LOCUS       sequence-352653-        8560 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 8560)
  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 8560)
  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-slpGFP
FEATURES             Location/Qualifiers
     source          1..8560
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5753..5772
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5839..5941)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(6333..6350)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6467..7012)
                     /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(6547..6566)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(7124..7129,7130..7146,7147..7152)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             complement(7513..8229)
                     /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(7556..7576)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     8171..8199
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc
       61 gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg
      121 ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg
      181 cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc
      241 aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac
      301 gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct
      361 tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac
      421 catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc gatcattgga
      481 ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg gttggcatgg
      541 attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc
      601 cgggccacct cgacctgaat ggaagccggc ggcacctcgc taacggattc accactccaa
      661 gaattggagc caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac ccttggcaga
      721 acatatccat cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg gtcgagtcta
      781 gaatcgatac gattttgaag tggcaacaga taaaaaaaag cagtttaaaa ttgttgctga
      841 acttttaaaa caagcaaata caatcattgt cgcaacagat agcgacagag aaggcgaaaa
      901 cattgcctgg tcgatcattc ataaagcaaa tgccttttct aaagataaaa cgtataaaag
      961 actatggatc aatagtttag aaaaagatgt gatccgtagc ggttttcaaa atttgcaacc
     1021 aggaatgaat tactatccct tttatcaaga agcgcacaaa aagaaaaacg aaatgataca
     1081 ccaatcagtg caaaaaaaga tataatggga gataagacgg ttcgtgttcg tgctgacttg
     1141 caccatatca taaaaatcga aacagcaaag aatggcggaa acgtaaaaga agttatggaa
     1201 ataagactta gaagcaaact taagagtgtg ttgatagtgc agtatcttaa aattttgtat
     1261 aataggaatt gaagttaaat tagatgctaa aaatttgtaa ttaagaagga gtgattacat
     1321 gaacaaaaat ataaaatatt ctcaaaactt tttaacgagt gaaaaagtac tcaaccaaat
     1381 aataaaacaa ttgaatttaa aagaaaccga taccgtttac gaaattggaa caggtaaagg
     1441 gcatttaacg acgaaactgg ctaaaataag taaacaggta acgtctattg aattagacag
     1501 tcatctattc aacttatcgt cagaaaaatt aaaactgaat actcgtgtca ctttaattca
     1561 ccaagatatt ctacagtttc aattccctaa caaacagagg tataaaattg ttgggagtat
     1621 tccttaccat ttaagcacac aaattattaa aaaagtggtt tttgaaagcc atgcgtctga
     1681 catctatctg attgttgaag aaggattcta caagcgtacc ttggatattc accgaacact
     1741 agggttgctc ttgcacactc aagtctcgat tcagcaattg cttaagctgc cagcggaatg
     1801 ctttcatcct aaaccaaaag taaacagtgt cttaataaaa cttacccgcc ataccacaga
     1861 tgttccagat aaatattgga agctatatac gtactttgtt tcaaaatggg tcaatcgaga
     1921 atatcgtcaa ctgtttacta aaaatcagtt tcatcaagca atgaaacacg ccaaagtaaa
     1981 caatttaagt accgttactt atgagcaagt attgtctatt tttaatagtt atctattatt
     2041 taacgggagg aaataattct atgagtcgct tttgtaaatt tggaaagtta cacgttacta
     2101 aagggaatgt agataaatta ttaggtatac tactgacagc ttccaaggag ctaaagaggt
     2161 ccctagcgct cttatcatgg ggaagctcgg atcatatgca agacaaaata aactcgcaac
     2221 agcacttgga gaaatgggac gaatcgagaa aaccctcttt acgctggatt acatatctaa
     2281 taaagccgta aggagacggg ttcaaaaagg tttaaataaa ggagaagcaa tcaatgcatt
     2341 agctagaatt atattttttg gacaacgtgg agaatttaga gaacgtgctc tccaagacca
