LOCUS       sequence-449603-        7540 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses periplasmic mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7540)
  AUTHORS   Heesterbeek DAC, Martin NI, Velthuizen A, Duijst M, Ruyken M, 
            Wubbolts R, Rooijakkers SHM, Bardoel BW
  TITLE     Complement-dependent outer membrane perturbation sensitizes 
            Gram-negative bacteria to Gram-positive specific antibiotics.
  JOURNAL   Sci Rep. 2019 Feb 28;9(1):3074. doi: 10.1038/s41598-019-38577-9.
  PUBMED    30816122
REFERENCE   2  (bases 1 to 7540)
  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: "Sci Rep."; 
            date: "2019-02-28"; volume: "9(1):3074. doi"; pages: " 
            10.1038/s41598-019-38577-9"
COMMENT     Sequence Label: pPerimCh
FEATURES             Location/Qualifiers
     source          1..7540
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            871..958
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             complement(1437..2150)
                     /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"
     primer_bind     complement(1477..1497)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     2092..2120
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     promoter        complement(2228..2512)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(2286..2305)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2539..3417
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     CDS             complement(3679..4386)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(3957..3976)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4226..4244
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     sig_peptide     complement(4393..4458)
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     primer_bind     complement(5117..5135)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5203..5307
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5308..5376,5377..6168)
                     /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     complement(5526..5545)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6339..6927
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6828..6847
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7081..7098
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gttgatgcct ctggatttct tcactctgtc gattgcgttc atggtctatt gcctcccggt
       61 attcctgtaa gtcgatgatc tgggcgttgg cggtgtcgat gttcagggcc acgtctgccc
      121 ggtcggtgcg gatgccccgg ccttccatct ccaccacgtt cggccccagg tgaacaccgg
      181 gcaggcgctc gatgccctgc gcctcaagtg ttctgtggtc aatgcgggcg tcgtggccag
      241 cccgctctaa tgcccggttg gcatggtcgg cccatgcctc gcgggtctgc tcaagccatg
      301 ccttgggctt gagcgcttcg gtcttctgtg ccccgccctt ctccggggtc ttgccgttgt
      361 accgcttgaa ccactgagcg gcgggccgct cgatgccgtc attgatccgc tcggagatca
      421 tcaggtggca gtgcgggttc tcgccgccac cggcatggat ggccagcgta tacggcaggc
      481 gctcggcacc ggtcaggtgc tgggcgaact cggacgccag cgccttctgc tggtcgaggg
      541 tcagctcgac cggcagggca aattcgacct ccttgaacag ccgcccattg gcgcgttcat
      601 acaggtcggc agcatcccag tagtcggcgg gccgctcgac gaactccggc atgtgcccgg
      661 attcggcgtg caagacttca tccatgtcgc gggcatactt gccttcgcgc tggatgtagt
      721 cggccttggc cctggccgat tggccgcccg acctgctgcc ggttttcgcc gtaaggtgat
      781 aaatcgccat gctgcctcgc tgttgctttt gcttttcggc tccatgcaat ggccctcgga
      841 gagcgcaccg cccgaagggt ggccgttagg ccagtttctc gaagagaaac cggtaagtgc
      901 gccctcccct acaaagtagg gtcgggattg ccgccgctgt gcctccatga tagcctacga
      961 gacagcacat taacaatggg gtgtcaagat ggttaagggg agcaacaagg cggcggatcg
     1021 gctggccaag ctcgaagaac aacgagcgcg aatcaatgcc gaaattcagc gggtgcgggc
     1081 aagggaacag cagcaagagc gcaagaacga aacaaggcgc aaggtgctgg tgggggccat
