LOCUS       sequence-11333-e        4198 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 4198)
  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 4198)
  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: pBS-ermGFP
FEATURES             Location/Qualifiers
     source          1..4198
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(8..24)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9..25)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(54..74)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(54..72)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(93..109)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(93..109)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(106..128)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    117..133
                     /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(141..147,148..165,166..171))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    186..207
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(324..341)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(495..1083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(575..594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1254..2045,2046..2114))
                     /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     1877..1896
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2115..2219)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2245..2700)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2382..2403)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2594..2613
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2827..2849
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2841..2858
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2842..2858
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2868..2887
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2868..2886
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2919..2935
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2919..2935
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             complement(2937..3653)
                     /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(2980..3000)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     3595..3623
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 agcttatcga taccgtcgac ctcgaggggg ggcccggtac ccagcttttg ttccctttag
       61 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
      121 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
      181 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
      241 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
      301 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
      361 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
      421 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      481 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
      541 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
      601 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
      661 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
      721 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
      781 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
      841 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
      901 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
      961 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1021 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1081 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     1141 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     1201 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     1261 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     1321 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     1381 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     1441 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     1501 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     1561 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     1621 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     1681 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     1741 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     1801 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     1861 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     1921 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     1981 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     2041 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     2101 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     2161 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     2221 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     2281 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     2341 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     2401 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     2461 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     2521 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     2581 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     2641 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     2701 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     2761 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     2821 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac
     2881 tatagggcga attggagctc caccgcggtg gcggccgctc tagaactagt ggatccttat
     2941 ttgtatagtt catccatgcc atgtgtaatc ccagcagccg ttacaaactc aagaaggacc
     3001 atgtggtctc tcttttcgtt gggatctttc gaaagggcag attgtgtgga caggtaatgg
     3061 ttgtctggta aaaggacagg gtcatcgcca attggagtat tttgttgata atgatcagcg
     3121 agttgcacgc cgccgtcttc gatgttgtgg cgggtcttga agttggcttt gatgccgttc
     3181 ttttgcttgt cggccatgat gtatacgttg tgggagttgt agttgtattc caacttgtgg
     3241 ccgaggatgt ttccgtcctc cttgaaatcg attcccttaa gctcgatcct gttgacgagg
     3301 gtgtctccct caaacttgac ttcagcacgt gtcttgtagt tcccgtcgtc cttgaagaag
     3361 atggtcctct cctgcacgta tccctcaggc atggcgctct tgaagaagtc gtgccgcttc
     3421 atatgatctg gatatcttga aaagcattga acaccataag tgagagtagt gacaagtgtt
     3481 ggccatggaa caggtagttt tccagtagtg caaataaatt taagggtaag ttttccgtat
     3541 gttgcatcac cttcaccctc tccactgaca gaaaatttgt acccattaac atcaccatct
     3601 aattcaacaa gaattgggac aactccagtg aaaagttctt ctcctttact catgaattca
     3661 ctccttctta attacaaatt tttagcatct aatttaactt caattcctat tatacaaaat
     3721 tttaagatac tgcactatca acacactctt aagtttgctt ctaagtctta tttccataac
     3781 ttcttttacg tttccgccat tctttgctgt ttcgattttt atgatatggt gcaagtcagc
     3841 acgaacacga accgtcttat ctcccattat atcttttttt gcactgattg gtgtatcatt
     3901 tcgtttttct ttttgtgcgc ttcttgataa aagggatagt aattcattcc tggttgcaaa
     3961 ttttgaaaac cgctacggat cacatctttt tctaaactat tgatccatag tcttttatac
     4021 gttttatctt tagaaaaggc atttgcttta tgaatgatcg accaggcaat gttttcgcct
     4081 tctctgtcgc tatctgttgc gacaatgatt gtatttgctt gttttaaaag ttcagcaaca
     4141 attttaaact gctttttttt atctgttgcc acttcaaaat cgtatcgatt ctagacta
//
