LOCUS       sequence-245474-        7462 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli-streptococcal shuttle vector that expresses rfp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7462)
  AUTHORS   Shields RC, Kaspar JR, Lee K, Underhill SAM, Burne RA
  TITLE     Fluorescence tools adapted for real-time monitoring of the behaviors
            of Streptococcus species.
  JOURNAL   Appl Environ Microbiol. 2019 May 17. pii: AEM.00620-19. doi: 
            10.1128/AEM.00620-19.
  PUBMED    31101614
REFERENCE   2  (bases 1 to 7462)
  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. 2019 May 17. pii: AEM.00620-19. doi: 
            10.1128/AEM.00620-19."
COMMENT     Sequence Label: pDL278_P23-DsRed-Express2
FEATURES             Location/Qualifiers
     source          1..7462
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..24,25..31)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    39..55
                     /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)."
     primer_bind     44..66
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     63..79
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             244..921
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(424..444)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     642..661
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     836..859
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(989..1006)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(989..1005)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(998..1020)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      1479..1565
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1657..1684
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      1977..2063
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2155..2182
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      6401..6989
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6890..6909
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7143..7160
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7427..7448
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
       61 cacaggaaac agctatgacc atgattacgc caagctttcg aaaagccctg acaaccctcg
      121 ttcctaaaaa ggaataagcg tttggtcagt aaataataga aataaaaaat cagacctaag
      181 actgatgaca aaaagagcaa attttgataa aatagtatta attactagtt aaggaggaca
      241 aacatggata gcactgagaa cgtcatcaag cccttcatgc gcttcaaggt gcacatggag
      301 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggcaa gccctacgag
      361 ggcacccaga ccgccaagct gcaggtgacc aagggcggcc ccctgccctt cgcctgggac
      421 atcctgtccc cccagttcca gtacggctcc aaggtgtacg tgaagcaccc cgccgacatc
      481 cccgactaca agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
      541 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg caccttcatc
      601 taccacgtga agttcatcgg cgtgaacttc ccctccgacg gccccgtaat gcagaagaag
      661 actctgggct gggagccctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc
      721 gagatccaca aggcgctgaa gctgaagggc ggcggccact acctggtgga gttcaagtca
      781 atctacatgg ccaagaagcc cgtgaagctg cccggctact actacgtgga ctccaagctg
      841 gacatcacct cccacaacga ggactacacc gtggtggagc agtacgagcg cgccgaggcc
      901 cgccaccacc tgttccagta gctcggtacc aaattccaga aaagaggcct cccgaaaggg
      961 gcgccttttt tcgttttggt ccgaattcac tggccgtcgt tttacaacgt cgtgactggg
     1021 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     1081 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     1141 aatgccggga attattcacc actccaagaa ttggagccaa tcaattcttg cggagaactg
     1201 tgaatgcgca aaccaaccct tggcagaaca tatccatcgc gtccgccatc tccagcagcc
     1261 gcacgcggcg catctcggct gttttggcgg atgagagaag attttcagcc tgatacagat
     1321 taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt
     1381 ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt
     1441 ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
     1501 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     1561 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
     1621 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
     1681 tttttgcgtt tctacaaact cttcctgtcg tcatatctac aagccatccc cccacagata
     1741 cggtaaacta gcctcgtttt tgcatcagga aagcagctgt tttggcggat gagagaagat
     1801 tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc
     1861 tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg
     1921 tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa
     1981 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     2041 acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc
     2101 ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg
     2161 ccatcctgac ggatggcctt tttgcgtttc tacaaactct tcctgtcgtc atatctacaa
     2221 gccatccccc cacagatacg gtaaactagc ctcgtttttg catcaggaaa gcagtcgggc
     2281 agcgttggga ttctgagacc tctgagaggc tctcagagct atctaaagct gagggatata
     2341 taaatacctt agaaaattat tcgaagagct tagaagcgaa aatagagcgt ttagagcgcg
     2401 aggggctgaa attagaaaaa ctaaaaacac aaatagctga cctaaaaatc atgtctgaga
     2461 aagaactagc ggctattacc cctaaaaaag gcgtgttcgg taaagaatat gtggaattga
     2521 ctaaagagca gtttgaagaa tttaaagggc tgatataccg aagcagaaac cttgttcatc
     2581 aaaaagagct agagaatgag caattaaggc ggatagtgcc tctgagacgc tctaaaaacg
     2641 gttttgaagc gagtttggaa cgagctaaag aaaaaagtaa gggagagagc atagagcgtc
     2701 ttaggaacga aaatagagcg cttagaagtg aaaactcagt tttgagacag caaaatgaca
     2761 aaatgctagg aaaactgaaa gagtttatgc cagataaagc cttaaaaaat tttatatcag
