LOCUS       sequence-364507-        9961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli - B. subtilis nicking vector pProUSER13F4C. Ap-BBR1; 
            Delta-sigF-insertion-site; Km; manR-PmanA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9961)
  AUTHORS   Falkenberg KB, Mol V, de la Maza Larrea AS, Pogrebnyakov I, Norholm 
            MHH, Nielsen AT, Jensen SI
  TITLE     The ProUSER2.0 Toolbox: Genetic Parts and Highly Customizable 
            Plasmids for Synthetic Biology in Bacillus subtilis.
  JOURNAL   ACS Synth Biol. 2021 Dec 17;10(12):3278-3289. doi: 
            10.1021/acssynbio.1c00130. Epub 2021 Nov 18.
  PUBMED    34793671
REFERENCE   2  (bases 1 to 9961)
  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: 
            "10.1021/acssynbio.1c00130"; journalName: "ACS Synth Biol"; date: 
            "2021-12-17- 17"; volume: "10"; issue: "12"; pages: "3278-3289"
COMMENT     Sequence Label: pProUSER13F4C
FEATURES             Location/Qualifiers
     source          1..9961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(27..113)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     128..150
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     terminator      4269..4363
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        6473..6564
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6565..6633,6634..7425)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      7429..7468
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            7615..7723
                     /note="incP origin of transfer"
     CDS             complement(7734..8396)
                     /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      8397..9168
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 gcgcgcccag ctgtctaggg cggcggattt gtcctactca ggagagcgtt caccgacaaa
       61 caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctt
      121 taattaaagc ggataacaat ttcacacagg aggccgatcg cgtcatagca aactcaaaga
      181 gtataaaaat cgcttttttc cggaagcttc ggtaaaaaac gaaacttttg tctctatgat
      241 tttgttttat aatgtaaacg gtttcttata tagtatactt atactatcaa tttgctcaag
      301 tagatactga caggaaggat agaaaaacag atggaataca taaatacaag acaaaaagag
      361 attttgtatc tgttgttatc tgaacctgat gactatttag ttgtgcaaga tttcgcagat
      421 agggtacagt gttctgaaaa aacaatccgg aatgatttga aagtgatcga ggattatctc
      481 aatgagcatt ctcatgcaca gcttattcga aaaccagggt tgggcgttta tttacatata
      541 gaagaacagg aaaggacctg gctaagccaa caattacata ctgaacattt ttcttctagg
      601 cagcggagcg ataaggaaag aatgctgcac attgcttatg acttattaat gaatccgaag
      661 cctgtttccg caaaagacat tgccgctcgg cattttgtca atcgatcttc cattaaaaag
      721 gatttatacg cagtagaaga gtggctgaag cgttttgatc tcacactggt ttctaggcag
      781 cggcttggat taaaggttga agggaacgaa agaaataaaa gaaaggcatt ggcaagaata
      841 tctgatttga ttcacaatac agcgtttacc agccaattta ttaaaagtaa gtttttacat
