LOCUS       sequence-85534-e        7171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty vector, integration at amyE, ampr, cmr.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7171)
  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 7171)
  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: pBS1C
FEATURES             Location/Qualifiers
     source          1..7171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1149..1168)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(3583..4171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3663..3682)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4342..5133,5134..5202))
                     /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     4965..4984
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5203..5307)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5375..5393
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    6065..6086
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    6187..6208
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6223..6228,6229..6246,6247..6253)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6261..6277
                     /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             6295..6306
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             6313..6990
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      7035..7106
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7122..7149
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    join(7156..7171,1..5)
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
ORIGIN
        1 tgcagtccgg caaaaaaggg caaggtgtca attctcatgt ttgacagctt atcatcggca
       61 atagttaccc ttattatcaa gataagaaag aaaaggattt ttcgctacgc tcaaatcctt
      121 taaaaaaaca caaaagacca cattttttaa tgtggtcttt attcttcaac taaagcaccc
      181 attagttcaa caaacgaaaa ttggataaag tgggatattt ttaaaatata tatttatgtt
      241 acagtaatat tgacttttaa aaaaggattg attctaatga agaaagcaga caagtaagcc
      301 tcctaaattc actttagata aaaatttagg aggcatatca aatgaacttt aataaaattg
      361 atttagacaa ttggaagaga aaagagatat ttaatcatta tttgaaccaa caaacgactt
      421 ttagtataac cacagaaatt gatattagtg ttttataccg aaacataaaa caagaaggat
      481 ataaatttta ccctgcattt attttcttag tgacaagggt gataaactca aatacagctt
      541 ttagaactgg ttacaatagc gacggagagt taggttattg ggataagtta gagccacttt
      601 atacaatttt tgatggtgta tctaaaacat tctctggtat ttggactcct gtaaagaatg
      661 acttcaaaga gttttatgat ttataccttt ctgatgtaga gaaatataat ggttcgggga
      721 aattgtttcc caaaacacct atacctgaaa atgctttttc tctttctatt attccatgga
      781 cttcatttac tgggtttaac ttaaatatca ataataatag taattacctt ctacccatta
      841 ttacagcagg aaaattcatt aataaaggta attcaatata tttaccgcta tctttacagg
      901 tacatcattc tgtttgtgat ggttatcatg caggattgtt tatgaactct attcaggaat
      961 tgtcagatag gcctaatgac tggcttttat aatatgagat aatgccgact gtacttttta
     1021 cagtcggttt tctaatgtca ctaacctgcc ccgttagttg aagaaggttt ttatattaca
     1081 gctccagatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
     1141 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct
     1201 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg
     1261 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
     1321 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgacatg gatgagcgat
     1381 gatgatatcc gtttaggctg ggcggtgata gcttctcgtt caggcagtac gcctcttttc
     1441 ttttccagac ctgagggagg cggaaatggt gtgaggttcc cggggaaaag ccaaataggc
     1501 gatcgcggga gtgctttatt tgaagatcag gctatcactg cggtcaatag atttcacaat
     1561 gtgatggctg gacagcctga ggaactctcg aacccgaatg gaaacaacca gatatttatg
     1621 aatcagcgcg gctcacatgg cgttgtgctg gcaaatgcag gttcatcctc tgtctctatc
     1681 aatacggcaa caaaattgcc tgatggcagg tatgacaata aagctggagc gggttcattt
     1741 caagtgaacg atggtaaact gacaggcacg atcaatgcca ggtctgtagc tgtgctttat
     1801 cctgatgata ttgcaaaagc gcctcatgtt ttccttgaga attacaaaac aggtgtaaca
     1861 cattctttca atgatcaact gacgattacc ttgcgtgcag atgcgaatac aacaaaagcc
     1921 gtttatcaaa tcaataatgg accagagacg gcgtttaagg atggagatca attcacaatc
     1981 ggaaaaggag atccatttgg caaaacatac accatcatgt taaaaggaac gaacagtgat
     2041 ggtgtaacga ggaccgagaa atacagtttt gttaaaagag atccagcgtc ggccaaaacc
     2101 atcggctatc aaaatccgaa tcattggagc caggtaaatg cttatatcta taaacatgat
     2161 gggagccgag taattgaatt gaccggatct tggcctggaa aaccaatgac taaaaatgca
     2221 gacggaattt acacgctgac gctgcctgcg gacacggata caaccaacgc aaaagtgatt
     2281 tttaataatg gcagcgccca agtgcccggt cagaatcagc ctggctttga ttacgtgcta
     2341 aatggtttat ataatgactc gggcttaagc ggttctcttc cccattgagg gcaaggctag
     2401 acgggactta ccgaaagaaa ccatcaatga tggtttcttt tttgttcata aatcagacaa
     2461 aacttttctc ttgcaaaagt ttgtgaagtg ttgcacaata taaatgtgaa atacttcaca
     2521 aacaaaaaga catcaaagag aaacataccc tgcaaggatg ctgatattgt ctgcatttgc
     2581 gccggagcaa accaaaaacc tggtgagaca cgccttgaat tagtagaaaa gaacttgaag
     2641 attttcaaag gcatcgttag tgaagtcatg gcgagcggat ttgacggcat tttcttagtc
     2701 gcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg
