LOCUS       sequence-451369-        9087 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Replicative vector for orange fluorescent reporter protein mKO2 
            strong constitutive recombinant expression in C. acnes. pBRESP36A 
            with P(BBa_J23119)+RBS_1+mKO2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9087)
  AUTHORS   Nevot G, Santos-Moreno J, Campama-Sanz N, Toloza L, Parra-Cid C, 
            Jansen PAM, Barbier I, Ledesma-Amaro R, van den Bogaard EH, Guell M
  TITLE     Synthetically programmed antioxidant delivery by a domesticated skin
            commensal.
  JOURNAL   Cell Syst. 2025 Feb 19;16(2):101169. doi: 
            10.1016/j.cels.2025.101169. Epub 2025 Feb 6.
  PUBMED    39919749
REFERENCE   2  (bases 1 to 9087)
  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: "Cell 
            Syst."; date: "2025-02-19"; volume: "16(2):101169. doi"; pages: " 
            10.1016/j.cels.2025.101169. Epub 2025 Feb 6"
COMMENT     Sequence Label: pEND-225_Cacnes_mKO2
FEATURES             Location/Qualifiers
     source          1..9087
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4120..4311
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4305..4327)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4413..4553
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4468..4487
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4568..4585)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4739..5327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4819..4838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5498..6289,6290..6358))
                     /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     6121..6140
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6359..6463)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6531..6549
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    6614..6632
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     misc_feature    8000..8018
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     promoter        8271..8305
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             8395..9048
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     CDS             9049..9066
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 ctcgagcaat aaacagttga tagggcttct ccgttaccat ggttcagcca aaaaacttaa
       61 gaccgccggt cttgtccact accttgcagt aatgcggtgg acaggatcgg cggttttctt
      121 ttctcttctc aagctagcga aatccctgaa actgagactg tagaaaataa gcttccgaaa
      181 ctgaacctgt tattttacgc tacggacgcc cggtcatgac gctgagttag tggcgcagct
      241 gcgctctgag aacgagtttt tacggcgtca ggtcgagcag caggcgcgca cgatcgaacg
      301 gcaggctgag gcacacgcgg tggtctcagc gcagctcaca cgggttggcc agcttgaggc
      361 cggcgacgca gcagcaccga cactggcacc cgttgaaagg ccggctccgc gacggcggtg
      421 gtggcagcgt cggtagcggt caggatcgct ctggcgtgac gagtgtgtct ggcagtgcga
      481 acagttgctc gaccagtggc agcagaagcg agatcgctgc gtggtgctgt tcctcggtca
      541 gttcgtcgag gactggcggg tcttgctgcg tccagccgat cgcctcggcg gccaaggtca
      601 gttccaagct gtgccaacgc acacgcccct cggctgacag ctgagtctcg aactgtgcaa
      661 ctggaccggc cggaagatgc acgttgccga ggtcgtgagt ggccaagcgc acgtcaaaga
      721 gtgctgcttc gtagccgcgc agaaatggca gtgctcggtc gattcggatc ggcctgccca
      781 ggtacattcc gggccgcttg atgaacgcct ccgcgtagaa gcgcaccgtt ctcggcccgg
      841 cctcgtgatc tgtcactgtg cacgctcctc tcgatggttc tcgacgctac cggagaccac
      901 cgacgttcat gcccagcgca gcgacctgaa aggaccaagc cgagttagcc gtgctaaccg
      961 tatagcttgc tccgtcgcct ctgagggcaa ccacctgcgc agcaggtggg cggcagcccg
     1021 cgcgcaagcg cctaccgggt ttgggcacag cccataaatc aacgcctccg gtgttgaagc
     1081 gatcgtgtgt cacgattgct atgcttgcta ccccttcagg gttttcgtat acacaaatca
     1141 agttttttcg tatacgctaa tgccatgagt gagcatctac tgcacggcaa gcccgtcacc
     1201 aacgagcaga ttcaggcatg ggcagacgag gccgaggccg gatacgacct gcccaaactc
     1261 cccaagccac ggcgcggacg cccgcccgta ggagacggtc cgggcaccgt cgtacccgtg
     1321 cgtctcgacg cggccaccgt tgccgctctc acagaacgag caacagccga gggcatcacg
     1381 aaccgttcag acgcgatccg agccgcagtc cacgagtgga cacgggttgc ctgacctcca
     1441 cgactcagca cgcaagcact accaacgaga ccggctcgac gacacggccg tgctctacgc
     1501 ggccacccac gttctcaact cccggccact cgacgacgaa gacgacccgc gccgctggct
     1561 catgatcgga accgacccag caggccgcct actcgaactc gtcgcactga tctacgacga
     1621 cggctacgaa ctgatcatcc acgcaatgaa agcccgcacc caatacctcg accagctcta