     1141 gattttggcc aaggtgaaca gcagcgagtg gccggaggat cggctcatgg cggcaatgga
     1201 tgcgtacctt gaacgcgacc acgaccgcgc cttgttcggt ctgccgccac gccagaagga
     1261 tgagccgggc tgaatgatcg accgagacag gccctgcggg gctgcacacg cgcccccacc
     1321 cttcgggtag ggggaaaggc cgctaaagcg gctaaaagcg ctagcagcac gccatagtga
     1381 ctggcgatgc tgtcggaatg gacgatagct tgcatgcctg caggtctgga catttatttg
     1441 tatagttcat ccatgccatg tgtaatccca gcagctgtta caaactcaag aaggaccatg
     1501 tggtctctct tttcgttggg atctttcgaa agggcagatt gtgtggacag gtaatggttg
     1561 tctggtaaaa ggacagggcc atcgccaatt ggagtatttt gttgataatg gtctgctagt
     1621 tgaacgcttc catcttcaat gttgtgtcta attttgaagt taactttgat tccattcttt
     1681 tgtttgtctg ccatgatgta tacattgtgt gagttatagt tgtattccaa tttgtgtcca
     1741 agaatgtttc catcttcttt aaaatcaata ccttttaact cgattctatt aacaagggta
     1801 tcaccttcaa acttgacttc agcacgtgtc ttgtagttcc cgtcatcttt gaaaaatata
     1861 gttctttcct gtacataacc ttcgggcatg gcactcttga aaaagtcatg ctgtttcata
     1921 tgatctgggt atctcgcaaa gcattgaaca ccataaccga aagtagtgac aagtgttggc
     1981 catggaacag gtagttttcc agtagtgcaa ataaatttaa gggtaagttt tccgtatgtt
     2041 gcatcacctt caccctctcc actgacagaa aatttgtgcc cattaacatc accatctaat
     2101 tcaacaagaa ttgggacaac tccagtgaaa agttcttctc ctttactcat atgtatatct
     2161 ccttcttaaa tctagaggat ccccgggtac catggtgaat tcctcctgct agcccaaaaa
     2221 aacgggtatg gagaaacagt agagagttgc gataaaaagc gtcaggtagg atccgctaat
     2281 cttatggata aaaatgctat ggcatagcaa agtgtgacgc cgtgcaaata atcaatgtgg
     2341 acttttctgc cgtgattata gacacttttg ttacgcgttt ttgtcatggc tttggtcccg
     2401 ctttgttaca gaatgctttt aataagcggg gttaccggtt tggttagcga gaagagccag
     2461 taaaagacgc agtgacggca atgtctgatg caatatggac aattggtttc ttctctgaat
     2521 ggcgggagta tgaaaagtat ggctgaagcg caaaatgatc ccctgctgcc gggatactcg
     2581 tttaatgccc atctggtggc gggtttaacg ccgattgagg ccaacggtta tctcgatttt
     2641 tttatcgacc gaccgctggg aatgaaaggt tatattctca atctcaccat tcgcggtcag
     2701 ggggtggtga aaaatcaggg acgagaattt gtttgccgac cgggtgatat tttgctgttc
     2761 ccgccaggag agattcatca ctacggtcgt catccggagg ctcgcgaatg gtatcaccag
     2821 tgggtttact ttcgtccgcg cgcctactgg catgaatggc ttaactggcc gtcaatattt
     2881 gccaatacgg ggttctttcg cccggatgaa gcgcaccagc cgcatttcag cgacctgttt
     2941 gggcaaatca ttaacgccgg gcaaggggaa gggcgctatt cggagctgct ggcgataaat
     3001 ctgcttgagc aattgttact gcggcgcatg gaagcgatta acgagtcgct ccatccaccg
     3061 atggataatc gggtacgcga ggcttgtcag tacatcagcg atcacctggc agacagcaat
     3121 tttgatatcg ccagcgtcgc acagcatgtt tgcttgtcgc cgtcgcgtct gtcacatctt
     3181 ttccgccagc agttagggat tagcgtctta agctggcgcg aggaccaacg tatcagccag
     3241 gcgaagctgc ttttgagcac cacccggatg cctatcgcca ccgtcggtcg caatgttggt
     3301 tttgacgatc aactctattt ctcgcgggta tttaaaaaat gcaccggggc cagcccgagc
     3361 gagttccgtg ccggttgtga agaaaaagtg aatgatgtag ccgtcaagtt gtcataattg
     3421 gtaacgaatc agacaattga cggcttgacg gagtagcata gggtttgcag aatccctgct
     3481 tcgtccattt gacaggcaca ttatgcatcg atcccgcaag aggcccggca gtaccggcat
     3541 aaccaagcct atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt
     3601 gttagatttc atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca
     3661 ttaaagcttt cgcggccgct acttgtacag ctcgtccatg ccgccggtgg agtggcggcc
     3721 ctcggcgcgt tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg tgatgtccaa
     3781 cttgatgttg acgttgtagg cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt
     3841 cttgacctca gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct tgatctcgcc
     3901 cttcagggcg ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc ccatggtctt
     3961 cttctgcatt acggggccgt cggaggggaa gttggtgccg cgcagcttca ccttgtagat
     4021 gaactcgccg tcctgcaggg aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa
