LOCUS       sequence-125508-        8333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry (codon adapted for Bacillus subtilis); under 
            control of Zinc inducible promoter PZn. Plasmid integrates in S. 
            pneumoniae bgaA locus by double cross-over..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8333)
  AUTHORS   Beilharz K, van Raaphorst R, Kjos M, Veening JW
  TITLE     Red fluorescent proteins for gene expression and protein 
            localization studies in Streptococcus pneumoniae and efficient 
            transformation with Gibson assembled DNA.
  JOURNAL   Appl Environ Microbiol. 2015 Aug 7. pii: AEM.02033-15.
  PUBMED    26253684
REFERENCE   2  (bases 1 to 8333)
  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: "Appl 
            Environ Microbiol. 2015 Aug 7. pii: AEM.02033-15."
COMMENT     Sequence Label: pKB01_mCherry
FEATURES             Location/Qualifiers
     source          1..8333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2298..2320)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2406..2546
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2461..2480
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2561..2578)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2732..3320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2812..2831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3491..4282,4283..4351))
                     /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     4114..4133
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4352..4456)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4524..4542
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctagcgctat atgcgttgat gcaatttcta tgcgcacccg ttctcggagc actgtccgac
       61 cgctttggcc gccgcccagt cctgctcgct tcgctacttg gagccactat cgactacgcg
      121 atcatggcga ccacacccgt cctgtggatc tatcgatgca tgcagccggt accatcacaa
      181 gcactttggg acgttctccc ttagtgcttt tttgatttct cataggccgg cctgttagtc
      241 atatggacac ttaaggcaaa ttgttcagaa ctgaataaag ctgacgtttt gcttctatcc
      301 tttctttgag ttttagtgga taatgataat gaacaaggtg ttcataaatc tattataaca
      361 agaattccaa tcccgggtaa agcttccagc ggccgcatag gatccattga tatcatattc
      421 tagatactga ttaactaata aggaggacaa acatgagcaa aggagaagaa gataacatgg
      481 caatcatcaa agaatttatg cgtttcaaag ttcacatgga aggttctgta aacggacacg
      541 aatttgaaat tgaaggtgaa ggtgaaggcc gtccttatga aggaacacaa acggcaaagc
      601 tgaaagtaac aaaaggcgga ccgcttccgt ttgcatggga tatcctttct ccgcaattca
      661 tgtacggttc aaaagcatac gtgaagcatc cggctgatat tcctgattat ttgaagctgt
      721 cattccctga aggcttcaaa tgggagcgtg tgatgaactt tgaagatggc ggtgttgtta
      781 ctgttactca agattcaagc cttcaagacg gtgaatttat ttacaaagtg aagctgcgcg
      841 gaacaaactt cccatctgac ggacctgtca tgcaaaagaa aacaatgggc tgggaagcaa
      901 gctctgaacg catgtatcca gaggacggtg ctttaaaagg agaaatcaaa cagcgtttga
      961 agctgaaaga cggcggacac tatgacgctg aagtgaaaac aacttacaaa gcgaaaaagc
     1021 cggttcagct tccaggtgct tacaacgtaa acatcaaact tgatattaca agccacaatg
     1081 aagattatac gattgttgaa caatatgaac gcgctgaagg ccgtcattca actggcggaa
     1141 tggatgagct ttacaaataa taatgagcac tagtcaaggt cggcaattct gcagtactag
     1201 gacgccgcca agccagctta aacccagctc aatgagctgg gttttttgtt tgttaaaaat
     1261 gaagaagaaa ctgtgaagcg tatttatagc aaagcactca aaagtttacc ttatgggtgc
     1321 ttttttcgtg cttttttgaa aagacaaaaa aaagaacctt gccaagcaag attcttttga
     1381 tagcgctatc gctgagcgcc ggtcgctacc attaccagtt ggtctggtgt caaaaataat
     1441 aataaccggg caggccatgt ctgcccgtat ttcgcgtaag gaaatccatt atgtactatt
     1501 tctggtgatg aaatcaacgt aacatttaaa gctgtcaaag ccaaagtcat gagatggcgt
     1561 atggagcgta aagctgacaa gagcggtgtt gcgatgattg agatgacctt ccttgcacca
     1621 agtgaattgc ctcaagaaag cactcaatca aagattcttg tagatggaaa agaacttgct
     1681 gatttcgctg aaaatcgtca agactatcaa attacctata aaggtcaacg gccaaaagtc