      901 tatgaagtag actttgtgac gaaagaaatc aaatcactac aaaaaaagca ttccctctat
      961 ttcacagacg aaacgtttga aagtttgttg ctgcatacat tgttaatggt ccgccgcatc
     1021 aaaatgaagc agccgatttc attgtcccca aaagaaatgg cggcagtaaa aaagaagaaa
     1081 gaatatcaat ggacatttgc ttgtttacag cggcttgaac cggtttttgc aattcgcttc
     1141 cctgaagaag aagccgtgta cttaacatta catatattgg gcggtaaggt tcgttatcca
     1201 ttgcagacag aagagaacct tgaaaatgct gtgcttccta aggttgtagg gcatttaatc
     1261 aatcgtgttt cagagctgaa aatgatggat tttcacaagg atcaggattt gatcaatggt
     1321 ttgaacattc atctcaatac ygtgctacaa cggctgagct acgatctttc tgtagcaaat
     1381 cctatgctca atgacatcaa aaagatgtac ccttatttgt tccacctcat cattgatgtt
     1441 ttggaagaca tcaatcaaac ttttgatctt catattcctg aggaagaggc agcatatttg
     1501 acattgcatt ttcaggcagc gatcgaacgg atgcagggca gcagcgagac acataaaaag
     1561 gcggttatcg tctgccacat ggggattggg atgtcacaat tgctgcggac aaaaattgaa
     1621 cggaaatatc accaaatcgc tgtaatggct tgtattgcaa aggccgattt aaaggattat
     1681 atcaaaaagc atgaggatat tgatcttgtc atatccacca ttgcgctgga gaatataacg
     1741 gttccccata ttgttgtgtc tccgctttta gagcctggcg aagagaagaa attaagtgcg
     1801 tttatacgtc agctgggcga atcacaccgc caaaaacaaa aaacgtttca gatgctgaac
     1861 aatacgactc cttttttggt ttttttgcag caggaggcgg agcatcgcta caagttaatc
     1921 gaacagctgg cgacagcttt gtttgagaaa ggctatgttg ataaggacta tgcggttcat
     1981 gcggtgatga gggaaaaaat gtcggctaca aatattgggt caggcattgc cattcctcat
     2041 gcaaacgcta aatttattaa acaatcagca atcgcgattg ccacattaaa ggagccgctt
     2101 gaatggggaa ataaaaaagt ttcgctcgtt tttatgctgg ctgtcaaaca cgaggatcaa
     2161 actatgacaa agcagctgtt tagcgagctt tcatatctta gcgagcagcc ggccttcgtc
     2221 cagaagctga cgaaagaaac caatgtcatg acatttttat ctcatttgga ttattaaaag
     2281 cagggattat tccttgcttt ttttgttata gggaaaaatg cctttattac cggaacctat
     2341 ggtaaaaaaa gcgattttaa tgagctgatt tcggtataca gttgagacaa gatcttatta
     2401 ttcacacttt aagctgaggc gatcgcaacg tcgtgactgg gaaaaccctg gcgttaccca
     2461 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     2521 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgct ttgcctggtt
     2581 tccggcacca gaagcggtgc cggaaagctg gctggagtgc gatcttcctg aggccgatac
     2641 tgtcgtcgtc ccctcaaact ggcagatgca cggttacgat gcgcccatct acaccaacgt
     2701 gacctatccc attacggtca atccgccgtt tgttcccacg gagaatccga cgggttgtta
     2761 ctcgctcaca tttaatgttg atgaaagctg gctacaggaa ggccagacgc gaattatttt
     2821 tgatggcgtt aactcggcgt ttcatctgtg gtgcaacggg cgctgggtcg gttacggcca
     2881 ggacagtcgt ttgccgtctg aatttgacct gagcgcattt ttacgcgccg gagaaaaccg
     2941 cctcgcggtg atggtgctgc gctggagtga cggcagttat ctggaagatc aggatatgtg
     3001 gcggatgagc ggcattttcc gtgacgtctc gttgctgcat aaaccgacta cacaaatcag
     3061 cgatttccat gttgccactc gctttaatga tgatttcagc cgcgctgtac tggaggctga
     3121 agttcagatg tgcggcgagt tgcgtgacta cctacgggta acagtttctt tatggcaggg