     2401 gttacaaaga gctagtgcac taaacataat tattaacgct ataagtgtgt ggaacactgt
     2461 atatatggaa aaagccgtag aagaattaaa agcaagagga gaatttagag aagatttaat
     2521 gccatatgcg tggccgttag gatgggaaca tatcaatttt cttggagaat acaaatttga
     2581 aggattacat gacactgggc aaatgaattt acgtccttta cgtataaaag agccgtttta
     2641 ttcttaatat aacggctctt tttatagaaa aaatccttag cgtggttttt ttccgaaatg
     2701 ctggcggtac cccaagaatt agaaatgagt agatcaaatt attcacgaat agaatcagga
     2761 aaatcagatc caaccataaa aacactagaa caaattgcaa agttaactaa ctcaacgcta
     2821 gtagtggatt taatcccaaa tgagccaaca gaaccagaac cagaaacaga atcagaacaa
     2881 gtaacattgg atttagaaat ggaagaagaa aaaagcaatg acttcgtgtg aataatgcac
     2941 gaaatcgttg cttatttttt ttaaaagcgg tatactagat ataacgaaac aacgaactga
     3001 atagaaacga aaaaagagcc atgacacatt tataaaatgt ttgacgacat tttataaatg
     3061 catagcccga taagattgcc aaaccaacgc ttatcagtta gtcagatgaa ctcttccctc
     3121 gtaagaagtt atttaattaa ctttgtttga agacggtata taaccgtact atcattatat
     3181 agggaaatca gagagttttc aagtatctaa gctactgaat ttaagaattg ttaagcaatc
     3241 aatcggaaat cgtttgattg ctttttttgt attcatttat agaaggtgga gtttgtatga
     3301 atcatgatga atgtaaaact tatataaaaa atagtttatt ggagataaga aaattagcaa
     3361 atatctatac actagaaacg tttaagaaag agttagaaaa gagaaatatc tacttagaaa
     3421 caaaatcaga taagtatttt tcttcggagg gggaagatta tatatataag ttaatagaaa
     3481 ataacaaaat aatttattcg attagtggaa aaaaattgac ttataaagga aaaaaatctt
     3541 tttcaaaaca tgcaatattg aaacagttga atgaaaaagc aaaccaagtt aattaaacaa
     3601 cctattttat aggatttata ggaaaggaga acagctgaat gaatatccct tttgttgtag
     3661 aaactgtgct tcatgacggc ttgttaaagt acaaatttaa aaatagtaaa attcgctcaa
     3721 tcactaccaa gccaggtaaa agcaaagggg ctatttttgc gtatcgctca aaatcaagca
     3781 tgattggcgg tcgtggtgtt gttctgactt ccgaggaagc gattcaagaa aatcaagata
     3841 catttacaca ttggacaccc aacgtttatc gttatggaac gtatgcagac gaaaaccgtt
     3901 catacacgaa aggacattct gaaaacaatt taagacaaat caataccttc tttattgatt
     3961 ttgatattca cacggcaaaa gaaactattt cagcaagcga tattttaaca accgctattg
     4021 atttaggttt tatgcctact atgattatca aatctgataa aggttatcaa gcatattttg
     4081 ttttagaaac gccagtctat gtgacttcaa aatcagaatt taaatctgtc aaagcagcca
     4141 aaataatttc gcaaaatatc cgagaatatt ttggaaagtc tttgccagtt gatctaacgt
     4201 gtaatcattt tggtattgct cgcataccaa gaacggacaa tgtagaattt tttgatccta
     4261 attaccgtta ttctttcaaa gaatggcaag attggtcttt caaacaaaca gataataagg
     4321 gctttactcg ttcaagtcta acggttttaa gcggtacaga aggcaaaaaa caagtagatg
     4381 aaccctggtt taatctctta ttgcacgaaa cgaaattttc aggagaaaag ggtttaatag
     4441 ggcgtaataa cgtcatgttt accctctctt tagcctactt tagttcaggc tattcaatcg
     4501 aaacgtgcga atataatatg tttgagttta ataatcgatt agatcaaccc ttagaagaaa
     4561 aagaagtaat caaaattgtt agaagtgcct attcagaaaa ctatcaaggg gctaataggg
     4621 aatacattac cattctttgc aaagcttggg tatcaagtga tttaaccagt aaagatttat
     4681 ttgtccgtca agggtggttt aaattcaaga aaaaaagaag cgaacgtcaa cgtgttcatt
     4741 tgtcagaatg gaaagaagat ttaatggctt atattagcga aaaaagcgat gtatacaagc
     4801 cttatttagt gacgaccaaa aaagagatta gagaagtgct aggcattcct gaacggacat
     4861 tagataaatt gctgaaggta ctgaaggcga atcaggaaat tttctttaag attaaaccag
     4921 gaagaaatgg tggcattcaa cttgctagtg ttaaatcatt gttgctatcg atcattaaag
     4981 taaaaaaaga agaaaaagaa agctatataa aggcgctgac aaattctttt gacttagagc
     5041 atacattcat tcaagagact ttaaacaagc tagcagaacg ccctaaaacg gacacacaac
     5101 tcgatttgtt tagctatgat acaggctgaa aataaaaccc gcactatgcc attacattta
     5161 tatctatgat acgtgtttgt tttttctttg ctgtttagcg aatgattagc agaaatatac
     5221 agagtaagat tttaattaat tattaggggg agaaggagag agtagcccga aaacttttag
     5281 ttggcttgga ctgaacgaag tgagggaaag gctactaaaa cgtcgagggg cagtgagagc
     5341 gaagcgaaca cttgattttt taattttcta tcttttatag gtcattagag tatacttatt
     5401 tgtcctataa actatttagc agcataatag atttattgaa taggtcattt aagttgagca