     1741 tcagttgaag aaaacaatca agtagcttca actgtggtag atagtggaga agatagcctt
     1801 ccagtacttg ttcgcctcgt ttcagaaagt ggaaaacaag tcaaggaata ccgtatccag
     1861 ttgactaagg aaaaaccagt ttctgctgta caagaagatc ttccaaaact cgaatttgtt
     1921 gaaaaagatt tggcctacaa gacagttgag aaaaaagatt caacactgta tctaggtgaa
     1981 actcgtgtag aacaagaagg aaaagttgga aaagaacgta tctttacagt gattaatcct
     2041 gatggaagta aggaagaaaa actccgtgaa gtggtagaag ttccgacaga ccgcatcgtc
     2101 ttggttggaa ccaaaccagt agctcaagaa gctaaaaaac cacaagtgtc agaaaaagca
     2161 gatacaaaac caattgattc aagtgaagct gatcaaacta ataaagccca gttaccaaat
     2221 acaggtagtg cggcaagcca agcagcagta gcagcaggtt tagcctgcct cgcgcgtttc
     2281 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     2341 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     2401 cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg
     2461 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     2521 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc
     2581 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2641 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2701 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2761 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2821 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2881 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2941 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3001 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3061 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3121 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     3181 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3241 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3301 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3361 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3421 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3481 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3541 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3601 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3661 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3721 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3781 gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta
     3841 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3901 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3961 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4021 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4081 gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt
     4141 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4201 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4261 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4321 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4381 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4441 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     4501 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcaagaat
     4561 taattccttc ttaacgcccc aagttcatca ccaatgacat caactcacat gaactacatg
     4621 atgaacccag ttatcatggt tttggataag atttttgaaa aattcttccc aggccttgat
     4681 aaatatgact ttgatgctgc taaattgaac aagaaaatcg gtttctgggg atctaaattc
     4741 ttcatcggtt tcatccttgg tatcgttatc ggtattatgg gaactccaca tccaattgca
     4801 ggtgttgcag atgcagataa atggcgtctt gttatcaaag gatggttgtc tcttggtttg
     4861 actgccggtg tatctttgga actcttctca cttatcggtt catggttcat cgcagccgta
     4921 gaaccactat cacaaggtat tacaaacgtt gctactaaac gtcttcaagg acgtaaattc
     4981 aatatcggtc ttgactggcc attcatcgct ggtcgtgctg aaatctgggc ttgtgccaac
     5041 gtacttgcac caatcatgtt gattgaagca gtgcttcttt caaaagttgg aaatggtatc
     5101 ttgccacttg caggtatcat cgctatgggt gttactccag ctctcttggt tgtaactcgt
     5161 ggtaaattgc tccgtatgat tatcttcgga acactcttgt tgccactctt ccttctttca
     5221 ggtacactta ttgcaccatt tgcaacagaa cttgctaaag gtgtaggtgc cttcccagaa