     3181 tgaaacgcag gtcgccagcg gcaccgcgcc tttcggcggt gaaattatcg atgagcgtgg
     3241 tggttatgcc gatcgcgtca cactacgtct gaacgtcgaa aacccgaaac tgtggagcgc
     3301 cgaaatcccg aatctctatc gtgcggtggt tgaactgcac accgccgacg gcacgctgat
     3361 tgaagcagaa gcctgcgatg tcggtttccg cgaggtgcgg attgaaaatg gtctgctgct
     3421 gctgaacggc aagccgttgc tgattcgagg cgttaaccgt cacgagcatc atcctctgca
     3481 tggtcaggtc atggatgagc agacgatggt gcaggatatc ctgctgatga agcagaacaa
     3541 ctttaacgcc gtgcgctgtt cgcattatcc gaaccatccg ctgtggtaca cgctgtgcga
     3601 ccgctacggc ctgtatgtgg tggatgaagc caatattgaa acccacggca tggtgccaat
     3661 gaatcgtctg accgatgatc cgcgctggct accggcgatg agcgaacgcg taacgcgaat
     3721 ggtgcagcgc gatcgtaatc acccgagtgt gatcatctgg tcgctgggga atgaatcagg
     3781 ccacggcgct aatcacgacg cgctgtatcg ctggatcaaa tctgtcgatc cttcccgccc
     3841 ggtgcagtat gaaggcggcg gagccgacac cacggccacc gatattattt gcccgatgta
     3901 cgcgcgcgtg gatgaagacc agcccttccc ggctgtgccg aaatggtcca tcaaaaaatg
     3961 gctttcgcta cctggagaga cgcgcccgct gatcctttgc gaatacgccc acgcgatggg
     4021 taacagtctt ggcggtttcg ctaaatactg gcaggcgttt cgtcagtatc cccgtttaca
     4081 gggcggcttc gtctgggact gggtggatca gtcgctgatt aaatatgatg aaaacggcaa
     4141 cccgtggtcg gcttacggcg gtgattttgg cgatacgccg aacgatcgcc agttctgtat
     4201 gaacggtctg gtctttgcgc gatcgcacct cagcgtcgtg actgggaaaa ccctggcgac
     4261 tagtcttgga ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt
     4321 cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccagacg tgtaggctgg
     4381 agctgcttcc ctgcaggagt caggttttat attgcgaaaa ataattggcg aacgaggtaa
     4441 ctggatacct catccgccaa ttaaaatttg ctgtgcaaca ggtgacatca gaggtcatcg
     4501 ttcaaaatgg tatgcgtttt gacacatcca ctatatatcc gtgtcgttct gtccactcct
     4561 gaatcccatt ccagaaattc tctagcgatt ccagaagttt ctcagagtcg gaaagttgac
     4621 cagacattac gaactggcac agatggtcat aacctgaagg aagatctgat tgcttaactg
     4681 cttcagttaa gaccgaagcg ctcgtcgtat aacagatgcg atgatgcaga ccaatcaaca
     4741 tggcacctgc cattgctacc tgcacagtca aggatggtag aaatgttgtc ggtccttgca
     4801 cacgaatatt acgccatttg cctgcatatt caaacagctc ttctacgata agggcacaaa
     4861 tcgcatcgtg gaacgtttgg gcttctaccg atttagcagt ttgatacact ttctctaagt
     4921 atccacctga atcataaatc ggcaaaatag agaaaaattg accatgtgta agcggccaat
     4981 ctgattccac ctgagatgca taatctagta gaatctcttc gctatcaaaa ttcacttcca
     5041 ccttccactc accggttgtc cattcatggc tgaactctgc ttcctctgtt gacatgacac
     5101 acatcatctc aatatccgaa tagggcccat cagtctgacg accaagagag ccataaacac
     5161 caatagcctt aacatcatcc ccatatttat ccaatattcg ttccttaatt tcatgaacaa
     5221 tcttcattct ttcttctcta gtcattatta ttggtccatt cactattctc attccccttt
     5281 cagataattt tagatttgct tttctaaata agaatatttg gagagcaccg ttcttattca
     5341 gctattaaac ccattatatc gggtttttga ggggatttca actgcagcgt tgtccttttc