     4081 gttcatcacg cgctcccact tgaagccctc ggggaaggac agcttcaagt agtcggggat
     4141 gtcggcgggg tgcttcacgt aggccttgga gccgtacatg aactgagggg acaggatgtc
     4201 ccaggcgaag ggcagggggc cacccttggt caccttcagc ttggcggtct gggtgccctc
     4261 gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg tggccgttca cggagccctc
     4321 catgtgcacc ttgaagcgca tgaactcctt gatgatggcc atgttatcct cctcgccctt
     4381 gctcacagat ctagccatgg ccggctgggc ggccagcagc agcaggccag ctgcagcggt
     4441 cggcagcagg tacttcatct cgagttctcc tcccaggatc ttaacatttt cagcgatacc
     4501 cgctgcagcc aggatggctt tagaacgggt cttgccgaca ccgtagatcg aagttaacgc
     4561 gatcacggcg tgtttctgat caggaatgtt aatgcctgct atacgggcca ctatgcactc
     4621 ctactattta atatgtacgt accatgctga aaagcccgtt ttcaggatac tcaaatggaa
     4681 acgtacagac atacaaaaga ttggctggct aatctagcca gctcaaccca actttgcaag
     4741 aaaaatatgc gaaaaaatca gccttggcgc tgtttatgct tcggctcggc actgcaaatc
     4801 acacggatga caccatcacg cttaacgatt ttgcagttac ggcataattt cttgacggaa
     4861 gcacgaactt tcatttttac tctccgtaac ttctcaggcg accaattaac ggccgtagcc
     4921 tttcaggttc gccttcttca atgcagactc atactgactg gacatcatca gagtttgcac
     4981 ttgagccata aagtccataa tcacgacaac aacgataagc agtgaggtcc caccgaagta
     5041 gaacggcact ttcattgcat cacgcatgaa ctccgggatg cccgggttcc gagctcgaat
     5101 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     5161 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5221 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5281 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5341 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5401 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5461 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5521 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     5581 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5641 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5701 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5761 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5821 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     5881 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5941 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6001 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6061 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6121 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6181 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6241 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6301 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6361 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6421 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6481 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6541 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6601 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6661 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6721 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6781 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6841 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6901 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6961 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     7021 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     7081 agcgagtcag tgagcgagga agcggaagag cgctcacagc ccatggcctt gagctgtcgg
     7141 cctatggcct gcaaagtcct gtcgttcttc atcgggccac caagcgcagc cagatcgagc
     7201 cgtcctcggt tgtcagtggc gtcaggtcga gcaagagcaa cgatgcgatc agcagcacca
     7261 ccgtaggcat catggaagcc agcatcacgg ttagccatag cttccagtgc cacccccgcg
     7321 acgcgctccg ggcgctctgc gcggcgctgc tcacctcggc ggctacctcc cgcaactctt
     7381 tggccagctc cacccatgcc gcccctgtct ggcgctgggc tttcagccac tccgccgcct
     7441 gcgcctcgct ggcctgctgg gtctggctca tgacctgccg ggcttcgtcg gccagtgtcg
     7501 ccatgctctg ggccagcggt tcgatctgct ccgctaactc
//
