LOCUS       sequence-201585-        8961 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 8961)
  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 8961)
  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-ermGFP
FEATURES             Location/Qualifiers
     source          1..8961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5943..5962
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(6029..6131)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(6523..6540)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6657..7202)
                     /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(6737..6756)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(7314..7319,7320..7336,7337..7342)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             complement(7685..8401)
                     /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(7728..7748)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     8343..8371
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 tcgacctgct gacgaccggc gtcgatattc cgtcgatctg taatatcgtg ttcctgcgta
       61 aagtacgcag ccgcattctg tacgaacaga tgaaaggccg cgccacgcgc ttatgcccgg
      121 aggtgaataa aaccagcttt aagatttttg actgtgtcga tatctacagc acgctggaga
      181 gcgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
      241 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
      301 gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat
      361 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg
      421 tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc
      481 gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat
      541 tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt
      601 agatgacgac catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc
      661 gatcattgga ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg
      721 gttggcatgg attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg
      781 tgcatggagc cgggccacct cgacctgaat ggaagccggc ggcacctcgc taacggattc
      841 accactccaa gaattggagc caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac
      901 ccttggcaga acatatccat cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg
      961 gtcgagtcta gaatcgatac gattttgaag tggcaacaga taaaaaaaag cagtttaaaa
     1021 ttgttgctga acttttaaaa caagcaaata caatcattgt cgcaacagat agcgacagag
     1081 aaggcgaaaa cattgcctgg tcgatcattc ataaagcaaa tgccttttct aaagataaaa
     1141 cgtataaaag actatggatc aatagtttag aaaaagatgt gatccgtagc ggttttcaaa
     1201 atttgcaacc aggaatgaat tactatccct tttatcaaga agcgcacaaa aagaaaaacg
     1261 aaatgataca ccaatcagtg caaaaaaaga tataatggga gataagacgg ttcgtgttcg
     1321 tgctgacttg caccatatca taaaaatcga aacagcaaag aatggcggaa acgtaaaaga
     1381 agttatggaa ataagactta gaagcaaact taagagtgtg ttgatagtgc agtatcttaa
     1441 aattttgtat aataggaatt gaagttaaat tagatgctaa aaatttgtaa ttaagaagga
     1501 gtgattacat gaacaaaaat ataaaatatt ctcaaaactt tttaacgagt gaaaaagtac
     1561 tcaaccaaat aataaaacaa ttgaatttaa aagaaaccga taccgtttac gaaattggaa
     1621 caggtaaagg gcatttaacg acgaaactgg ctaaaataag taaacaggta acgtctattg
     1681 aattagacag tcatctattc aacttatcgt cagaaaaatt aaaactgaat actcgtgtca
     1741 ctttaattca ccaagatatt ctacagtttc aattccctaa caaacagagg tataaaattg
     1801 ttgggagtat tccttaccat ttaagcacac aaattattaa aaaagtggtt tttgaaagcc
     1861 atgcgtctga catctatctg attgttgaag aaggattcta caagcgtacc ttggatattc
     1921 accgaacact agggttgctc ttgcacactc aagtctcgat tcagcaattg cttaagctgc
     1981 cagcggaatg ctttcatcct aaaccaaaag taaacagtgt cttaataaaa cttacccgcc
     2041 ataccacaga tgttccagat aaatattgga agctatatac gtactttgtt tcaaaatggg
     2101 tcaatcgaga atatcgtcaa ctgtttacta aaaatcagtt tcatcaagca atgaaacacg
     2161 ccaaagtaaa caatttaagt accgttactt atgagcaagt attgtctatt tttaatagtt
     2221 atctattatt taacgggagg aaataattct atgagtcgct tttgtaaatt tggaaagtta
     2281 cacgttacta aagggaatgt agataaatta ttaggtatac tactgacagc ttccaaggag
     2341 ctaaagaggt ccctagcgct cttatcatgg ggaagctcgg atcatatgca agacaaaata
     2401 aactcgcaac agcacttgga gaaatgggac gaatcgagaa aaccctcttt acgctggatt