     2761 atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt
     2821 caaggatcgc tcgcggctct taccagccta acttcgatca ctggaccgct gatcgtcacg
     2881 gcgatttatg ccgcctcggc gagcacatgg aacgggttgg catggattgt aggcgccgcc
     2941 ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc cacctactga
     3001 agtggatttc tttaagagct cctttaactt cctcaccagt agttgtatcg gtaccataag
     3061 tagaagcagc aacccaagta gctttaccag catccggttc aaccagcata gtaagaatct
     3121 tactggacat cggcagttct tcgaacagtg cgccaactac cagctctttc tccagaatgg
     3181 gctatacctc ttttacctaa agagcatgta actttactgg atatagctag aaaaggctat
     3241 cggggagagt gtgatgataa gtgggaagga ctatattcaa aggtgaaagc actcgttaag
     3301 tatatgaaaa attctataga aacttctctc aattaggcta attttattgc aataacaggt
     3361 gcttactttt ctggagttct ttagcaaatt tttttattag ctgaacttag tattagtggc
     3421 catactcctc caatccaaag ctatttagaa agattactat atcctcaaac aggcggtaac
     3481 cggcctcttc atcgggaatg cgcgcgacct tcagcatcgc cggcatgtcc ccctggcgga
     3541 cgggaagtat ccagctcgag gtcgggccgc gttgctggcg tttttccata ggctccgccc
     3601 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3661 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3721 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3781 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3841 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3901 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3961 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4021 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttga
     4081 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4141 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4201 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4261 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4321 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4381 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4441 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4501 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4561 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4621 gccagttaat agtttgcgca acgttgttgc cattgctgcc ggcatcgtgg tgtcacgctc
     4681 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4741 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4801 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4861 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4921 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca
     4981 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5041 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5101 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5161 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5221 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5281 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5341 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5401 tcttcaagaa ttaacaaaat tctccagtct tcacatcggt ttgaaaggag gaagcggaag
     5461 aatgaagtaa gagggatttt tgactccgaa gtaagtcttc aaaaaatcaa ataaggagtg
     5521 tcaagaatgt ttgcaaaacg attcaaaacc tctttactgc cgttattcgc tggattttta
     5581 ttgctgtttc atttggttct ggcaggaccg gcggctgcga gtgctgaaac ggcgaacaaa
     5641 tcgaatgagc ttacagcacc gtcgatcaaa agcggaacca ttcttcatgc atggaattgg
     5701 tcgttcaata cgttaaaaca caatatgaag gatattcatg atgcaggata tacagccatt
     5761 cagacatctc cgattaacca agtaaaggaa gggaatcaag gagataaaag catgtcgaac
     5821 tggtactggc tgtatcagcc gacatcgtat caaattggca accgttactt aggtactgaa
     5881 caagaattta aagaaatgtg tgcagccgct gaagaatatg gcataaaggt cattgttgac
     5941 gcggtcatca atcataccac cagtgattat gccgcgattt ccaatgaggt taagagtatt
     6001 ccaaactgga cacatggaaa cacacaaatt aaaaactggt ctgatcggat cctagaagct
     6061 tatcgaattc gcggccgctt ctagagcaat acgcaaaccg cctctccccg cgcgttggcc
     6121 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     6181 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     6241 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacatact agagaaagag
     6301 gagaaatact agatggcttc ctccgaagac gttatcaaag agttcatgcg tttcaaagtt
     6361 cgtatggaag gttccgttaa cggtcacgag ttcgaaatcg aaggtgaagg tgaaggtcgt
     6421 ccgtacgaag gtacccagac cgctaaactg aaagttacca aaggtggtcc gctgccgttc
     6481 gcttgggaca tcctgtcccc gcagttccag tacggttcca aagcttacgt taaacacccg
     6541 gctgacatcc cggactacct gaaactgtcc ttcccggaag gtttcaaatg ggaacgtgtt
     6601 atgaacttcg aagacggtgg tgttgttacc gttacccagg actcctccct gcaagacggt
     6661 gagttcatct acaaagttaa actgcgtggt accaacttcc cgtccgatgg tccggttatg
     6721 cagaaaaaaa ccatgggttg ggaagcttcc accgaacgta tgtacccgga agacggtgct
     6781 ctgaaaggtg aaatcaaaat gcgtctgaaa ctgaaagacg gtggtcacta cgacgctgaa
     6841 gttaaaacca cctacatggc taaaaaaccg gttcagctgc cgggtgctta caaaaccgac
     6901 atcaaactgg acatcacctc ccacaacgaa gactacacca tcgttgaaca gtacgaacgt
     6961 gctgaaggtc gtcactccac cggtgcttaa taacgctgat agtgctagtg tagatcgcta
     7021 ctagagccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     7081 tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc
     7141 ctttctgcgt ttatatacta gtagcggccg c
//
