LOCUS       sequence-125507-        8330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tagRFP (codon adapted for Streptococcus pneumoniae); 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 8330)
  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 8330)
  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_tagRFP
FEATURES             Location/Qualifiers
     source          1..8330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1876..1898)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1984..2124
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2039..2058
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2139..2156)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2310..2898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2390..2409)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3069..3860,3861..3929))
                     /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     3692..3711
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3930..4034)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4102..4120
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctagaggagg aaaattaatg tcagaactta tcaaggaaaa tatgcacatg aaattgtaca
       61 tggaaggaac tgtcaataat caccacttta aatgtacctc agaaggagaa ggaaaaccat
      121 atgaaggtac tcaaaccatg cgtattaagg ttgtcgaagg tggaccactt ccttttgctt
      181 ttgatattct tgcaacatct tttatgtacg gatcacgtac ctttatcaat catacacaag
      241 gtatcccaga tttctttaaa cagtcttttc ctgaaggatt tacatgggaa cgtgttacaa
      301 cttatgaaga tggtggagtc ttgacagcta ctcaagatac atctttgcaa gatggttgtt
      361 tgatctacaa tgtcaagatc cgtggagtaa attttccatc taatggtcct gttatgcaga
      421 aaaagactct tggatgggaa gccaataccg aaatgttgta tcctgctgat ggtggacttg
      481 aaggtcgttc agatatggca ttgaaacttg ttggtggagg tcatttgatc tgtaatttta
      541 agaccacata ccgtagtaaa aagccagcaa agaatcttaa gatgcctggt gtatactacg
      601 tcgatcatcg tttggaacgt atcaaagaag ccgataagga aacttatgtt gaacaacacg
      661 aagtcgccgt agcacgttac tgtgatttgc ctagtaaatt gggacataaa taataataag
      721 gatcctaact cgagaaagaa ttcatgacta gtcaaggtcg gcaattctgc agtactagga
      781 cgccgccaag ccagcttaaa cccagctcaa tgagctgggt tttttgtttg ttaaaaatga
      841 agaagaaact gtgaagcgta tttatagcaa agcactcaaa agtttacctt atgggtgctt
      901 ttttcgtgct tttttgaaaa gacaaaaaaa agaaccttgc caagcaagat tcttttgata
      961 gcgctatcgc tgagcgccgg tcgctaccat taccagttgg tctggtgtca aaaataataa
     1021 taaccgggca ggccatgtct gcccgtattt cgcgtaagga aatccattat gtactatttc
     1081 tggtgatgaa atcaacgtaa catttaaagc tgtcaaagcc aaagtcatga gatggcgtat
     1141 ggagcgtaaa gctgacaaga gcggtgttgc gatgattgag atgaccttcc ttgcaccaag
     1201 tgaattgcct caagaaagca ctcaatcaaa gattcttgta gatggaaaag aacttgctga
     1261 tttcgctgaa aatcgtcaag actatcaaat tacctataaa ggtcaacggc caaaagtctc
     1321 agttgaagaa aacaatcaag tagcttcaac tgtggtagat agtggagaag atagccttcc
     1381 agtacttgtt cgcctcgttt cagaaagtgg aaaacaagtc aaggaatacc gtatccagtt
     1441 gactaaggaa aaaccagttt ctgctgtaca agaagatctt ccaaaactcg aatttgttga
     1501 aaaagatttg gcctacaaga cagttgagaa aaaagattca acactgtatc taggtgaaac
     1561 tcgtgtagaa caagaaggaa aagttggaaa agaacgtatc tttacagtga ttaatcctga
     1621 tggaagtaag gaagaaaaac tccgtgaagt ggtagaagtt ccgacagacc gcatcgtctt
     1681 ggttggaacc aaaccagtag ctcaagaagc taaaaaacca caagtgtcag aaaaagcaga
     1741 tacaaaacca attgattcaa gtgaagctga tcaaactaat aaagcccagt taccaaatac
     1801 aggtagtgcg gcaagccaag cagcagtagc agcaggttta gcctgcctcg cgcgtttcgg
     1861 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     1921 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     1981 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg
     2041 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     2101 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     2161 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2221 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2281 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2341 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2401 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2461 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2521 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2581 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2641 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2701 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     2761 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2821 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2881 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2941 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3001 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3061 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3121 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3181 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3241 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3301 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3361 ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg
     3421 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3481 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3541 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3601 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3661 ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta
     3721 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3781 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3841 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     3901 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3961 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4021 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     4081 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct tcaagaatta
     4141 attccttctt aacgccccaa gttcatcacc aatgacatca actcacatga actacatgat
     4201 gaacccagtt atcatggttt tggataagat ttttgaaaaa ttcttcccag gccttgataa
     4261 atatgacttt gatgctgcta aattgaacaa gaaaatcggt ttctggggat ctaaattctt
     4321 catcggtttc atccttggta tcgttatcgg tattatggga actccacatc caattgcagg
     4381 tgttgcagat gcagataaat ggcgtcttgt tatcaaagga tggttgtctc ttggtttgac
     4441 tgccggtgta tctttggaac tcttctcact tatcggttca tggttcatcg cagccgtaga
     4501 accactatca caaggtatta caaacgttgc tactaaacgt cttcaaggac gtaaattcaa
     4561 tatcggtctt gactggccat tcatcgctgg tcgtgctgaa atctgggctt gtgccaacgt
     4621 acttgcacca atcatgttga ttgaagcagt gcttctttca aaagttggaa atggtatctt
     4681 gccacttgca ggtatcatcg ctatgggtgt tactccagct ctcttggttg taactcgtgg
     4741 taaattgctc cgtatgatta tcttcggaac actcttgttg ccactcttcc ttctttcagg
     4801 tacacttatt gcaccatttg caacagaact tgctaaaggt gtaggtgcct tcccagaagg
     4861 tgtgagccaa actcaattga ttactcactc tactcttgaa ggaccaatcg aaaaacttct
     4921 tggttggaca attggtaaca ctacaactgg tgatatcaaa gcaatccttg gtgcagtagc
     4981 cttccttgta ttctatatcg gtatctttgc ttggtacaga aaacaaatga tcaaacgtaa
     5041 cgaagagtac gcagcaaaag caaaataatg cgctccgcta gctttacaga caaagaacta
     5101 tccttaatgg aagcactagc aaaaggtatc aatgaagcaa gaaatattga aggctagtca
     5161 gtaaaattca gataaacaaa aagagccgat aagatgagag tttcatttct caaattatcg
     5221 gctctgcgtc ttggcgtctg gctctttgat tgttattata ttcatttttt gaacattaat
     5281 taaagtttcg ttcttacatt tggggcagta taaagggaaa tttttaagta tagtatccac
     5341 tcgtattcgt agtcttgttt tatttccaca aataggacac aatatccact tgtagtttat
