LOCUS       sequence-85536-e        5642 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Empty vector, integration at thrC, ampr, specr.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5642)
  AUTHORS   Radeck J, Kraft K, Bartels J, Cikovic T, Durr F, Emenegger J, 
            Kelterborn S, Sauer C, Fritz G, Gebhard S, Mascher T
  TITLE     The Bacillus BioBrick Box: generation and evaluation of essential 
            genetic building blocks for standardized work with Bacillus 
            subtilis.
  JOURNAL   J Biol Eng. 2013 Dec 2;7(1):29. doi: 10.1186/1754-1611-7-29.
  PUBMED    24295448
REFERENCE   2  (bases 1 to 5642)
  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: "J Biol 
            Eng."; date: "2013-12-2"; volume: "7(1):29. doi"; pages: " 
            10.1186/1754-1611-7-29"
COMMENT     Sequence Label: pBS4S
FEATURES             Location/Qualifiers
     source          1..5642
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    1..22
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    123..144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(159..164,165..182,183..189)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    197..213
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             231..242
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             249..926
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      971..1042
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1058..1085
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1092..1112
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     rep_origin      complement(3048..3636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3128..3147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3807..4598,4599..4667))
                     /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"
     primer_bind     4430..4449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4668..4772)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4840..4858
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gaattcgcgg ccgcttctag agcaatacgc aaaccgcctc tccccgcgcg ttggccgatt
       61 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
      121 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
      181 cgtatgttgt gtggaattgt gagcggataa caatttcaca catactagag aaagaggaga
      241 aatactagat ggcttcctcc gaagacgtta tcaaagagtt catgcgtttc aaagttcgta
      301 tggaaggttc cgttaacggt cacgagttcg aaatcgaagg tgaaggtgaa ggtcgtccgt
      361 acgaaggtac ccagaccgct aaactgaaag ttaccaaagg tggtccgctg ccgttcgctt
      421 gggacatcct gtccccgcag ttccagtacg gttccaaagc ttacgttaaa cacccggctg
      481 acatcccgga ctacctgaaa ctgtccttcc cggaaggttt caaatgggaa cgtgttatga
      541 acttcgaaga cggtggtgtt gttaccgtta cccaggactc ctccctgcaa gacggtgagt
      601 tcatctacaa agttaaactg cgtggtacca acttcccgtc cgatggtccg gttatgcaga
      661 aaaaaaccat gggttgggaa gcttccaccg aacgtatgta cccggaagac ggtgctctga
      721 aaggtgaaat caaaatgcgt ctgaaactga aagacggtgg tcactacgac gctgaagtta
      781 aaaccaccta catggctaaa aaaccggttc agctgccggg tgcttacaaa accgacatca
      841 aactggacat cacctcccac aacgaagact acaccatcgt tgaacagtac gaacgtgctg
      901 aaggtcgtca ctccaccggt gcttaataac gctgatagtg ctagtgtaga tcgctactag
      961 agccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     1021 ttgtttgtcg gtgaacgctc tctactagag tcacactggc tcaccttcgg gtgggccttt
     1081 ctgcgtttat atactagtag cggccgctgc aggataaaaa atttagaagc caatgaaatc
     1141 tataaataaa ctaaattaag tttatttaat taacaactat ggatataaaa taggtactaa
     1201 tcaaaatagt gaggaggata tatttgaata catacgaaca aattaataaa gtgaaaaaaa
     1261 tacttcggaa acatttaaaa aataacctta ttggtactta catgtttgga tcaggagttg
     1321 agagtggact aaaaccaaat agtgatcttg actttttagt cgtcgtatct gaaccattga
     1381 cagatcaaag taaagaaata cttatacaaa aaattagacc tatttcaaaa aaaataggag
     1441 ataaaagcaa cttacgatat attgaattaa caattattat tcagcaagaa atggtaccgt
     1501 ggaatcatcc tcccaaacaa gaatttattt atggagaatg gttacaagag ctttatgaac
     1561 aaggatacat tcctcagaag gaattaaatt cagatttaac cataatgctt taccaagcaa
     1621 aacgaaaaaa taaaagaata tacggaaatt atgacttaga ggaattacta cctgatattc
     1681 cattttctga tgtgagaaga gccattatgg attcgtcaga ggaattaata gataattatc
     1741 aggatgatga aaccaactct atattaactt tatgccgtat gattttaact atggacacgg
     1801 gtaaaatcat accaaaagat attgcgggaa atgcagtggc tgaatcttct ccattagaac