     5281 ggtgtgagcc aaactcaatt gattactcac tctactcttg aaggaccaat cgaaaaactt
     5341 cttggttgga caattggtaa cactacaact ggtgatatca aagcaatcct tggtgcagta
     5401 gccttccttg tattctatat cggtatcttt gcttggtaca gaaaacaaat gatcaaacgt
     5461 aacgaagagt acgcagcaaa agcaaaataa tgcgctccgc tagctttaca gacaaagaac
     5521 tatccttaat ggaagcacta gcaaaaggta tcaatgaagc aagaaatatt gaaggctagt
     5581 cagtaaaatt cagataaaca aaaagagccg ataagatgag agtttcattt ctcaaattat
     5641 cggctctgcg tcttggcgtc tggctctttg attgttatta tattcatttt ttgaacatta
     5701 attaaagttt cgttcttaca tttggggcag tataaaggga aatttttaag tatagtatcc
     5761 actcgtattc gtagtcttgt tttatttcca caaataggac acaatatcca cttgtagttt
     5821 ataataacaa tctcctcctt tccactttaa ttcaaatcta tattaaagaa tatttcatct
     5881 tatttaataa gaaaccatat ttatataaca acataaaacg cactaagtta ttttattgaa
     5941 catatatctt actttatcta tctgactatt tagacgacgg gtctggcaaa caggttcgcc
     6001 agtggtaacc tgatatcctt ttagctctgc taaacaaaca ctaagcccat ttgtaaaaaa
     6061 agttaaatca ttgcgataat cttgaataca tcgagcagga atttctccaa taataatgac
     6121 ctcattattt ttcagttgag tatttacgat atttgcacaa tatttgggag catcgttata
     6181 tgcccgtgaa agatattcct gtggtgcata aactttaaaa ctaagatatg gctctaacaa
     6241 ttctgttcca gcttttctaa aggcttgctc caatacaata ggagcaagca tccgaaaatc
     6301 tgctggggta ctaacagggc tatagtataa gccatactta aaacagattt tacagtccgt
     6361 cacattccaa ccatacaatc cttgttcaca accatagcgt atcccttcca taactgcatt
     6421 ttgaaatgat tgatttaagt atccaagaga aaccgagctc tcatactgca ttccacttcc
     6481 caacggaagc ggtgatacag ataaaccaat ggaagcccag aaaggatttg gcggcacttc
     6541 gatgtgaatg gtatattctg cattttttaa cggtctctcc atataaatga ctgtaggctc
     6601 ttttagttct atctccacat gatacttttc ttgcaacagt gcactaatca cttccatttg
     6661 tactttccct aagaaagaaa gtataatttc atgtgtcgta gaatccacgt aatatcgtag
     6721 aagcggatca ctatctgaga tttccaaaag ggcatcaagc aacatttctc tctgttcagg
     6781 tttactcggt tcaacagttg tttgtagtag agggtgcgga ttttcaatct tttttctctg
     6841 tggcaatagt tttgtatctc caagaacact atttaacttc aaaaactcat tttgcaaaat
     6901 aacaatttct ccagaataag ctctatcaat cttacataat tcaccattta ttgaagtata
     6961 catttctgta acttttattt tttctttttc tgatactcta accgaatctc gtaaatgtag
     7021 tactccacta taaaggcgta tatatgcaag acgttgtctt ttttttgtat attcaatttt
     7081 gaaaacattt ccgcaaagtt cagacggacc tcgatgtgtt gatgaataaa atttattagt
     7141 aataacttct ataaggttat caatccctat attacttttt gcacttccat gataaagagg
     7201 gaacagagaa caattctgaa atcttatgct ttcctcttgt tcgagttcca atgcttctaa
     7261 tgatttaccg gacatatatt tctctaaaag gtcatcgttt ccctctatta ccgtatccca
     7321 ttgttcagat tcggtaaagt tcgtcacaca cacattagga tacagttcta ccttctgttt
     7381 gattacaatt tcggcagaaa gtttctcttt aatatcctga taaaccgttg ataaatcaat
     7441 tccattttgg tcaatcttat tgataaaaaa gattgtggga atccccattt tcctaagtgc
     7501 atgaaataat atacgagttt gtgcttgtac gccatctttt gcagaaatca gtagaattgc
     7561 cccatctaaa actgataatg aacgatatac ttctgctaag aaatccatat gtcctggcgt
     7621 gtctatgatg ttcaccttcg tattttccca ctgaaaagag gttattcctg tctgaattgt
     7681 aattcctctc tgacgttcta aaagcgtatt atccgtcctc gttgtacctt tgtccacgct
     7741 tcctaattct gtaatcgctc cactgttata taataagctt tctgttaagg tagtttttcc
     7801 tgcatcaaca tgagctaaaa ctccaatatt aataattttc atgtgatttt cctccattca
     7861 aaagcccaaa agggcataaa aatcccagtg ataaatactc ttatcactgg gatttttatg
     7921 cataaccata ggcatacaaa gcatacagat attctccgga tactttagaa tcacatgata
     7981 aaggtattct taaactgggt acaaaaaact aagccctcct aaaaaaggac atccaattat
     8041 ttgttcccgc tatcaaattg acagtttatt taagaatacc ttgccgcata tttattaact
     8101 ccttttaaat agatacttaa attatagcac gtaagagcat atttgtaaag gaatctccaa
     8161 ttttttatca aaaagagtac atgattacaa agtatctgta atcatgtacc aatatttgtt
     8221 attttacaat cttccaatta tcttgttttt ctaatatcaa ctcaaattgc gagatttggg
     8281 ttgcctttgt ttcttgatct aaatatttca cagcgacatt cacgattact tgg
//
