LOCUS       sequence-193485-        7362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of RFP in Lactobacillus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7362)
  AUTHORS   Bao S, Zhu L, Zhuang Q, Wang L, Xu PX, Itoh K, Holzman IR, Lin J
  TITLE     Distribution dynamics of recombinant Lactobacillus in the 
            gastrointestinal tract of neonatal rats.
  JOURNAL   PLoS One. 2013;8(3):e60007. doi: 10.1371/journal.pone.0060007. Epub 
            2013 Mar 27.
  PUBMED    23544119
REFERENCE   2  (bases 1 to 7362)
  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: "PLoS One.";
            date: "2013"; volume: "8(3):e60007. doi"; pages: " 
            10.1371/journal.pone.0060007. Epub 2013 Mar 27"
COMMENT     Sequence Label: pLEM415-ldhL-mRFP1
FEATURES             Location/Qualifiers
     source          1..7362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     46..68
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     60..77
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     61..77
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     87..106
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        87..105
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(442..450,451..465)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             472..1149
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             472..1149
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(602..620)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     870..889
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1064..1087
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(4582..4598)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4582..4598)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4635..4655)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4635..4653)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4674..4690)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4674..4690)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4687..4709)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4698..4714
                     /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)."
     promoter        complement(join(4722..4728,4729..4746,4747..4752))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4767..4788
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4905..4922)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5076..5664)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5156..5175)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5835..6626,6627..6695))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6458..6477
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6696..6800)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6826..7281)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6963..6984)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7175..7194
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
       61 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtaccg
      121 ggcccactga gaagttgctc tccccagtcg acgtttgtta ttgctgttta ttataccata
      181 cccgtttttc tttctatatt agggaataaa tagttttctg aaaatttaaa caaaattagc
      241 gaactttcat gcaaactatg ctatactggt tttcgaaatg aagaattgtt ttacagaaag
      301 gacgtcatta tattggcaaa cattgaaaaa gatcaccaaa aagttatcct cgtcggcgaa
      361 agcttgcatg cctgcaggtc gactctagag gatccccggg taccggtaga aaaaatgact
      421 ggtggacagc aaatgggtcg ggatctgtac gacgatgacg ataagcctag gatggcctcc
      481 tccgaggacg tcatcaagga gttcatgcgc ttcaaggtgc gcatggaggg ctccgtgaac
      541 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
      601 gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtcccct
      661 cagttccagt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
      721 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
      781 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
      841 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
      901 gaggcctcca ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagatg
      961 aggctgaagc tgaaggacgg cggccactac gacgccgagg tcaagaccac ctacatggcc
     1021 aagaagcccg tgcagctgcc cggcgcctac aagaccgaca tcaagctgga catcacctcc
     1081 cacaacgagg actacaccat cgtggaacag tacgagcgcg ccgagggccg ccactccacc
     1141 ggcgcctaag ctctacaaat aaatcgataa gcttgatatc gaattcctgc agctaattgg
     1201 gtgttcatga cgaccaataa tagtatcagg agtccaacct gccttgattt gcgcatggat
     1261 atagttaact tcgatagttg gcagttgggt ctgcttccga ccacaacgac gcttatgtcg
     1321 ctgataagtc tgaagatatt ggtcgatggt tttaccgtcg ttgaggaaac gataaacacg
     1381 atagatggtt tcttgactac gctgaagtaa tttagcagcc cgataagctt tagtaccttg
     1441 ataccaaaaa tcagctatga gagttaattc acgtgtggta agatgtttat aggtcatttg
     1501 tgattgcctt tcttttgatt aaggatattc aaaagtctat cacaaatggc tttttatttt
     1561 ttctaactta attttacaaa cgacgaaaca aaaaaagagg ctgattaagt tcagtctctt
     1621 ttttaattaa ttgtgtgatt cttcaatttc agccaactta gatgaagctt gatagatcag
     1681 tgattctgcg tcactcaata gagataatat atcatcatat gagccatcat ctgcttggct
     1741 caaaattcgc tttttaaaat cctctaaatt atcagccaac tgctcgctaa tcgtcccata
     1801 aatttttgac tgttcaataa ttgcatcaat gccttttaca tctcttaact ccataacttt
     1861 cactcccaac ttacataatc gctaacacta caattccact tatacattac cttttttaca
     1921 atttcagaat catcattagt ggcacttaca ttaatcagat catcttcatt ttcatcaaat
     1981 aacaaactat gtctaatatc ttttaacaat ccgccaaaac tcaactgcct agatccctgc
     2041 aatccctttt ctagttcgtt taaaataacc aaatcagatt cttgatcatt cgttatataa
     2101 tcagaatttt ttacttggta cttagcaact tctttagctg aatcaactaa tgccgattga
     2161 ccatgtttac ttttagttct tatcttacga atatcaataa taggatcata atcgagtttt
     2221 ctagcacgac gccaaagttt tatccaatca tcttttgtca aataattttt tgaattaaaa