     2821 agttaagagc tattcagcca atcgtaaggg tagttaaacg agtgattgaa aaagggctag
     2881 gcctttgagc gatttatgcc gtgaaagcta attgacaata agcaagggca aagtacgcta
     2941 ggacgtgacg agccgaaagg ctttagcgtt tcgagccgac acggacaaag gacgtccgcc
     3001 cttggttact tgttgtcaat tagaccatgg aataaagtaa gcggacatgg tataatagct
     3061 aggtcgcaac gttctttcgc taagttacga acttagattg gaggtgagcg ctgtgaagac
     3121 tttcctagaa cttgttttga taccttttgt ggttggcgtt gctgcagagg taagtgctga
     3181 ttggttgatt cggtatgttc gaaacaaacg cgacaaaaat taacctgagt tctttttgag
     3241 gacaaaaaag aaagacagta gttccagcta ctgttttttt tgcgttgtgc tattcgtttc
     3301 ctagaacttc tagcgttaaa attattatac cacgtttgga tttagaaagt caaaatttga
     3361 ggttttaggg gtgaattttt cgtgaaacga aaaagagggc tgaaaagccc ttaaaaaccc
     3421 atgcgtagca agggtttttt cttatcttga taaaataaag tttataattt aagtgtccaa
     3481 tttaaaataa taaacttagg agaattgcag gaactttttt atatactcaa aaaaattttt
     3541 ttgcaagaaa attataacat gacaggtact gaaaatcaag tctttaagga ctattctaag
     3601 aatggcaaag atagaaagtg gcgagaacgc aagttaaaaa atattgagct tgctggtcgt
     3661 ttagagcgtt taggatatcg ttcgtttgaa cgggtctatc aatgtgccga agtgttgaag
     3721 tttatcgaac aacaagacgg cacgaaaaaa ctctatcagt cttatttttg caaaaataag
     3781 ctctgcccaa tttgtaactg gagacggtca atgaagtatg cttatcaagc tgaattagtg
     3841 gtaaatgaag caatgaaacg ctatccaaaa ggtcgctttc tctttttgac cttgacgatt
     3901 aagaacatta gtggtgaaaa attaaataaa tcaatttcag aaatagggcg agcttttaat
     3961 cgtcttatga aatacaagaa agtcgataaa aatgttattg gctatttgcg agccactgag
     4021 gtaacttatt caactgagca tgagaattat caccctcatt tgcatgtatt gttatttgtg
     4081 aaatctagct attttactgg aaataataca aattacatta gccaagaaga atggacgaaa
     4141 ctatgggcta aggcgatgaa attggattat acacctgtag ttgatattcg aaccgtcaaa
     4201 gctcataaac gtaaaaactt gaagtcagct attatcgaaa cagctaaata tcctgttaag
     4261 ccttttgatg tagatacaga agatgtgaca ttattctctg aaatggtcaa agaacggata
     4321 acagaagatt taacgaatgg tttgcaccga aaaaggcaga ttggttttgg aaagttgttc
     4381 aagaaaatca aggcggagtt agctcttgat gatgtcgaag aagggaatct tgttcagacc
     4441 ggagcagaag aatctgcaga aagtactggt cgtgaaattg ttgccttttg gaattgggat
     4501 agaaagaatt attttgtgag gtagctgatg agtaaaacag tatcagaatt agctcaagaa
     4561 ttgggagtta gtaggcagta tcttaatcgg attttatcgc aaaataatct cggtcgaaaa
     4621 aaagggaata aaaaagtagt ttccgatatg gacgagaaag ttatgcaagg gtttattgtt
     4681 ttctaaaatc tgattaccaa ttagaatgaa tatttcccaa atattaaata ataaaacaaa
     4741 aaaattgaaa aaagtgtttc caccattttt tcaatttttt tataattttt ttaatctgtt
     4801 atttaaatag tttatagtta aatttacatt ttcattagtc cattcaatat tctctccaag
     4861 ataactacga actgctaaca aaattctctc cctatgttct aatggagaag attcagccac
     4921 tgcatttccc gcaatatctt ttggtatgat tttacccgtg tccatagtta aaatcatacg
     4981 gcataaagtt aatatagagt tggtttcatc atcctgataa ttatctatta attcctctga
     5041 cgaatccata atggctcttc tcacatcaga aaatggaata tcaggtagta attcctctaa
     5101 gtcataattt ccgtatattc ttttattttt tcgttttgct tggtaaagca ttatggttaa
     5161 atctgaattt aattccttct gaggaatgta tccttgttca taaagctctt gtaaccattc
     5221 tccataaata aattcttgtt tgggaggatg attccacggt accatttctt gctgaataat
     5281 aattgttaat tcaatatatc gtaagttgct tttatctcct attttttttg aaataggtct
     5341 aattttttgt ataagtattt ctttactttg atctgtcaat ggttcagata cgacgactaa
     5401 aaagtcaaga tcactatttg gttttagtcc actctcaact cctgatccaa acatgtaagt
     5461 accaataagg ttatttttta aatgtttccg aagtattttt ttcactttat taatttgttc
     5521 gtatgtattc aaatatatcc tcctcactat tttgattagt acctatttta tatccatagt
     5581 tgttaattaa ataaacttaa tttagtttat ttatagattt cattggcttc taaatttttt
     5641 atctagataa taattatttt agttaatttt attctagatt atatatgata tgatctttca
     5701 tttccataaa actaaagtaa gtgtaaacct attcattgtt ttaaaaatat ctcttgccag
     5761 tcacgttacg ttattagtta tagttattat aacatgtatt cacgaacgaa aaatcttcta
     5821 tataaaagat atattatctt atcagtattg tcaatatatt caaggcaatc tgcctcctca
     5881 tcctcttcat cctcttcgtc ttggtagctt tttaaatatg gcgcttcata gagtaattct
     5941 gtaaaggtcc aattctcgtt ttcatacctc ggtataatct tacctatcac ctcaaatggt
     6001 tcgctgggtt tatcggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6061 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6121 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6181 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     6241 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6301 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6361 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6421 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6481 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6541 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6601 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6661 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6721 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6781 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6841 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6901 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6961 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7021 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     7081 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     7141 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     7201 cgcgttggcc gattcattaa tgcagaaata cgtttcccat ttagccgtca tttcagcact
     7261 cgttaaaagg tgctgacttt caactgcctg gcacaataat tttccttttt cagttacaac
     7321 aagctccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     7381 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     7441 tcactcatta ggcaccccag gc
//