     1861 atagggagag aattttgtta gcagttcgta gttatcttgg agagaatatt gaatggacta
     1921 atgaaaatgt aaatttaact ataaactatt taaataacag attaaaaaaa ttataaaaaa
     1981 attgaaaaaa tggtggaaac acttttttca atttttttgt tttattattt aatatttggg
     2041 aaatattcat tctaattggt aatcagattt tagaaaacaa taaacccttg cataggggga
     2101 tctcgaggcc ttccgaaaat gcgcggcttt gaagctgagg gtgcggcggc aatcgtgcgc
     2161 aatgaagtga ttgaaaatcc ggaaacaata gcgacagcca ttcgtatcgg aaacccggca
     2221 agctgggaca aagctgtaaa ggcagccgag gaatccaatg ggaaaattga cgaagtcact
     2281 gatgatgaaa tccttcacgc atatcagctg atcgcccgtg tagaaggcgt gtttgcagaa
     2341 ccaggttctt gcgcgtctat cgcaggagtg ctgaaacagg tgaaatccgg agaaattccg
     2401 aaaggcagca aggtcgtagc tgtgttaaca ggaaacggac tgaaagatcc gaacacagcg
     2461 gtcgacattt cagaaatcaa gcctgtcaca ttgccgactg atgaagacag catccttgaa
     2521 tatgtaaaag gagcggcccg tgtatgaacg aagccgacat gctgttctct gtcactgttc
     2581 ccggaagcac agctaaccta ggccccggct ttgattcagt cggaatggcg ctcagcagat
     2641 atttgaagct gaccgtcttt gaaagcgaca aatggtcttt tgaggctgaa acagaaacag
     2701 tcgccggaat taattcgagc tcctttaact tcctcaccag tagttgtatc ggtaccataa
     2761 gtagaagcag caacccaagt agctttacca gcatccggtt caaccagcat agtaagaatc
     2821 ttactggaca tcggcagttc ttcgaacagt gcgccaacta ccagctcttt ctacagttca
     2881 ttcagggcac cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc
     2941 atcctgtctc ttgatccatg gattacgcgc gaccttcagc atcgccggca tgtccccctg
     3001 gcggacggga agtatccagc tcgaggtcgg gccgcgttgc tggcgttttt ccataggctc
     3061 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3121 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3181 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3241 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3301 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3361 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3421 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3481 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3541 gttgatagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3601 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3661 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3721 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3781 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3841 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3901 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3961 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4021 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4081 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgccggcat cgtggtgtca
     4141 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4201 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4261 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4321 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4381 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg
     4441 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4501 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4561 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4621 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4681 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4741 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     4801 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     4861 tttcgtcttc aagaattaat tcatgtaaaa gatgaggttg gttcattctc gaaaattaca
     4921 tctgtgttct cagagcgggg cgtgacgttt gaaaaaatcc ttcagctgcc aattaaaggc
     4981 catgatgagt tagctgaaat cgtaattgtc acacatcata catcagaagc tgatttcagt
     5041 gatatcctgc aaaacctaaa tgatttggaa gtcgttcaag aagtcaaaag cacatatcgt
     5101 gtagaaggga acggttggag ctaatgtgga aaggacttat ccatcaatat aaagaatttt
     5161 tacctgtaac agatcaaaca ccggcgctaa ctttacatga aggaaacaca cctcttattc
     5221 acctgccgaa gctgtctgag cagctcggaa ttgagcttca tgtcaaaacg gaaggcgtca
     5281 atcctacggg atcatttaaa gatcgcggaa tggttatggc tgtggcaaag gcaaaagaag
     5341 aaggcaatga cacgattatg tgcgcgtcaa caggtaacac ttccgctgct gcggcagcat
     5401 atgcagcccg tgctaacatg aaatgcattg tcatcatccc gaacggaaaa attgcatttg
     5461 gaaaactcgc tcaagctgtc atgtacggag ccgagattat cgcaattgac ggaaactttg
     5521 acgatgcgct taaaattgtc cgttccatct gtgagaaatc accgattgcc cttgtcaact
     5581 cagtcaaccc ttaccgcatt gaaggccaaa aaactgctgc cttcggatcc tagaagctta
     5641 tc
//
