcattga ccacgtaggc
     1561 cgacacggtg ccggggccgt tgagcttcac gacggagatc cagcgctcgg ccaccaagtc
     1621 cttgactgcg tattggaccg tccgcaaaga acgtccgatg agcttggaaa gtgtcttctg
     1681 gctgaccacc acggcgttct ggtggcccat ctgcgccacg aggtgatgca gcagcattgc
     1741 cgccgtgggt ttcctcgcaa taagcccggc ccacgcctca tgcgctttgc gttccgtttg
     1801 cacccagtga ccgggcttgt tcttggcttg aatgccgatt tctctggact gcgtggccat
     1861 gcttatctcc atgcggtagg ggtgccgcac ggttgcggca ccatgcgcaa tcagctgcaa
     1921 cttttcggca gcgcgacaac aattatgcgt tgcgtaaaag tggcagtcaa ttacagattt
     1981 tctttaacct acgcaatgag ctattgcggg gggtgccgca atgagctgtt gcgtaccccc
     2041 cttttttaag ttgttgattt ttaagtcttt cgcatttcgc cctatatcta gttctttggt
     2101 gcccaaagaa gggcacccct gcggggttcc cccacgcctt cggcgcggct ccccctccgg
     2161 caaaaagtgg cccctccggg gcttgttgat cgactgcgcg gccttcggcc ttgcccaagg
     2221 tggcgctgcc cccttggaac ccccgcactc gccgccgtga ggctcggggg gcaggcgggc
     2281 gggcttcgcc cttcgactgc ccccactcgc ataggcttgg gtcgttccag gcgcgtcaag
     2341 gccaagccgc tgcgcggtcg ctgcgcgagc cttgacccgc cttccacttg gtgtccaacc
     2401 ggcaagcgaa gcgcgcaggc cgcaggccgg aggcttttcc ccagagaaaa ttaaaaaaat
     2461 tgatggggca aggccgcagg ccgcgcagtt ggagccggtg ggtatgtggt cgaaggctgg
     2521 gtagccggtg ggcaatccct gtggtcaagc tcgtgggcag gcgcagcctg tccatcagct
     2581 tgtccagcag ggttgtccac gggccgagcg aagcgagcca gccggtggcc gctcgcggcc
     2641 atcgtccaca tatccacggg ctggcaaggg agcgcagcga ccgcgcaggg cgaagcccgg
     2701 agagcaagcc cgtagggggg gcgcgcccag ctgtctaggg cggcggattt gtcctactca
     2761 ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc gactgagcct
     2821 ttcgttttat ttgatgcctt taattaaagc ggataacaat ttcacacagg agaattcgcg
     2881 gccgcttcta gag
//
