LOCUS       sequence-85535-e        7327 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Empty vector, integration at lacA, ampr, mlsr.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7327)
  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 7327)
  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: pBS2E
FEATURES             Location/Qualifiers
     source          1..7327
                     /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"
     promoter        1733..1837
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1838..1906,1907..2698)
                     /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     complement(2056..2075)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2869..3457
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3358..3377
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3611..3628
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3643..3783
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3709..3728)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3869..3891
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3885..4076)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      complement(5712..5739)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5758..5844)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        7098..7202
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(7248..7282)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
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 agagatatcg atttcaagct atatttggag
     1141 ttgagcctct tgaaacggac accctgtatc cgaaggatcg aaacgctgtc agctaccgca
     1201 gccaaatata tgaaatgaag gattatgcaa ccgtgattga tgtaaagaca gcttcagtgg
     1261 aagcggtgta tcaagaagat ttttatgcgc gcacgccagc ggtcacaagc catgagtatc
     1321 agcagggcaa ggcgtatttt atcggcgcgc gtttggagga tcaatttcag cgtgatttct
     1381 atgagggtct gatcacagac ctgtctctct ctccagtttt tccggttcgg cacggaaaag
     1441 gcgtctccgt acaagcgagg caggatcagg acaatgatta tatttttgtc atgaatttca
     1501 cggaagaaaa acagctggtc acgtttgatc agagtgtgaa ggacataatg acaggagaca
     1561 tattgtcagg cgacctgacg atggaaaagt atgaagtgag aattgtcgta aacacacatt
     1621 agcccatcat tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     1681 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     1741 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     1801 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     1861 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     1921 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     1981 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     2041 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     2101 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     2161 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     2221 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     2281 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     2341 tgaagccata ccaaacgacg agcgtgacac cacgatgccg gcagcaatgg caacaacgtt
     2401 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     2461 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     2521 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     2581 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     2641 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     2701 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     2761 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     2821 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2881 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2941 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3001 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3061 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3121 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3181 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3241 gagataccta cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3301 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3361 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3421 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3481 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3541 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3601 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     3661 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     3721 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     3781 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     3841 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     3901 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat
     3961 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat
     4021 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcacttga
     4081 tgcctccgtg taagggggaa tttctgttca tgggggtaat gataccgatg aaacgagaga
     4141 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     4201 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     4261 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     4321 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     4381 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     4441 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
     4501 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc
     4561 tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat atgttctgcc
     4621 aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca attcttggag
     4681 tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca
     4741 tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta caatccatgc
     4801 caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca gcggtccagt
     4861 gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct gatggtcgtc
     4921 atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag
     4981 aagaatcata atggggaagg ccatccagcc tcgcgtcgcg actaagaaaa tgccgtcaaa
     5041 tccgctcgcc atgacttcac taacgatgcc tttgaaaatc ttcaagttct tttctactaa
     5101 ttcaaggcgt gtctcaccag gtttttggtt tgctccggcg caaatgcaga caatatcagg
     5161 atggggtgat gtcaaagctt gaaaaaacgc acgtaacaaa agcaaaattt atgctccatg
     5221 ggggagacta caaccccgat cagtggctgg atcggcccga tattttagct gacgatatca
     5281 aactgatgaa gctttctcat acgaatacgt tttctgtcgg catttttgca tggagcgcac
     5341 ttgagccgga ggagggcgta tatcaatttg aatggctgga tgatattttt gagcggattc
     5401 acagtatagg cggccgggtc atattagcaa cgccgagcgg agcccgtccg gcctggctgt
     5461 cgcaaaccta tccggaagtt ttgcgcgtca atgcctcccg cgtcaaacag ctgcacggcg
     5521 gaaggcacaa ccactgcctc acatctaaag tctaccgaga aaaaacacgg cacatcaacc
     5581 gcttattagc agaacgatac ggacatcacc cggcgctgtt aatgtggcac atttcaaacg
     5641 aatacggggg agattgccac tgtgaatcga tgcggccgct ctaggagtta acaagagttt
     5701 gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagctaga gcggcggatt
     5761 tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttt
     5821 cgactgagcc tttcgtttta tttgatgcct caagctagag agtcctagag tctagggacc
     5881 tctttagctc cttggaagct gtcagtagta tacctaataa tttatctaca ttccctttag
     5941 taacgtgtaa ctttccaaat ttacaaaagc gactcataga attatttcct cccgttaaat
     6001 aatagataac tattaaaaat agacaatact tgctcataag taacggtact taaattgttt
     6061 actttggcgt gtttcattgc ttgatgaaac tgatttttag taaacagttg acgatattct
     6121 cgattgaccc attttgaaac aaagtacgta tatagcttcc aatatttatc tggaacatct
     6181 gtggtatggc gggtaagttt tattaagaca ctgtttactt ttggtttagg atgaaagcat
     6241 tccgctggca gcttaagcaa ttgctgaatc gagacttgag tgtgcaagag caaccctagt
     6301 gttcggtgaa tatccaaggt acgcttgtag aatccttctt caacaatcag atagatgtca
     6361 gacgcatggc tttcaaaaac cactttttta ataatttgtg tgcttaaatg gtaaggaata
     6421 ctcccaacaa ttttatacct ctgtttgtta gggaattgaa actgtagaat atcttggtga
     6481 attaaagtga cacgagtatt cagttttaat ttttctgacg ataagttgaa tagatgactg
     6541 tctaattcaa tagacgttac ctgtttactt attttagcca gtttcgtcgt taaatgccct
     6601 ttacctgttc caatttcgta aacggtatcg gtttctttta aattcaattg ttttattatt
     6661 tggttgagta ctttttcact cgttaaaaag ttttgagaat attttatatt tttgttcatg
     6721 taatcactcc ttcttaatta caaattttta gcatctaatt taacttcaat tcctattata
     6781 caaaatttta agatactgca ctatcaacac actcttaagt ttgcttctaa gtcttatttc
     6841 cataacttct tttacgtttc cgccattctt tgctgtttcg atttttatga tatggtgcaa
     6901 gtcagcacga acacgaaccg tcttatctcc cattatatct ttttttgcac tgattggtgt
     6961 atcatttcgt ttttcttttg tgctagagga tcaattcttg aagacgaaag ggcctcgtga
     7021 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     7081 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     7141 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     7201 gtgcggccgc ccgcgggagc tccccgggac gttcttgcca ttgctgcata aaaaacgccc
     7261 ggcggcaacc gagcgttctg aattaattaa tcatcgcgac tgcatctttc gctaaggatg
     7321 atttctg
//