     2461 acatatctaa taaagccgta aggagacggg ttcaaaaagg tttaaataaa ggagaagcaa
     2521 tcaatgcatt agctagaatt atattttttg gacaacgtgg agaatttaga gaacgtgctc
     2581 tccaagacca gttacaaaga gctagtgcac taaacataat tattaacgct ataagtgtgt
     2641 ggaacactgt atatatggaa aaagccgtag aagaattaaa agcaagagga gaatttagag
     2701 aagatttaat gccatatgcg tggccgttag gatgggaaca tatcaatttt cttggagaat
     2761 acaaatttga aggattacat gacactgggc aaatgaattt acgtccttta cgtataaaag
     2821 agccgtttta ttcttaatat aacggctctt tttatagaaa aaatccttag cgtggttttt
     2881 ttccgaaatg ctggcggtac cccaagaatt agaaatgagt agatcaaatt attcacgaat
     2941 agaatcagga aaatcagatc caaccataaa aacactagaa caaattgcaa agttaactaa
     3001 ctcaacgcta gtagtggatt taatcccaaa tgagccaaca gaaccagaac cagaaacaga
     3061 atcagaacaa gtaacattgg atttagaaat ggaagaagaa aaaagcaatg acttcgtgtg
     3121 aataatgcac gaaatcgttg cttatttttt ttaaaagcgg tatactagat ataacgaaac
     3181 aacgaactga atagaaacga aaaaagagcc atgacacatt tataaaatgt ttgacgacat
     3241 tttataaatg catagcccga taagattgcc aaaccaacgc ttatcagtta gtcagatgaa
     3301 ctcttccctc gtaagaagtt atttaattaa ctttgtttga agacggtata taaccgtact
     3361 atcattatat agggaaatca gagagttttc aagtatctaa gctactgaat ttaagaattg
     3421 ttaagcaatc aatcggaaat cgtttgattg ctttttttgt attcatttat agaaggtgga
     3481 gtttgtatga atcatgatga atgtaaaact tatataaaaa atagtttatt ggagataaga
     3541 aaattagcaa atatctatac actagaaacg tttaagaaag agttagaaaa gagaaatatc
     3601 tacttagaaa caaaatcaga taagtatttt tcttcggagg gggaagatta tatatataag
     3661 ttaatagaaa ataacaaaat aatttattcg attagtggaa aaaaattgac ttataaagga
     3721 aaaaaatctt tttcaaaaca tgcaatattg aaacagttga atgaaaaagc aaaccaagtt
     3781 aattaaacaa cctattttat aggatttata ggaaaggaga acagctgaat gaatatccct
     3841 tttgttgtag aaactgtgct tcatgacggc ttgttaaagt acaaatttaa aaatagtaaa
     3901 attcgctcaa tcactaccaa gccaggtaaa agcaaagggg ctatttttgc gtatcgctca
     3961 aaatcaagca tgattggcgg tcgtggtgtt gttctgactt ccgaggaagc gattcaagaa
     4021 aatcaagata catttacaca ttggacaccc aacgtttatc gttatggaac gtatgcagac
     4081 gaaaaccgtt catacacgaa aggacattct gaaaacaatt taagacaaat caataccttc
     4141 tttattgatt ttgatattca cacggcaaaa gaaactattt cagcaagcga tattttaaca
     4201 accgctattg atttaggttt tatgcctact atgattatca aatctgataa aggttatcaa
     4261 gcatattttg ttttagaaac gccagtctat gtgacttcaa aatcagaatt taaatctgtc
     4321 aaagcagcca aaataatttc gcaaaatatc cgagaatatt ttggaaagtc tttgccagtt
     4381 gatctaacgt gtaatcattt tggtattgct cgcataccaa gaacggacaa tgtagaattt
     4441 tttgatccta attaccgtta ttctttcaaa gaatggcaag attggtcttt caaacaaaca
     4501 gataataagg gctttactcg ttcaagtcta acggttttaa gcggtacaga aggcaaaaaa
     4561 caagtagatg aaccctggtt taatctctta ttgcacgaaa cgaaattttc aggagaaaag
     4621 ggtttaatag ggcgtaataa cgtcatgttt accctctctt tagcctactt tagttcaggc
     4681 tattcaatcg aaacgtgcga atataatatg tttgagttta ataatcgatt agatcaaccc
     4741 ttagaagaaa aagaagtaat caaaattgtt agaagtgcct attcagaaaa ctatcaaggg
     4801 gctaataggg aatacattac cattctttgc aaagcttggg tatcaagtga tttaaccagt
     4861 aaagatttat ttgtccgtca agggtggttt aaattcaaga aaaaaagaag cgaacgtcaa
     4921 cgtgttcatt tgtcagaatg gaaagaagat ttaatggctt atattagcga aaaaagcgat
     4981 gtatacaagc cttatttagt gacgaccaaa aaagagatta gagaagtgct aggcattcct
     5041 gaacggacat tagataaatt gctgaaggta ctgaaggcga atcaggaaat tttctttaag
     5101 attaaaccag gaagaaatgg tggcattcaa cttgctagtg ttaaatcatt gttgctatcg
     5161 atcattaaag taaaaaaaga agaaaaagaa agctatataa aggcgctgac aaattctttt
     5221 gacttagagc atacattcat tcaagagact ttaaacaagc tagcagaacg ccctaaaacg
     5281 gacacacaac tcgatttgtt tagctatgat acaggctgaa aataaaaccc gcactatgcc
     5341 attacattta tatctatgat acgtgtttgt tttttctttg ctgtttagcg aatgattagc
     5401 agaaatatac agagtaagat tttaattaat tattaggggg agaaggagag agtagcccga
     5461 aaacttttag ttggcttgga ctgaacgaag tgagggaaag gctactaaaa cgtcgagggg
     5521 cagtgagagc gaagcgaaca cttgattttt taattttcta tcttttatag gtcattagag
     5581 tatacttatt tgtcctataa actatttagc agcataatag atttattgaa taggtcattt
     5641 aagttgagca tattagagga ggaaaatctt ggagaaatat ttgaagaacc cgattacatg
