LOCUS       sequence-11337-e        8509 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8509)
  AUTHORS   Lizier M, Sarra PG, Cauda R, Lucchini F
  TITLE     Comparison of expression vectors in Lactobacillus reuteri strains.
  JOURNAL   FEMS Microbiol Lett. 2010 Jul 1. 308(1):8-15.
  PUBMED    20455948
REFERENCE   2  (bases 1 to 8509)
  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: "FEMS 
            Microbiol Lett. 2010 Jul 1. 308(1):8-15."
COMMENT     Sequence Label: pTRKH3-ldhGFP
FEATURES             Location/Qualifiers
     source          1..8509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5757..5776
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5843..5945)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(6337..6354)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6471..7016)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(6551..6570)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(7128..7133,7134..7150,7151..7156)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             complement(7499..8215)
                     /codon_start=1
                     /gene="mgfp5"
                     /product="GFP with folding enhancement mutations"
                     /label=mgfp5
                     /note="suitable for expression in plants due to removal of 
                     a cryptic intron"
                     /translation="MSKGEELFTGVVPILVELDGDVNGYKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKTRHNIEDGGVQLADHYQQNTPIGDDPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(7542..7562)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     8157..8185
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
       61 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
      121 ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg
      181 cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc
      241 cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct
      301 gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct
      361 tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga
      421 tgacgaccat cagggacagc ttcaaggatc gctcgcggct cttaccagcc taacttcgat
      481 cactggaccg ctgatcgtca cggcgattta tgccgcctcg gcgagcacat ggaacgggtt
      541 ggcatggatt gtaggcgccg ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc
      601 atggagccgg gccacctcga cctgaatgga agccggcggc acctcgctaa cggattcacc
      661 actccaagaa ttggagccaa tcaattcttg cggagaactg tgaatgcgca aaccaaccct
      721 tggcagaaca tatccatcgc gtccgccatc tccagcagcc gcacgcggcg catctcgagt
      781 ctagaatcga tacgattttg aagtggcaac agataaaaaa aagcagttta aaattgttgc
      841 tgaactttta aaacaagcaa atacaatcat tgtcgcaaca gatagcgaca gagaaggcga
      901 aaacattgcc tggtcgatca ttcataaagc aaatgccttt tctaaagata aaacgtataa
      961 aagactatgg atcaatagtt tagaaaaaga tgtgatccgt agcggttttc aaaatttgca
     1021 accaggaatg aattactatc ccttttatca agaagcgcac aaaaagaaaa acgaaatgat
     1081 acaccaatca gtgcaaaaaa agatataatg ggagataaga cggttcgtgt tcgtgctgac
     1141 ttgcaccata tcataaaaat cgaaacagca aagaatggcg gaaacgtaaa agaagttatg
     1201 gaaataagac ttagaagcaa acttaagagt gtgttgatag tgcagtatct taaaattttg
     1261 tataatagga attgaagtta aattagatgc taaaaatttg taattaagaa ggagtgatta
     1321 catgaacaaa aatataaaat attctcaaaa ctttttaacg agtgaaaaag tactcaacca
     1381 aataataaaa caattgaatt taaaagaaac cgataccgtt tacgaaattg gaacaggtaa
     1441 agggcattta acgacgaaac tggctaaaat aagtaaacag gtaacgtcta ttgaattaga
     1501 cagtcatcta ttcaacttat cgtcagaaaa attaaaactg aatactcgtg tcactttaat
     1561 tcaccaagat attctacagt ttcaattccc taacaaacag aggtataaaa ttgttgggag
     1621 tattccttac catttaagca cacaaattat taaaaaagtg gtttttgaaa gccatgcgtc
     1681 tgacatctat ctgattgttg aagaaggatt ctacaagcgt accttggata ttcaccgaac
     1741 actagggttg ctcttgcaca ctcaagtctc gattcagcaa ttgcttaagc tgccagcgga
     1801 atgctttcat cctaaaccaa aagtaaacag tgtcttaata aaacttaccc gccataccac
     1861 agatgttcca gataaatatt ggaagctata tacgtacttt gtttcaaaat gggtcaatcg
     1921 agaatatcgt caactgttta ctaaaaatca gtttcatcaa gcaatgaaac acgccaaagt