     5461 tattagagga ggaaaatctt ggagaaatat ttgaagaacc cgattacatg gattggatta
     5521 gttcttgtgg ttacgtggtt tttaactaaa agtagtgaat ttttgatttt tggtgtgtgt
     5581 gtcttgttgt tagtatttgc tagtcaaagt gattaaatag aattccggat gagcattcat
     5641 caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt
     5701 ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga
     5761 ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc
     5821 agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa
     5881 tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc
     5941 aacgtctcat tttcgccaaa agttggccca gggcttcccg gtatcaacag ggacaccagg
     6001 atttatttat tctgcgaagt gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc
     6061 gggtgatgct gccaacttac tgatttagtg tatgatggtg tttttgaggt gctccagtgg
     6121 cttctgtttc tatcagctgt ccctcctgtt cagctactga cggggtggtg cgtaacggca
     6181 aaagcaccgc cggacatcag cgctagcgga gtgtatactg gcttactatg ttggcactga
     6241 tgagggtgtc agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag
     6301 cagaatatgt gatacaggat atattccgct tcctcgctca ctgactcgct acgctcggtc
     6361 gttcgactgc ggcgagcgga aatggcttac gaacggggcg gagatttcct ggaagatgcc
     6421 aggaagatac ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc cataggctcc
     6481 gcccccctga caagcatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag
     6541 gactataaag ataccaggcg tttccccctg gcggctccct cgtgcgctct cctgttcctg
     6601 cctttcggtt taccggtgtc attccgctgt tatggccgcg tttgtctcat tccacgcctg
     6661 acactcagtt ccgggtaggc agttcgctcc aagctggact gtatgcacga accccccgtt
     6721 cagtccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggaaagacat
     6781 gcaaaagcac cactggcagc agccactggt aattgattta gaggagttag tcttgaagtc
     6841 atgcgccggt taaggctaaa ctgaaaggac aagttttggt gactgcgctc ctccaagcca
     6901 gttacctcgg ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc
     6961 ggttttttcg ttttcagagc aagagattac gcgcagacca aaacgatctc aagaagatca
     7021 tcttattaat cagataaaat atttctagat ttcagtgcaa tttatctctt caaatgtagc
     7081 acctgaagtc agccccatac gatataagtt gtaattctca tgtttgacag cttatcatcg
     7141 ataagcttta atgcggtagt ttatcacagt taaattgcta acgcagtcag gcaccgtgta
     7201 tgaaatctaa caatgcgctc atcgtcatcc tcggcaccgt caccctggat gctgtaggca
     7261 taggcttggt tatgccggta ctgccgggcc tcttgcggga tatcgtccat tccgacagca
     7321 tcgccagtca ctatggcgtg ctgctagcgc tatatgcgtt gatgcaattt ctatgcgcac
     7381 ccgttctcgg agcactgtcc gaccgctttg gccgccgccc agtcctgctc gcttcgctac
     7441 ttggagccac tatcgactac gcgatcatgg cgaccacacc cgtcctgtgg atcccccggg
     7501 ctgcaggtcg agttatttgt atagttcatc catgccatgt gtaatcccag cagccgttac
     7561 aaactcaaga aggaccatgt ggtctctctt ttcgttggga tctttcgaaa gggcagattg
     7621 tgtggacagg taatggttgt ctggtaaaag gacagggtca tcgccaattg gagtattttg
     7681 ttgataatga tcagcgagtt gcacgccgcc gtcttcgatg ttgtggcggg tcttgaagtt
     7741 ggctttgatg ccgttctttt gcttgtcggc catgatgtat acgttgtggg agttgtagtt
     7801 gtattccaac ttgtggccga ggatgtttcc gtcctccttg aaatcgattc ccttaagctc
     7861 gatcctgttg acgagggtgt ctccctcaaa cttgacttca gcacgtgtct tgtagttccc
     7921 gtcgtccttg aagaagatgg tcctctcctg cacgtatccc tcaggcatgg cgctcttgaa
     7981 gaagtcgtgc cgcttcatat gatctggata tcttgaaaag cattgaacac cataagtgag
     8041 agtagtgaca agtgttggcc atggaacagg tagttttcca gtagtgcaaa taaatttaag
     8101 ggtaagtttt ccgtatgttg catcaccttc accctctcca ctgacagaaa atttgtaccc
     8161 attaacatca ccatctaatt caacaagaat tgggacaact ccagtgaaaa gttcttctcc
     8221 tttactcatg gtggctgtgg atctgctaac agtagataca gcagaagcag caactggagc
     8281 aacagcaagt aaagcagcag cagcagcgct aacgattctt aaatttttct tcatgtggtc
     8341 ttttcctcct tgaaatataa aaaaatgtaa taggccatag gttcagcatg tgataggagc
     8401 ctataatgta caaaaccctt caaagcaaga tccatgagtc tataactctc atgaggtagt
     8461 agtcttgctg aagaatcagc atgcgtacag gcaacacgcc tgaagcacgt cctattactt
     8521 accacgaatt cgatatcaag cttatcgata ccgtcgaccg
//