     5401 aataacaatc tcctcctttc cactttaatt caaatctata ttaaagaata tttcatctta
     5461 tttaataaga aaccatattt atataacaac ataaaacgca ctaagttatt ttattgaaca
     5521 tatatcttac tttatctatc tgactattta gacgacgggt ctggcaaaca ggttcgccag
     5581 tggtaacctg atatcctttt agctctgcta aacaaacact aagcccattt gtaaaaaaag
     5641 ttaaatcatt gcgataatct tgaatacatc gagcaggaat ttctccaata ataatgacct
     5701 cattattttt cagttgagta tttacgatat ttgcacaata tttgggagca tcgttatatg
     5761 cccgtgaaag atattcctgt ggtgcataaa ctttaaaact aagatatggc tctaacaatt
     5821 ctgttccagc ttttctaaag gcttgctcca atacaatagg agcaagcatc cgaaaatctg
     5881 ctggggtact aacagggcta tagtataagc catacttaaa acagatttta cagtccgtca
     5941 cattccaacc atacaatcct tgttcacaac catagcgtat cccttccata actgcatttt
     6001 gaaatgattg atttaagtat ccaagagaaa ccgagctctc atactgcatt ccacttccca
     6061 acggaagcgg tgatacagat aaaccaatgg aagcccagaa aggatttggc ggcacttcga
     6121 tgtgaatggt atattctgca ttttttaacg gtctctccat ataaatgact gtaggctctt
     6181 ttagttctat ctccacatga tacttttctt gcaacagtgc actaatcact tccatttgta
     6241 ctttccctaa gaaagaaagt ataatttcat gtgtcgtaga atccacgtaa tatcgtagaa
     6301 gcggatcact atctgagatt tccaaaaggg catcaagcaa catttctctc tgttcaggtt
     6361 tactcggttc aacagttgtt tgtagtagag ggtgcggatt ttcaatcttt tttctctgtg
     6421 gcaatagttt tgtatctcca agaacactat ttaacttcaa aaactcattt tgcaaaataa
     6481 caatttctcc agaataagct ctatcaatct tacataattc accatttatt gaagtataca
     6541 tttctgtaac ttttattttt tctttttctg atactctaac cgaatctcgt aaatgtagta
     6601 ctccactata aaggcgtata tatgcaagac gttgtctttt ttttgtatat tcaattttga
     6661 aaacatttcc gcaaagttca gacggacctc gatgtgttga tgaataaaat ttattagtaa
     6721 taacttctat aaggttatca atccctatat tactttttgc acttccatga taaagaggga
     6781 acagagaaca attctgaaat cttatgcttt cctcttgttc gagttccaat gcttctaatg
     6841 atttaccgga catatatttc tctaaaaggt catcgtttcc ctctattacc gtatcccatt
     6901 gttcagattc ggtaaagttc gtcacacaca cattaggata cagttctacc ttctgtttga
     6961 ttacaatttc ggcagaaagt ttctctttaa tatcctgata aaccgttgat aaatcaattc
     7021 cattttggtc aatcttattg ataaaaaaga ttgtgggaat ccccattttc ctaagtgcat
     7081 gaaataatat acgagtttgt gcttgtacgc catcttttgc agaaatcagt agaattgccc
     7141 catctaaaac tgataatgaa cgatatactt ctgctaagaa atccatatgt cctggcgtgt
     7201 ctatgatgtt caccttcgta ttttcccact gaaaagaggt tattcctgtc tgaattgtaa
     7261 ttcctctctg acgttctaaa agcgtattat ccgtcctcgt tgtacctttg tccacgcttc
     7321 ctaattctgt aatcgctcca ctgttatata ataagctttc tgttaaggta gtttttcctg
     7381 catcaacatg agctaaaact ccaatattaa taattttcat gtgattttcc tccattcaaa
     7441 agcccaaaag ggcataaaaa tcccagtgat aaatactctt atcactggga tttttatgca
     7501 taaccatagg catacaaagc atacagatat tctccggata ctttagaatc acatgataaa
     7561 ggtattctta aactgggtac aaaaaactaa gccctcctaa aaaaggacat ccaattattt
     7621 gttcccgcta tcaaattgac agtttattta agaatacctt gccgcatatt tattaactcc
     7681 ttttaaatag atacttaaat tatagcacgt aagagcatat ttgtaaagga atctccaatt
     7741 ttttatcaaa aagagtacat gattacaaag tatctgtaat catgtaccaa tatttgttat
     7801 tttacaatct tccaattatc ttgtttttct aatatcaact caaattgcga gatttgggtt
     7861 gcctttgttt cttgatctaa atatttcaca gcgacattca cgattacttg gctagcgcta
     7921 tatgcgttga tgcaatttct atgcgcaccc gttctcggag cactgtccga ccgctttggc
     7981 cgccgcccag tcctgctcgc ttcgctactt ggagccacta tcgactacgc gatcatggcg
     8041 accacacccg tcctgtggat ctatcgatgc atgcagccgg taccatcaca agcactttgg
     8101 gacgttctcc cttagtgctt ttttgatttc tcataggccg gcctgttagt catatggaca
     8161 cttaaggcaa attgttcaga actgaataaa gctgacgttt tgcttctatc ctttctttga
     8221 gttttagtgg ataatgataa tgaacaaggt gttcataaat ctattataac aagaattcca
     8281 atcccgggta aagcttccag cggccgcata ggatccattg atatcatatt
//
