LOCUS       sequence-145622-        8452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mKate2 (codon adapted for Streptococcus pneumoniae); under
            control of CSP inducible promoter PssbB. 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 8452)
  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 8452)
  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-PssbB-mKate2
FEATURES             Location/Qualifiers
     source          1..8452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2417..2439)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2525..2665
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2580..2599
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2680..2697)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2851..3439)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2931..2950)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3610..4401,4402..4470))
                     /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     4233..4252
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4471..4575)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4643..4661
                     /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 ccgcctagaa
      241 ttccaatccc gggtaaagct tccagcggcc gcctcaggat attgcagaca cagtagtcta
      301 tgtgaccagt cagcctcgcc gtgttcagat tacagatatg accattatgg ccaatcaaca
      361 ggcgacaggt ttcatgattc ataaaaaata agaaatttcc tcgaaaagtt acaaatttct
      421 gtaacttttt ttgatttcct acgaatagat aagtaggagg aagaaaatat gtataataaa
      481 gttatcatga ttgggcgttt aacgtctaca ccggatccat tgatatcata ttctagagga
      541 ggaaaattaa tgtcagaact tatcaaggaa aatatgcaca tgaaattgta catggaagga
      601 acagtaaata atcaccactt taaatgtacc tcagaaggag aaggaaaacc atatgaaggt
      661 actcaaacca tgcgtattaa ggccgttgaa ggtggaccat tgccttttgc ctttgatatt
      721 cttgctacat cttttatgta cggatcaaag acttttatca atcataccca aggtatccca
      781 gatttcttta aacagtcatt tcctgaagga tttacatggg aacgtgtcac aacttatgaa
      841 gatggtggag tattgacagc aactcaagat acatctttgc aagatggttg tcttatctac
      901 aatgtaaaga tccgtggagt taattttcca agtaatggtc ctgttatgca gaaaaagacc
      961 cttggatggg aagcatctac cgaaacatta tatcctgccg atggtggatt ggaaggtcgt
     1021 gctgatatgg cattgaaact tgtcggtgga ggtcacctta tctgtaattt gaagaccaca
     1081 taccgttcta aaaagccagc taagaatctt aagatgcctg gtgtttacta cgtcgatcgt
     1141 cgtttagaac gtatcaaaga agcagataag gaaacttatg ttgaacagca cgaagtagcc
     1201 gtcgcacgtt attgtgattt gcctagtaaa ttgggacacc gttaataata aggatcctaa
     1261 ctcgagaaag aattcatgag atctcgcact agtcaaggtc ggcaattctg cagtactagg
     1321 acgccgccaa gccagcttaa acccagctca atgagctggg ttttttgttt gttaaaaatg
     1381 aagaagaaac tgtgaagcgt atttatagca aagcactcaa aagtttacct tatgggtgct
     1441 tttttcgtgc ttttttgaaa agacaaaaaa aagaaccttg ccaagcaaga ttcttttgat
     1501 agcgctatcg ctgagcgccg gtcgctacca ttaccagttg gtctggtgtc aaaaataata
     1561 ataaccgggc aggccatgtc tgcccgtatt tcgcgtaagg aaatccatta tgtactattt
     1621 ctggtgatga aatcaacgta acatttaaag ctgtcaaagc caaagtcatg agatggcgta
     1681 tggagcgtaa agctgacaag agcggtgttg cgatgattga gatgaccttc cttgcaccaa
     1741 gtgaattgcc tcaagaaagc actcaatcaa agattcttgt agatggaaaa gaacttgctg
     1801 atttcgctga aaatcgtcaa gactatcaaa ttacctataa aggtcaacgg ccaaaagtct
     1861 cagttgaaga aaacaatcaa gtagcttcaa ctgtggtaga tagtggagaa gatagccttc
     1921 cagtacttgt tcgcctcgtt tcagaaagtg gaaaacaagt caaggaatac cgtatccagt
     1981 tgactaagga aaaaccagtt tctgctgtac aagaagatct tccaaaactc gaatttgttg
     2041 aaaaagattt ggcctacaag acagttgaga aaaaagattc aacactgtat ctaggtgaaa
     2101 ctcgtgtaga acaagaagga aaagttggaa aagaacgtat ctttacagtg attaatcctg