     5401 cgctgtggtt gtttttgttg gaagtgtgta tcaggtatct gcatagttat tccgaacttc
     5461 caattaatgc aagctggtac cctaaggagg atattcatat gcaagcactt ctgcagtgca
     5521 ggctagcttt tttgtgcaaa agcgtggtaa tttatggtct tttcgagcgg atgaatgaga
     5581 acaaaatcga accacatact acatatataa ccaccgaaag atggtgatca atgatggaac
     5641 gacgaatatt tatccggctt cgccaccgag tgctggcaca tccaggggat attattaccg
     5701 ttggagatgc cgcgcaaata gaagggcagc ttcagctgaa aaagaaactt tcggctatgc
     5761 cgctttatca ggtgagcgaa aaagataaaa atatcgtaat tctggatatc atacaagtcc
     5821 tcagagccat tcatttacaa gacccgacaa ttgatgttca aaccgtaggc ggagcagaaa
     5881 ccattgttga aattcagtat cgaaagcgaa atttatcaac ggttctattt atcggtgtct
     5941 ggctgcttct gtttattgga tcgtgtcttg ccatcatgaa ctttcatgag gatgtaagca
     6001 tgagagatgt tcatatcgca ctatatgaaa tcataaccgg agagaggaat gactatccat
     6061 atttgcttca aatcccatac agcatcggtt tgggactggg gatgatcgtg ttttttaacc
     6121 acatatttaa aaagcgccta aatgaagagc ccagcccgct ggaggttgag atgtttaact
     6181 atcagcttga tctcgatcaa tatgtggcca tgcatgagaa tcaagaaacc ataaaggatc
     6241 tgcatgatcg ttagtgtatt gttcatcatt tttgtcgggc tcggcggagg cataacggtg
     6301 ggggcgggtt tcgttgcttt tttaaccgta atgggaatca ttccgcggct gatgcagctc
     6361 accaaaacaa tgagatttgt tcaggcttat gaagcgacct gactgggtcc ccaataatta
     6421 cgatttaaat tagtagcccg cctaatgagc gggctttttt ttaattcccc tatttgttta
     6481 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     6541 caataatatt gaaaaaggaa gagtatgagc attcagcatt ttcgtgtggc gctgattccg
     6601 ttttttgcgg cgttttgcct gccggtgttt gcgcatccgg aaaccctggt gaaagtgaaa
     6661 gatgcggaag atcaactggg tgcgcgcgtg ggctatattg aactggatct gaacagcggc
     6721 aaaattctgg aatcttttcg tccggaagaa cgttttccga tgatgagcac ctttaaagtg
     6781 ctgctgtgcg gtgcggttct gagccgtgtg gatgcgggcc aggaacaact gggccgtcgt
     6841 attcattata gccagaacga tctggtggaa tatagcccgg tgaccgaaaa acatctgacc
     6901 gatggcatga ccgtgcgtga actgtgcagc gcggcgatta ccatgagcga taacaccgcg
     6961 gcgaacctgc tgctgacgac cattggcggt ccgaaagaac tgaccgcgtt tctgcataac
     7021 atgggcgatc atgtgacccg tctggatcgt tgggaaccgg aactgaacga agcgattccg
     7081 aacgatgaac gtgataccac catgccggca gcaatggcga ccaccctgcg taaactgctg
     7141 acgggtgagc tgctgaccct ggcaagccgc cagcaactga ttgattggat ggaagcggat
     7201 aaagtggcgg gtccgctgct gcgtagcgcg ctgccggctg gctggtttat tgcggataaa
     7261 agcggtgcgg gcgaacgtgg cagccgtggc attattgcgg cgctgggccc ggatggtaaa
     7321 ccgagccgta ttgtggtgat ttataccacc ggcagccagg cgacgatgga tgaacgtaac
     7381 cgtcagattg cggaaattgg cgcgagcctg attaaacatt ggtaaaccga tacaattaaa
     7441 ggctcctttt ggagcctttt tttttggacg acccttgtcc ttttccgctg cataaccctg
     7501 cttcggggtc attatagcga ttttttcggt atatccatcc tttttcgcac gatatacagg
     7561 attttgccaa agggttcgtg tagactttcc ttggtgtatc caacggcgtc agccgggcag
     7621 gataggtgaa gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca
     7681 cctggcggtg ctcaacggga atcctgctct gcgaggctgg ccgtaggccg gccctaccgg
     7741 cgcggcagcg ttacccgtgt cggcggctcc aacggctcgc catcgtccag aaaacacggc
     7801 tcatcgggca tcggcaggcg ctgctgcccg cgccgttccc attcctccgt ttcggtcaag
     7861 gctggcaggt ctggttccat gcccggaatg ccgggctggc tgggcggctc ctcgccgggg
     7921 ccggtcggta gttgctgctc gcccggatac agggtcggga tgcggcgcag gtcgccatgc
     7981 cccaacagcg attcgtcctg gtcgtcgtga tcaaccacca cggcggcact gaacaccgac
     8041 aggcgcaact ggtcgcgggg ctggccccac gccacgcggt cattgaccac gtaggccgac
     8101 acggtgccgg ggccgttgag cttcacgacg gagatccagc gctcggccac caagtccttg
     8161 actgcgtatt ggaccgtccg caaagaacgt ccgatgagct tggaaagtgt cttctggctg
     8221 accaccacgg cgttctggtg gcccatctgc gccacgaggt gatgcagcag cattgccgcc
     8281 gtgggtttcc tcgcaataag cccggcccac gcctcatgcg ctttgcgttc cgtttgcacc
     8341 cagtgaccgg gcttgttctt ggcttgaatg ccgatttctc tggactgcgt ggccatgctt
     8401 atctccatgc ggtaggggtg ccgcacggtt gcggcaccat gcgcaatcag ctgcaacttt
     8461 tcggcagcgc gacaacaatt atgcgttgcg taaaagtggc agtcaattac agattttctt
     8521 taacctacgc aatgagctat tgcggggggt gccgcaatga gctgttgcgt accccccttt
     8581 tttaagttgt tgatttttaa gtctttcgca tttcgcccta tatctagttc tttggtgccc
     8641 aaagaagggc acccctgcgg ggttccccca cgccttcggc gcggctcccc ctccggcaaa
     8701 aagtggcccc tccggggctt gttgatcgac tgcgcggcct tcggccttgc ccaaggtggc
     8761 gctgccccct tggaaccccc gcactcgccg ccgtgaggct cggggggcag gcgggcgggc
     8821 ttcgcccttc gactgccccc actcgcatag gcttgggtcg ttccaggcgc gtcaaggcca
     8881 agccgctgcg cggtcgctgc gcgagccttg acccgccttc cacttggtgt ccaaccggca
     8941 agcgaagcgc gcaggccgca ggccggaggc ttttccccag agaaaattaa aaaaattgat
     9001 ggggcaaggc cgcaggccgc gcagttggag ccggtgggta tgtggtcgaa ggctgggtag
     9061 ccggtgggca atccctgtgg tcaagctcgt gggcaggcgc agcctgtcca tcagcttgtc
     9121 cagcagggtt gtccacgggc cgagcgaagc gagccagccg gtggccgctc gcggccatcg
     9181 tccacatatc cacgggctgg caagggagcg cagcgaccgc gcagggcgaa gcccggagag
     9241 caagcccgta gggggaggcg atctcgagag cttgcatgct gaacttggag gacctttcct
     9301 ttatggacag ctcggggctt ggcgttattt taggaagata caagcaaatt aagcaaattg
     9361 gcggagaaat ggttgtttgc gctatctctc ctgcggtgaa gcgattgttt gatatgtcgg
     9421 gtctgtttaa aattatccga tttgaacaat ctgaacagca ggcactcctg acactggggg
     9481 tggcatcatg aaaaatgaaa tgcaccttga gttttctgcc ctcagtcaga atgaatcgtt
     9541 cgcccgtgtg acagttgctt catttatagc tcagctggac ccgacaatgg atgaactgac
     9601 tgaaatcaaa acagtcgtgt cagaggctgt cacgaatgcg attatccatg gatatgaaga
     9661 gaactgtgaa gggaaagttt acatttcagt gacgctggaa gatcatgtcg tatatatgac
     9721 tattcgtgat gaaggcttag gcattacaga tcttgaagaa gcccgtcagc ctctatttac
     9781 gactaagcct gagcttgagc gctctggaat gggctttacc attatggaaa atttcatgga
     9841 tgatgtcagt atcgattcat cgcctgaaat gggaacaacg attcgcttaa caaagcactt
     9901 atcaaaaagc aaagcgcttt gtaattaagg agatttgtta tggattaacc agcttgtaca
     9961 g
//