     1981 aaacaattta agtaccgtta cttatgagca agtattgtct atttttaata gttatctatt
     2041 atttaacggg aggaaataat tctatgagtc gcttttgtaa atttggaaag ttacacgtta
     2101 ctaaagggaa tgtagataaa ttattaggta tactactgac agcttccaag gagctaaaga
     2161 ggtccctagc gctcttatca tggggaagct cggatcatat gcaagacaaa ataaactcgc
     2221 aacagcactt ggagaaatgg gacgaatcga gaaaaccctc tttacgctgg attacatatc
     2281 taataaagcc gtaaggagac gggttcaaaa aggtttaaat aaaggagaag caatcaatgc
     2341 attagctaga actatatttt ttggacaacg tggagaattt agagaacgtg ctctccaaga
     2401 ccagttacaa agagctagtg cactaaacat aattattaac gctataagtg tgtggaacac
     2461 tgtatatatg gaaaaagccg tagaagaatt aaaagcaaga ggagaattta gagaagattt
     2521 aatgccatat gcgtggccgt taggatggga acatatcaat tttcttggag aatacaaatt
     2581 tgaaggatta catgacactg ggcaaatgaa tttacgtcct ttacgtataa aagagccgtt
     2641 ttattcttaa tataacggct ctttttatag aaaaaatcct tagcgtggtt tttttccgaa
     2701 atgctggcgg taccccaaga attagaaatg agtagatcaa attattcacg aatagaatca
     2761 ggaaaatcag atccaaccat aaaaacacta gaacaaattg caaagttaac taactcaacg
     2821 ctagtagtgg atttaatccc aaatgagcca acagaaccag agccagaaac agaatcagaa
     2881 caagtaacat tggatttaga aatggaagaa gaaaaaagca atgacttcgt gtgaataatg
     2941 cacgaaatcg ttgcttattt ttttttaaaa gcggtatact agatataacg aaacaacgaa
     3001 ctgaatagaa acgaaaaaag agccatgaca catttataaa atgtttgacg acattttata
     3061 aatgcatagc ccgataagat tgccaaacca acgcttatca gttagtcaga tgaactcttc
     3121 cctcgtaaga agttatttaa ttaactttgt ttgaagacgg tatataaccg tactatcatt
     3181 atatagggaa atcagagagt tttcaagtat ctaagctact gaatttaaga attgttaagc
     3241 aatcaatcgg aaatcgtttg attgcttttt ttgtattcat ttatagaagg tggagtttgt
     3301 atgaatcatg atgaatgtaa aacttatata aaaaatagtt tattggagat aagaaaatta
     3361 gcaaatatct atacactaga aacgtttaag aaagagttag aaaagagaaa tatctactta
     3421 gaaacaaaat cagataagta tttttcttcg gagggggaag attatatata taagttaata
     3481 gaaaataaca aaataattta ttcgattagt ggaaaaaaat tgacttataa aggaaaaaaa
     3541 tctttttcaa aacatgcaat attgaaacag ttgaatgaaa aagcaaacca agttaattaa
     3601 acaacctatt ttataggatt tataggaaag gagaacagct gaatgaatat cccttttgtt
     3661 gtagaaactg tgcttcatga cggcttgtta aagtacaaat ttaaaaatag taaaattcgc
     3721 tcaatcacta ccaagccagg taaaagcaaa ggggctattt ttgcgtatcg ctcaaaatca
     3781 agcatgattg gcggtcgtgg tgttgttctg acttccgagg aagcgattca agaaaatcaa
     3841 gatacattta cacattggac acccaacgtt tatcgttatg gaacgtatgc agacgaaaac
     3901 cgttcataca cgaaaggaca ttctgaaaac aatttaagac aaatcaatac cttctttatt
     3961 gattttgata ttcacacggc aaaagaaact atttcagcaa gcgatatttt aacaaccgct
     4021 attgatttag gttttatgcc tactatgatt atcaaatctg ataaaggtta tcaagcatat
     4081 tttgttttag aaacgccagt ctatgtgact tcaaaatcag aatttaaatc tgtcaaagca
     4141 gccaaaataa tttcgcaaaa tatccgagaa tattttggaa agtctttgcc agttgatcta
     4201 acgtgtaatc attttggtat tgctcgcata ccaagaacgg acaatgtaga attttttgat
     4261 cctaattacc gttattcttt caaagaatgg caagattggt ctttcaaaca aacagataat
     4321 aagggcttta ctcgttcaag tctaacggtt ttaagcggta cagaaggcaa aaaacaagta
     4381 gatgaaccct ggtttaatct cttattgcac gaaacgaaat tttcaggaga aaagggttta
     4441 atagggcgta ataacgtcat gtttaccctc tctttagcct actttagttc aggctattca
     4501 atcgaaacgt gcgaatataa tatgtttgag tttaataatc gattagatca acccttagaa
     4561 gaaaaagaag taatcaaaat tgttagaagt gcctattcag aaaactatca aggggctaat
     4621 agggaataca ttaccattct ttgcaaagct tgggtatcaa gtgatttaac cagtaaagat
     4681 ttatttgtcc gtcaagggtg gtttaaattc aagaaaaaaa gaagcgaacg tcaacgtgtt
     4741 catttgtcag aatggaaaga agatttaatg gcttatatta gcgaaaaaag cgatgtatac
     4801 aagccttatt tagtgacgac caaaaaagag attagagaag tgctaggcat tcctgaacgg
     4861 acattagata aattgctgaa ggtactgaag gcgaatcagg aaattttctt taagattaaa
     4921 ccaggaagaa atggtggcat tcaacttgct agtgttaaat cattgttgct atcgatcatt
     4981 aaagtaaaaa aagaagaaaa agaaagctat ataaaggcgc tgacaaattc ttttgactta
     5041 gagcatacat tcattcaaga gactttaaac aagctagcag aacgccctaa aacggacaca
     5101 caactcgatt tgtttagcta tgatacaggc tgaaaataaa acccgcacta tgccattaca
     5161 tttatatcta tgatacgtgt ttgttttttc tttgctgttt agcgaatgat tagcagaaat