     2161 atggaagtaa ggaagaaaaa ctccgtgaag tggtagaagt tccgacagac cgcatcgtct
     2221 tggttggaac caaaccagta gctcaagaag ctaaaaaacc acaagtgtca gaaaaagcag
     2281 atacaaaacc aattgattca agtgaagctg atcaaactaa taaagcccag ttaccaaata
     2341 caggtagtgc ggcaagccaa gcagcagtag cagcaggttt agcctgcctc gcgcgtttcg
     2401 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     2461 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     2521 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc
     2581 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     2641 cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2701 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2761 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2821 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2881 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2941 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3001 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3061 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3121 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3181 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3241 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     3301 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3361 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3421 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3481 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3541 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3601 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3661 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3721 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3781 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3841 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3901 tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat
     3961 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4021 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4081 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4141 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4201 accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt
     4261 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4321 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4381 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4441 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4501 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4561 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     4621 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt
     4681 aattccttct taacgcccca agttcatcac caatgacatc aactcacatg aactacatga
     4741 tgaacccagt tatcatggtt ttggataaga tttttgaaaa attcttccca ggccttgata
     4801 aatatgactt tgatgctgct aaattgaaca agaaaatcgg tttctgggga tctaaattct
     4861 tcatcggttt catccttggt atcgttatcg gtattatggg aactccacat ccaattgcag
     4921 gtgttgcaga tgcagataaa tggcgtcttg ttatcaaagg atggttgtct cttggtttga
     4981 ctgccggtgt atctttggaa ctcttctcac ttatcggttc atggttcatc gcagccgtag
     5041 aaccactatc acaaggtatt acaaacgttg ctactaaacg tcttcaagga cgtaaattca
     5101 atatcggtct tgactggcca ttcatcgctg gtcgtgctga aatctgggct tgtgccaacg
     5161 tacttgcacc aatcatgttg attgaagcag tgcttctttc aaaagttgga aatggtatct
     5221 tgccacttgc aggtatcatc gctatgggtg ttactccagc tctcttggtt gtaactcgtg
     5281 gtaaattgct ccgtatgatt atcttcggaa cactcttgtt gccactcttc cttctttcag
     5341 gtacacttat tgcaccattt gcaacagaac ttgctaaagg tgtaggtgcc ttcccagaag
     5401 gtgtgagcca aactcaattg attactcact ctactcttga aggaccaatc gaaaaacttc
     5461 ttggttggac aattggtaac actacaactg gtgatatcaa agcaatcctt ggtgcagtag
     5521 ccttccttgt attctatatc ggtatctttg cttggtacag aaaacaaatg atcaaacgta