     5701 gattggatta gttcttgtgg ttacgtggtt tttaactaaa agtagtgaat ttttgatttt
     5761 tggtgtgtgt gtcttgttgt tagtatttgc tagtcaaagt gattaaatag aattccggat
     5821 gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt
     5881 tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt
     5941 gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg
     6001 tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata
     6061 actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta
     6121 cgtgccgatc aacgtctcat tttcgccaaa agttggccca gggcttcccg gtatcaacag
     6181 ggacaccagg atttatttat tctgcgaagt gatcttccgt cacaggtatt tattcggcgc
     6241 aaagtgcgtc gggtgatgct gccaacttac tgatttagtg tatgatggtg tttttgaggt
     6301 gctccagtgg cttctgtttc tatcagctgt ccctcctgtt cagctactga cggggtggtg
     6361 cgtaacggca aaagcaccgc cggacatcag cgctagcgga gtgtatactg gcttactatg
     6421 ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc
     6481 ggtgcgtcag cagaatatgt gatacaggat atattccgct tcctcgctca ctgactcgct
     6541 acgctcggtc gttcgactgc ggcgagcgga aatggcttac gaacggggcg gagatttcct
     6601 ggaagatgcc aggaagatac ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc
     6661 cataggctcc gcccccctga caagcatcac gaaatctgac gctcaaatca gtggtggcga
     6721 aacccgacag gactataaag ataccaggcg tttccccctg gcggctccct cgtgcgctct
     6781 cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg tttgtctcat
     6841 tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact gtatgcacga
     6901 accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6961 ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta gaggagttag
     7021 tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt gactgcgctc
     7081 ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc
     7141 cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca aaacgatctc
     7201 aagaagatca tcttattaat cagataaaat atttctagat ttcagtgcaa tttatctctt
     7261 caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca tgtttgacag
     7321 cttatcatcg ataagcttta atgcggtagt ttatcacagt taaattgcta acgcagtcag
     7381 gcaccgtgta tgaaatctaa caatgcgctc atcgtcatcc tcggcaccgt caccctggat
     7441 gctgtaggca taggcttggt tatgccggta ctgccgggcc tcttgcggga tatcgtccat
     7501 tccgacagca tcgccagtca ctatggcgtg ctgctagcgc tatatgcgtt gatgcaattt
     7561 ctatgcgcac ccgttctcgg agcactgtcc gaccgctttg gccgccgccc agtcctgctc
     7621 gcttcgctac ttggagccac tatcgactac gcgatcatgg cgaccacacc cgtcctgtgg
     7681 atccttattt gtatagttca tccatgccat gtgtaatccc agcagccgtt acaaactcaa
     7741 gaaggaccat gtggtctctc ttttcgttgg gatctttcga aagggcagat tgtgtggaca
     7801 ggtaatggtt gtctggtaaa aggacagggt catcgccaat tggagtattt tgttgataat
     7861 gatcagcgag ttgcacgccg ccgtcttcga tgttgtggcg ggtcttgaag ttggctttga
     7921 tgccgttctt ttgcttgtcg gccatgatgt atacgttgtg ggagttgtag ttgtattcca
     7981 acttgtggcc gaggatgttt ccgtcctcct tgaaatcgat tcccttaagc tcgatcctgt
     8041 tgacgagggt gtctccctca aacttgactt cagcacgtgt cttgtagttc ccgtcgtcct
     8101 tgaagaagat ggtcctctcc tgcacgtatc cctcaggcat ggcgctcttg aagaagtcgt
     8161 gccgcttcat atgatctgga tatcttgaaa agcattgaac accataagtg agagtagtga
     8221 caagtgttgg ccatggaaca ggtagttttc cagtagtgca aataaattta agggtaagtt
     8281 ttccgtatgt tgcatcacct tcaccctctc cactgacaga aaatttgtac ccattaacat
     8341 caccatctaa ttcaacaaga attgggacaa ctccagtgaa aagttcttct cctttactca
     8401 tgaattcact ccttcttaat tacaaatttt tagcatctaa tttaacttca attcctatta
     8461 tacaaaattt taagatactg cactatcaac acactcttaa gtttgcttct aagtcttatt
     8521 tccataactt cttttacgtt tccgccattc tttgctgttt cgatttttat gatatggtgc
     8581 aagtcagcac gaacacgaac cgtcttatct cccattatat ctttttttgc actgattggt
     8641 gtatcatttc gtttttcttt ttgtgcgctt cttgataaaa gggatagtaa ttcattcctg
     8701 gttgcaaatt ttgaaaaccg ctacggatca catctttttc taaactattg atccatagtc
     8761 ttttatacgt tttatcttta gaaaaggcat ttgctttatg aatgatcgac caggcaatgt
     8821 tttcgccttc tctgtcgcta tctgttgcga caatgattgt atttgcttgt tttaaaagtt
     8881 cagcaacaat tttaaactgc ttttttttat ctgttgccac ttcaaaatcg tatcgattct
     8941 agactaagct tatcgatacc g
//