     5221 atacagagta agattttaat taattattag ggggagaagg agagagtagc ccgaaaactt
     5281 ttagttggct tggactgaac gaagtgaggg aaaggctact aaaacgtcga ggggcagtga
     5341 gagcgaagcg aacacttgat tttttaattt tctatctttt ataggtcatt agagtatact
     5401 tatttgtcct ataaactatt tagcagcata atagatttat tgaataggtc atttaagttg
     5461 agcatattag aggaggaaaa tcttggagaa atatttgaag aacccgatta catggattgg
     5521 attagttctt gtggttacgt ggtttttaac taaaagtagt gaatttttga tttttggtgt
     5581 gtgtgtcttg ttgttagtat ttgctagtca aagtgattaa atagaattcc ggatgagcat
     5641 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
     5701 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     5761 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     5821 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa
     5881 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     5941 gatcaacgtc tcattttcgc caaaagttgg cccagggctt cccggtatca acagggacac
     6001 caggatttat ttattctgcg aagtgatctt ccgtcacagg tatttattcg gcgcaaagtg
     6061 cgtcgggtga tgctgccaac ttactgattt agtgtatgat ggtgtttttg aggtgctcca
     6121 gtggcttctg tttctatcag ctgtccctcc tgttcagcta ctgacggggt ggtgcgtaac
     6181 ggcaaaagca ccgccggaca tcagcgctag cggagtgtat actggcttac tatgttggca
     6241 ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg
     6301 tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact cgctacgctc
     6361 ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga
     6421 tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg
     6481 ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg
     6541 acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt
     6601 cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg
     6661 cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc
     6721 cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag
     6781 acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag ttagtcttga
     6841 agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc gctcctccaa
     6901 gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa ccgccctgca
     6961 aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga tctcaagaag
     7021 atcatcttat taatcagata aaatatttct agatttcagt gcaatttatc tcttcaaatg
     7081 tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgtttg acagcttatc
     7141 atcgataagc tttaatgcgg tagtttatca cagttaaatt gctaacgcag tcaggcaccg
     7201 tgtatgaaat ctaacaatgc gctcatcgtc atcctcggca ccgtcaccct ggatgctgta
     7261 ggcataggct tggttatgcc ggtactgccg ggcctcttgc gggatatcgt ccattccgac
     7321 agcatcgcca gtcactatgg cgtgctgcta gcgctatatg cgttgatgca atttctatgc
     7381 gcacccgttc tcggagcact gtccgaccgc tttggccgcc gcccagtcct gctcgcttcg
     7441 ctacttggag ccactatcga ctacgcgatc atggcgacca cacccgtcct gtggatcctt
     7501 atttgtatag ttcatccatg ccatgtgtaa tcccagcagc cgttacaaac tcaagaagga
     7561 ccatgtggtc tctcttttcg ttgggatctt tcgaaagggc agattgtgtg gacaggtaat
     7621 ggttgtctgg taaaaggaca gggtcatcgc caattggagt attttgttga taatgatcag
     7681 cgagttgcac gccgccgtct tcgatgttgt ggcgggtctt gaagttggct ttgatgccgt
     7741 tcttttgctt gtcggccatg atgtatacgt tgtgggagtt gtagttgtat tccaacttgt
     7801 ggccgaggat gtttccgtcc tccttgaaat cgattccctt aagctcgatc ctgttgacga
     7861 gggtgtctcc ctcaaacttg acttcagcac gtgtcttgta gttcccgtcg tccttgaaga
     7921 agatggtcct ctcctgcacg tatccctcag gcatggcgct cttgaagaag tcgtgccgct
     7981 tcatatgatc tggatatctt gaaaagcatt gaacaccata agtgagagta gtgacaagtg
     8041 ttggccatgg aacaggtagt tttccagtag tgcaaataaa tttaagggta agttttccgt
     8101 atgttgcatc accttcaccc tctccactga cagaaaattt gtacccatta acatcaccat
     8161 ctaattcaac aagaattggg acaactccag tgaaaagttc ttctccttta ctcatgaatt
     8221 caagtctcct tttttattag tgataatttt aacaaagaaa attataccat gttgaagagc
     8281 attaataaaa ttattatttt gtgtttgtgc tattatagtt gagattatta ttaatgaggg
     8341 gtaaataaga tgaagataat tgcaggtttg ggtaatccgg gtcaaaaata tgataagacc
     8401 aaacataata ctggtttcat gacaatggat cactaccttg ataaaaaagg tttgacttta
     8461 aataaagata aatttgaagg gcattggact aaaaagctta tcgataccg
//