     5581 acgaagagta cgcagcaaaa gcaaaataat gcgctccgct agctttacag acaaagaact
     5641 atccttaatg gaagcactag caaaaggtat caatgaagca agaaatattg aaggctagtc
     5701 agtaaaattc agataaacaa aaagagccga taagatgaga gtttcatttc tcaaattatc
     5761 ggctctgcgt cttggcgtct ggctctttga ttgttattat attcattttt tgaacattaa
     5821 ttaaagtttc gttcttacat ttggggcagt ataaagggaa atttttaagt atagtatcca
     5881 ctcgtattcg tagtcttgtt ttatttccac aaataggaca caatatccac ttgtagttta
     5941 taataacaat ctcctccttt ccactttaat tcaaatctat attaaagaat atttcatctt
     6001 atttaataag aaaccatatt tatataacaa cataaaacgc actaagttat tttattgaac
     6061 atatatctta ctttatctat ctgactattt agacgacggg tctggcaaac aggttcgcca
     6121 gtggtaacct gatatccttt tagctctgct aaacaaacac taagcccatt tgtaaaaaaa
     6181 gttaaatcat tgcgataatc ttgaatacat cgagcaggaa tttctccaat aataatgacc
     6241 tcattatttt tcagttgagt atttacgata tttgcacaat atttgggagc atcgttatat
     6301 gcccgtgaaa gatattcctg tggtgcataa actttaaaac taagatatgg ctctaacaat
     6361 tctgttccag cttttctaaa ggcttgctcc aatacaatag gagcaagcat ccgaaaatct
     6421 gctggggtac taacagggct atagtataag ccatacttaa aacagatttt acagtccgtc
     6481 acattccaac catacaatcc ttgttcacaa ccatagcgta tcccttccat aactgcattt
     6541 tgaaatgatt gatttaagta tccaagagaa accgagctct catactgcat tccacttccc
     6601 aacggaagcg gtgatacaga taaaccaatg gaagcccaga aaggatttgg cggcacttcg
     6661 atgtgaatgg tatattctgc attttttaac ggtctctcca tataaatgac tgtaggctct
     6721 tttagttcta tctccacatg atacttttct tgcaacagtg cactaatcac ttccatttgt
     6781 actttcccta agaaagaaag tataatttca tgtgtcgtag aatccacgta atatcgtaga
     6841 agcggatcac tatctgagat ttccaaaagg gcatcaagca acatttctct ctgttcaggt
     6901 ttactcggtt caacagttgt ttgtagtaga gggtgcggat tttcaatctt ttttctctgt
     6961 ggcaatagtt ttgtatctcc aagaacacta tttaacttca aaaactcatt ttgcaaaata
     7021 acaatttctc cagaataagc tctatcaatc ttacataatt caccatttat tgaagtatac
     7081 atttctgtaa cttttatttt ttctttttct gatactctaa ccgaatctcg taaatgtagt
     7141 actccactat aaaggcgtat atatgcaaga cgttgtcttt tttttgtata ttcaattttg
     7201 aaaacatttc cgcaaagttc agacggacct cgatgtgttg atgaataaaa tttattagta
     7261 ataacttcta taaggttatc aatccctata ttactttttg cacttccatg ataaagaggg
     7321 aacagagaac aattctgaaa tcttatgctt tcctcttgtt cgagttccaa tgcttctaat
     7381 gatttaccgg acatatattt ctctaaaagg tcatcgtttc cctctattac cgtatcccat
     7441 tgttcagatt cggtaaagtt cgtcacacac acattaggat acagttctac cttctgtttg
     7501 attacaattt cggcagaaag tttctcttta atatcctgat aaaccgttga taaatcaatt
     7561 ccattttggt caatcttatt gataaaaaag attgtgggaa tccccatttt cctaagtgca
     7621 tgaaataata tacgagtttg tgcttgtacg ccatcttttg cagaaatcag tagaattgcc
     7681 ccatctaaaa ctgataatga acgatatact tctgctaaga aatccatatg tcctggcgtg
     7741 tctatgatgt tcaccttcgt attttcccac tgaaaagagg ttattcctgt ctgaattgta
     7801 attcctctct gacgttctaa aagcgtatta tccgtcctcg ttgtaccttt gtccacgctt
     7861 cctaattctg taatcgctcc actgttatat aataagcttt ctgttaaggt agtttttcct
     7921 gcatcaacat gagctaaaac tccaatatta ataattttca tgtgattttc ctccattcaa
     7981 aagcccaaaa gggcataaaa atcccagtga taaatactct tatcactggg atttttatgc
     8041 ataaccatag gcatacaaag catacagata ttctccggat actttagaat cacatgataa
     8101 aggtattctt aaactgggta caaaaaacta agccctccta aaaaaggaca tccaattatt
     8161 tgttcccgct atcaaattga cagtttattt aagaatacct tgccgcatat ttattaactc
     8221 cttttaaata gatacttaaa ttatagcacg taagagcata tttgtaaagg aatctccaat
     8281 tttttatcaa aaagagtaca tgattacaaa gtatctgtaa tcatgtacca atatttgtta
     8341 ttttacaatc ttccaattat cttgtttttc taatatcaac tcaaattgcg agatttgggt
     8401 tgcctttgtt tcttgatcta aatatttcac agcgacattc acgattactt gg
//