     1681 accaagaaag gaacctgatg agcgaccagc tagacagcga ccgcaactac gacccgatga
     1741 tcttcgacgt gatgcgcgag accgcgaacc gcgtcgtcgc cacgtacgtt gcatgggaag
     1801 atgaagccgc tgatccccgc gaggctgcgc actggcaggc cgagcgattc cgcacccggc
     1861 acgaggtgcg cgccgtcgac cctgacagcc ggcgagcagt tcaggcgaag atcgcacagc
     1921 tgcgcgagga actagccgca atgcccgaac acgccccagc catcccttgg agcaggtggc
     1981 agcgtcaggg gagtcggggg atgtttggca ggggatgtgg aaagagagtt cgctttgctc
     2041 acatggctca accgggtaac taactgatat ggggtcttcg tcgcccactt tgaacacgcc
     2101 gaggaatgga ccacgctgaa cgtgactcgc atgcttcact gcatgtatgg attcgttcga
     2161 gacgttgttc cctgagagct ggctgccacg caagccgctg gcgtcagccg agaagtctgg
     2221 ggcgtaccgg cacgtgactc ggcagagggc gctggagctg ccttacatcg aagcgaaccc
     2281 gttggtcatg cagtccttgg tcatcaccga tcgagatgct tcggatgctg actgggccgc
     2341 agacctcgct gggctgcctt caccgtccta cgtgtccatg aaccgtgtca cgaccaccgg
     2401 acacatcgtc tatgccttga agaaccctgt gtgtctgacc gatgccgcgc ggcgacggcc
     2461 tatcaacctg ctcgcccgcg tcgagcaggg cctatgcgac gttctcggcg gcgatgcatc
     2521 ctacgggcac cggatcacaa agaacccgct cagcaccgcc catgcgaccc tctggggccc
     2581 cgcagacgcg ctctacgagc tgcgcgccct cgcacacacc ctcgacgaga tccacgcact
     2641 gccggaggca gggaacccgc gtcgcaacgt cacccgatca acggtcggcc gcaacgtcac
     2701 cctgttcgac accacccgca tgtgggcata ccgggccgtc cggcactcct ggggcggccc
     2761 ggtcgccgaa tgggagcaca ccgtattcga gcacatccac ctactgaacg agacgatcat
     2821 cgccgacgaa ttcgccacag gccccctcgg cttgaacgaa cttaagcact tatctcgatc
     2881 catttcccga tgggtctggc gcaacttcac ccccgaaacc ttccgcgcac gccagaaagc
     2941 gatcagcctc cgtggagcat ccaaaggcgg caaagaaggc ggccacaaag gcggcattgc
     3001 cagtggcgca tcacggcgcg cccatacccg tcaacagttc ttggagggtc tctcatgacc
     3061 acacgtgaac gtctcccccg caacggctac agcatcgccg ctgctgcgaa aaagctcggt
     3121 gtctccgagt ccaccgtcaa gcggtggact tccgagccac gcgaggagtt cgtggcccgc
     3181 gttgccgcac gccacgcgcg gattcgtgag ctgcgctcgg agggtcagag catgcgtgcg
     3241 attgctgccg aggtcggggt ttccgtgggc accgtgcact acgcgctgaa caagaatcga
     3301 actgacgcat gaccgtaacg ccgcacgatg agcattttct tgatcgtgca ccgcttggca
     3361 ctacgttcgc gtgcggttgc acagtgcgcg ccacgttctt atcctgcggc cattgtggct
     3421 acagccaatg gggggcatca gcaacggacg ttgaacccgg tgggcaagtg ttactcaggg
     3481 ggacatgccc agtctgcggc gctcggattg acggtatggc agtcgtgcat gcggccccac
     3541 cgtcaaactc attcaggtat cagtgagaac cctcatggca ccccctcgtg acacgttctc
     3601 gttgcgatca gctgcggcca agcttgatct tcgggcagcg ttgggtcctg gccacgggtg
     3661 cgcatgatcg tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt
     3721 tagcagaatg aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct
     3781 gcgacctgag caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg
     3841 cggaagtcag cgccctgcac cattatgttc cggatctgca tcgcaggatg ctgctggcta
     3901 ccctgtggaa cacctacatc tgtattaacg aagcgctggc attgaccctg agtgattttt
     3961 ctctggtccc gccgcatcca taccgccagt tgtttaccct cacaacgttc cagtaaccgg
     4021 gcatgttcat catcagtaac ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt
     4081 acccccatga acagaaatcc cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc
     4141 gcccttaaca tggcccgctt tatcagaagc cagacattaa cgcttctgga gaaactcaac
     4201 gagctggacg cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag
     4261 ctttaccgca gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     4321 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     4381 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
     4441 agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     4501 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
     4561 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4621 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4681 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4741 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4801 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4861 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4921 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4981 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5041 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5101 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5161 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5221 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5281 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5341 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5401 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5461 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5521 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5581 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5641 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5701 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5761 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca
     5821 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5881 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5941 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6001 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6061 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg
     6121 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6181 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6241 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6301 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6361 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6421 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6481 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     6541 tcacgaggcc ctttcgtctt caagaattca agcttgtcga cgttaacctg caggcatgcg
     6601 gatccggtac cagctgtctc ttatacacat ctctgcagtt aacgctgacc cggaccacca
     6661 cgacgacccg gacgaccacc ggtacgacca ccaccagacg gaccgcggcg accaccctgg
     6721 tcaccctgtt cacgttcacc ggtacgaccg tcgtcacggt tacgaccaga agaacgacgg
     6781 tcaccgtgac cagacgcttg ccggtctctg cgatcttggc ccccggtacg tctgtcacca
     6841 cggtcctgac caccggtacg acggtcgccc ctatcctgtc cacccgtcct acgaccacgc
     6901 tgcggctgac caccacgcgg ttcgtttttc tggtccagac gttcgaacag acgcagccac
     6961 tgttccggac gaacgtaagc aacaacagca ccaccaccaa caccagcgtg gtccagagca
     7021 gcttcaacac gacgacgcgg gtatttacgc agcagagaag cctggatgtt accaccaaca
     7081 ccggtgaagc acagttcaac catagattcg taacgttcca gagcagcacc ttccagcagc
     7141 ggttcagcac gacgacgcag acgcatgata gcagagtcaa ctttcggaac cggtttgaac
     7201 agacgacgat cgactttttc aacgaattcc cattcgaaca gcggccaggt cataacggtc
     7261 agacgagacc aacgaccgta gtcaccggta cgtttacgag cgaattccag ctgggtaacc
     7321 atggtagcgg tttcgatggt cggagcttcc aggcaccagt caacgatagc agaggtgata
     7381 ccgtacggga tagcaccaac gaaagcgaac ggttccggcg gcggttcagc ggtcaggaag
     7441 tcagcgttaa caacttcgat gttcgggtga ccagacagtt tttcacgcag agatttagcc
     7501 agacgcgggt cgatttcgta agaggtaacc tgacgagcac ggtcagccag ttcacgggtc
     7561 agcagacctt cacccggacc agcttccaga accggcaggt ccggacgcag ttcagcggtt
     7621 tcagcgatac gagcgatggt tttacggtca cgcaggaagt tctggccgaa ttggcggcgg
     7681 ttacggtcac gctcggtacg gccgaggacg gggcgattct ggttggggtg ctggcggtct
     7741 tggttgcggc ggcgggggcg gggctgttca tcggaggagg acatgacgat ctcccgatgt
     7801 gtcgtacgta tgggaattga cgaaaatcat atagattttc agttgaagtg tagcactgca
     7861 tagtgtcatc gaagtgtcgg cggcaacggt atgcccggta gggggtgggt ctgcctttcg
     7921 gtttcgtgaa cctgatgcgg cgacgcgcat gtagccgata acccctagac tcgcccgtgc
     7981 agatcgtcgg tgtggatcca gatgtgtata agagacagct ggtaccggcc gctgctgtgc
     8041 gtgcgggcgt gagcgtttct acgctgcggc gcaggaaatc agagcttgag gctgccggag
     8101 cgacggtaga cccgtccggt tgggtggtgc cactgcgtgc actcaaggtc gtttttgggg
     8161 tgtcagatga gacctcgaat gcgaatacca gtaaatcctg acctgtgagt tgtgaattca
     8221 taggatagat tctggaaact ttaccgtccg agctccagcc tgcggtccgg ttgacagcta
     8281 gctcagtcct aggtataatg ctagctcgct gggacgcccg gtagtgctta tcagacccaa
     8341 tactgttgaa cagcctgcgg tccggtccac ttataaatat agaggaggtt tgatatggtt
     8401 tcggttatca aaccagagat gaaaatgcgt tactatatgg atggttcagt aaatggtcac
     8461 gaatttacta ttgagggcga gggtacggga cgcccatacg aggggcacca ggaaatgact
     8521 ttacgcgtca caatggctga aggcgggcct atgccgtttg cgttcgatct tgttagtcat
     8581 gtcttttgtt acggtcaccg tgtatttact aaataccccg aggaaattcc agactatttc
     8641 aaacaagcct tcccggaagg tttgtcttgg gagcgcagtt tagagtttga agacggtggc
     8701 tcggccagcg tgtcagctca tattagtctt cgcggcaata cattttatca caagtcaaag
     8761 ttcaccggcg tgaacttccc cgcagacggc ccaatcatgc agaatcaaag tgttgattgg
     8821 gaaccgtcca cagagaagat tacagcttcc gatggagtct taaagggcga tgtaaccatg
     8881 tacttaaaat tagaaggggg agggaaccat aaatgtcaga tgaagactac ctataaggcc
     8941 gcaaaagaga ttcttgaaat gcccggagac cactacattg ggcatcgttt ggtccgtaag
     9001 acagaaggaa atattactga acaggtcgaa gacgctgtgg cacacagcca tcatcaccat
     9061 caccactgat aatcgctggg acgcccg
//