     2281 taacttgtct tcattaacaa taataaatgc atatgttgat gaaacgtata attatctcga
     2341 ttgactgtaa tttcagaaga tcgtacatag ccaatcatat ttttagctac ttttttatac
     2401 tgaattaagc gtctaactga actattcata ttttttaaat ttatttttaa ttctccaatt
     2461 tcggaatttt cctctgtaag agttaaaaat aaataaatcg catttggatt tttaacatgc
     2521 gctatatcta aaatttgttg caactcgtaa ctatttttta aagatcttct ccaagaacac
     2581 aatggacata acctagagtg acagaaccac gtttgataca gtttcattcc atccttgcct
     2641 atagcaaatt gtaatacttc accacaatct ttaacacgat tagctttctt aaattttaaa
     2701 acctccaaat attgcgcata agtaagatta gctagtttct tttctcgcca tggtcttact
     2761 ttgccactat atttagaggt atctcttagt ctttcacctg tatacactta taaatacctc
     2821 tttactgaaa ttcatgcagt aaagagagtt agctatacta attgatataa taataattaa
     2881 ctactataat agataattga taatattaga taactagctc atgcacctac cgatcgccaa
     2941 atctaaaggg actgcatgag ttttttattt acgaataaat tatacctcta ttctattagt
     3001 aatcaaaatt tctacaaaca ttgatatatc aacgctttaa cagcatttta aaaaatgacg
     3061 ttgtttctat aagtatcaag ataagtagac acgcgattaa aatcgcgtgt cttagggcgc
     3121 tgtcgcccct aaaaccccga ccaagcgtcg gacgcttgac ccccgctagc ccttgctaga
     3181 cggcgggcga aaaataaatt tttcaccaaa taaaagggtc actcactatt attcgtcaag
     3241 ggttgccttg cttccctccg gtcgcaagcg tgtgcccctt gacgaatggc atctaataga
     3301 gggctacgcc accagcccac aattagcaaa aaggcaacat tcctccgcac ggcgggttat
     3361 gttgcctttt tgctttttaa aaattaaact taaggaatat caagcttcat agcgcctaaa
     3421 atatagggag ttgtttcctc ccgttaaata atagataact attaaaaata gacaatactt
     3481 gctcataagt aatggtactt aaattgttta ctttggcgtg tttcattgct tgatgaaact
     3541 gatttttagt aaacagttga cgatattctc gattgaccca ttttgaaaca aagtacttat
     3601 atagcttcca atatttatct ggaacatctg tggtatggcg ggtaagtttt attaagacac
     3661 tgtttacttt tggtttagga tgaaagcatt ccgctggcag cttaagcaat tgctgaatcg
     3721 agacttgagt gtgcaagagc aaccctagtg ttcggtgaat atccaaggta cgcttgtaga
     3781 atccttcttc aacaatcaga tagatgtcag acgcacggct ttcaaaaacc acttttttaa
     3841 taatttgtgt gcttaaatgg taaggaatat tcccaacaat tttatacctc tgtttgttag
     3901 ggaattgaaa ctgtagaata tcttggtgaa ttaaagtgac acgagtattc agttttaatt
     3961 tttctgacga taagttgaat agatgactgt ctaattcaat agacgttacc tgtttactta
     4021 ttttagccag tttcgtcgtt aaatgccctt tacctgttcc aatttcgtaa acggtatcgg
     4081 tttcttttaa attcaattgt tttattattt ggttgagtac tttttcactc gttaaaaagt
     4141 tttgagaata ttttatattt ttgttcatgt aatctctcct gaagtgatta catctgtaaa
     4201 taaatacaga agttaaacga tttgtttgta attttagtta tctgtttaaa aagtcataag
     4261 attagtcact ggtaggaatt aatctaacgt atttatttat ctgcgtaatc actgttttta
     4321 gtctgtttca aaacagtaga tgttttatct acattacgca tttggaatac caacatgacg
     4381 aatccctcct tcttaattac aaatttttag catctaattt aacttcaatt cctattatac
     4441 aaaattttta aacgaatttt ttaaaaatgt attgagcttc tgccaattaa ttttaggaac
     4501 agcaatgtga ctaatcccta tcccatctcg atatatcagt ccttgaccaa cattgttgtt
     4561 aagccaatct gcagcccggg ggatccacta gttctagagc ggccgccacc gcggtggagc
     4621 tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     4681 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4741 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4801 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4861 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4921 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4981 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5041 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5101 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5161 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5221 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5281 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5341 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5401 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5461 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5521 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5581 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5641 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5701 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5761 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5821 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5881 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5941 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6001 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6061 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6121 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6181 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6241 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6301 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6361 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6421 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6481 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6541 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6601 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6661 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6721 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6781 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     6841 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     6901 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     6961 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     7021 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     7081 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     7141 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     7201 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     7261 atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc
     7321 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa ag
//
