LOCUS       sequence-380552-        5411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  rpsM promoter driving the expression of destabilized GFPmut3 
            (containing a LVA C-terminal tail).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5411)
  AUTHORS   Avraham, Roi
  TITLE     Bacterial fluorescent reporters and tags toolkit
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5411)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pFPV25.1_dGFPmut3-LVA
FEATURES             Location/Qualifiers
     source          1..5411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(96..684)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(176..195)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(855..1646,1647..1715))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1478..1497
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1716..1820)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1888..1906
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             2574..3287
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             2574..3287
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2604..2632)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3227..3247
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     oriT            3936..4023
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     primer_bind     complement(5336..5353)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
       61 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
      121 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
      181 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
      241 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
      301 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
      361 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
      421 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
      481 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
      541 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
      601 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
      661 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
      721 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
      781 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
      841 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
      901 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
      961 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     1021 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     1081 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     1141 aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt
     1201 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     1261 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     1321 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     1381 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     1441 cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga
     1501 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     1561 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     1621 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     1681 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     1741 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     1801 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     1861 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa
     1921 gaattcgagc tcggtacccg gggatcccgg agttcatgcg tgatgcaatg aaagtgccgt
     1981 tctacttcgg tgggacctca ctgcttatcg ttgttgtcgt gattatggac tttatggctc
     2041 aagtgcaaac tctgatgatg tccagtcagt atgagtctgc attgaagaag gcgaacctga
     2101 aaggctacgg ccgttaattg gtcgcctgag aagttacgga gagtaaaaat gaaagttcgt
     2161 gcttccgtca agaaattatg ccgtaactgc aaaatcgtta agcgtgatgg tgtcatccgt
     2221 gtgatttgca gtgccgagcc gaagcataaa cagcgccaag gctgattttt tcgcatattt
     2281 ttcttgcaaa gttgggttga gctggctaga ttagccagcc aatcttttgt atgtctgtac
     2341 gtttccattt gagtatcctg aaaacgggct tttcagcatg gtacgtacat attaaatagt
     2401 aggagtgcat agtggcccgt atagcaggca ttaacattcc tgatcagaaa cacgccgtga
     2461 tcgcgttaac ttcgatctac ggtgtcggca agacccgttc taaagccatc ctggctgcag
     2521 cgggtatcgc tgaaaatgtt aagatcctct agatttaaga aggagatata catatgagta
     2581 aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta
     2641 atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac ggaaaactta
     2701 cccttaaatt tatttgcact actggaaaac tacctgttcc atggccaaca cttgtcacta
     2761 ctttcggtta tggtgttcaa tgctttgcga gatacccaga tcatatgaaa cagcatgact
     2821 ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg
     2881 acgggaacta caagacacgt gctgaagtca agtttgaagg tgataccctt gttaatagaa
     2941 tcgagttaaa aggtattgat tttaaagaag atggaaacat tcttggacac aaattggaat
     3001 acaactataa ctcacacaat gtatacatca tggcagacaa acaaaagaat ggaatcaaag
     3061 ttaacttcaa aattagacac aacattgaag atggaagcgt tcaactagca gaccattatc
     3121 aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat tacctgtcca
     3181 cacaatctgc cctttcgaaa gatcccaacg aaaagagaga ccacatggtc cttcttgagt
     3241 ttgtaacagc tgctgggatt acacatggca tggatgaact atacaaaagg cctgcagcaa
     3301 acgacgaaaa ctacgcttta gtagcttaaa gctttaatgc ggtagtttat cacagttaaa
     3361 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
     3421 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
     3481 gcgggatatc gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctttt
     3541 agccgcttta gcggcctttc cccctacccg aagggtgggg gcgcgtgtgc agccccgcag
     3601 ggcctgtctc ggtcgatcat tcagcccggc tcatccttct ggcgtggcgg cagaccgaac
     3661 aaggcgcggt cgtggtcgcg ttcaaggtac gcatccattg ccgccatgag ccgatcctcc
     3721 ggccactcgc tgctgttcac cttggccaaa atcatggccc ccaccagcac cttgcgcctt
     3781 gtttcgttct tgcgctcttg ctgctgttcc cttgcccgca cccgctgaat ttcggcattg
     3841 attcgcgctc gttgttcttc gagcttggcc agccgatccg ccgccttgtt gctcccctta
     3901 accatcttga caccccattg ttaatgtgct gtctcgtagg ctatcatgga ggcacagcgg
     3961 cggcaatccc gaccctactt tgtaggggag ggggcactta ccggtttctc ttcgagaaac
     4021 tggcctaacg gccacccttc gggcggtgcg ctctccgagg gccattgcat ggagccgaaa
     4081 agcaaaagca acagcgaggc agcatggcga tttatcacct tacggcgaaa accggcagca
     4141 ggtcgggcgg ccaatcggcc agggccaagg ccgactacat ccagcgcgaa ggcaagtatg
     4201 cccgcgacat ggatgaagtc ttgcacgccg aatccgggca catgccggag ttcgtcgagc
     4261 ggcccgccga ctactgggat gctgccgacc tgtatgaacg cgccaatggg cggctgttca
     4321 aggaggtcga atttgccctg ccggtcgagc tgaccctcga ccagcagaag gcgctggcgt
     4381 ccgagttcgc ccagcacctg accggtgccg agcgcctgcc gtatacgctg gccatccatg
     4441 ccggtggcgg cgagaacccg cactgccacc tgatgatctc cgagcggatc aatgacggca
     4501 tcgagcggcc cgccgctcag tggttcaagc ggtacaacgg caagaccccg gagaagggcg
     4561 gggcacagaa gaccgaagcg ctcaagccca aggcatggct tgagcagacc cgcgaggcat
     4621 gggccgacca tgccaaccgg gcattagagc gggctggcca cgacgcccgc attgaccaca
     4681 gaacacttga ggcgcagggc atcgagcgcc tgcccggtgt tcacctgggg ccgaacgtgg
     4741 tggagatgga aggccggggc atccgcaccg accgggcaga cgtggccctg aacatcgaca
     4801 ccgccaacgc ccagatcatc gacttacagg aataccggga ggcaatagac catgaacgca
     4861 atcgacagag tgaagaaatc cagaggcatc aacgagttag cggagcagat cgaaccgctg
     4921 gcccagagca tggcgacact ggccgacgaa gcccggcagg tcatgagcca gacccagcag
     4981 gccagcgagg cgcaggcggc ggagtggctg aaagcccagc gccagacagg ggcggcatgg
     5041 gtggagctgg ccaaagagtt gcgggaggta gccgccgagg tgagcagcgc cgcgcagagc
     5101 gcccggagcg cgtcgcgggg gtggcactgg aagctatggc taaccgtgat gctggcttcc
     5161 atgatgccta cggtggtgct gctgatcgca tcgttgctct tgctcgacct gacgccactg
     5221 acaaccgagg acggctcgat ctggctgcgc ttggtggccc gatgaagaac gacaggactt
     5281 tgcaggccat aggccgacag ctcaaggcca tgggctgtga gcgctcttcc gcttcctcgc
     5341 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5401 cggtaatacg g
//
