LOCUS       sequence-446948-        3980 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 3980)
  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 3980)
  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-slpGFP
FEATURES             Location/Qualifiers
     source          1..3980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     154..171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    288..309
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(324..329,330..347,348..354)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    362..378
                     /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)."
     primer_bind     367..389
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     386..402
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     386..402
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     421..441
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        423..441
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     470..486
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     471..487
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             804..1520
                     /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(834..862)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1457..1477
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1540..1556)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1540..1556)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1588..1607)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1589..1607)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1617..1634)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1617..1633)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1626..1648)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1775..2230
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1862..1881)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2072..2093
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2256..2360
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2361..2429,2430..3221)
                     /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     complement(2579..2598)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3392..3980
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3881..3900
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      181 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      241 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      301 ctcactcatt aggcacccca ggctttacac tttatgctcc cggctcgtat gttgtgtgga
      361 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
      421 gcaattaacc ctcactaaag ggaacaaaag ctgggtaccg ggccccccct cgaggtcgac
      481 ggtatcgata agcttgatat cgaattcgtg gtaagtaata ggacgtgctt caggcgtgtt
      541 gcctgtacgc atgctgattc ttcagcaaga ctactacctc atgagagtta tagactcatg
      601 gatcttgctt tgaagggttt tgtacattat aggctcctat cacatgctga acctatggcc
      661 tattacattt ttttatattt caaggaggaa aagaccacat gaagaaaaat ttaagaatcg
      721 ttagcgctgc tgctgctgct ttacttgctg ttgctccagt tgctgcttct gctgtatcta
      781 ctgttagcag atccacagcc accatgagta aaggagaaga acttttcact ggagttgtcc
      841 caattcttgt tgaattagat ggtgatgtta atgggtacaa attttctgtc agtggagagg
      901 gtgaaggtga tgcaacatac ggaaaactta cccttaaatt tatttgcact actggaaaac
      961 tacctgttcc atggccaaca cttgtcacta ctctcactta tggtgttcaa tgcttttcaa
     1021 gatatccaga tcatatgaag cggcacgact tcttcaagag cgccatgcct gagggatacg
     1081 tgcaggagag gaccatcttc ttcaaggacg acgggaacta caagacacgt gctgaagtca
     1141 agtttgaggg agacaccctc gtcaacagga tcgagcttaa gggaatcgat ttcaaggagg
     1201 acggaaacat cctcggccac aagttggaat acaactacaa ctcccacaac gtatacatca
     1261 tggccgacaa gcaaaagaac ggcatcaaag ccaacttcaa gacccgccac aacatcgaag
     1321 acggcggcgt gcaactcgct gatcattatc aacaaaatac tccaattggc gatgaccctg
     1381 tccttttacc agacaaccat tacctgtcca cacaatctgc cctttcgaaa gatcccaacg
     1441 aaaagagaga ccacatggtc cttcttgagt ttgtaacggc tgctgggatt acacatggca
     1501 tggatgaact atacaaataa ctcgacctgc agcccggggg atccactagt tctagagcgg
     1561 ccgccaccgc ggtggagctc caattcgccc tatagtgagt cgtattacgc gcgctcactg
     1621 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     1681 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     1741 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     1801 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     1861 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     1921 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     1981 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2041 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2101 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2161 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     2221 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2281 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     2341 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     2401 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2461 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     2521 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     2581 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     2641 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     2701 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     2761 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     2821 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     2881 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     2941 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3001 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3061 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3121 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3181 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     3241 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3301 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3361 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3421 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3481 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     3541 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3601 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3661 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3721 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3781 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3841 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3901 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3961 gggggcggag cctatggaaa
//
