LOCUS       sequence-383671-        6280 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Himar transposon delivery plasmid that delivers a red fluorescent 
            mScarlet-I protein gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6280)
  AUTHORS   Stocks C, Schlechter R, Remus-Emsermann M
  TITLE     Chromatic Bacteria v.2-A Himar1 Transposon-Based Delivery Vector to 
            Extend the Host Range of a Toolbox to Fluorescently Tag Bacteria
  JOURNAL   Bacteria 2022, 1(1), 56-65
REFERENCE   2  (bases 1 to 6280)
  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: "Bacteria 
            2022, 1(1), 56-65"
COMMENT     Sequence Label: pMRE-Himar-155
FEATURES             Location/Qualifiers
     source          1..6280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      588..976
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             1474..2268
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     GQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDVGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             2630..3325
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(2763..2781)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3031..3050
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      3346..3440
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3484..4143
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3550..3569)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      4208..4294
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4670..4692
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4684..4701
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4685..4701
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     oriT            4789..4897
                     /note="incP origin of transfer"
     protein_bind    5044..5065
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5080..5085,5086..5103,5104..5110)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5118..5134
                     /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     5123..5145
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 ctagatgcat gctcgagcgg ccgccagtgt gatggattga aaaaggaaga gtatgagtat
       61 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
      121 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
      181 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
      241 ttttccaatg atgagcactt ttaaagttct gctaacaggt tggctgataa gtccccggtc
      301 tgacacatag atggcgtcgc tagtattaaa tgcatattat ttttatatag taccaacctt
      361 caaatgattc ccagatctca aactggaaca acactcaacc ctatctcggt ctattctttt
      421 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
      481 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
      541 gaaatgtgcg cggaaccaga tccggccacg atgcgtccgg cgtagaggat ctgaagatca
      601 gcagttcaac ctgttgatag tacgtactaa gctctcatgt ttcacgtact aagctctcat
      661 gtttaacgta ctaagctctc atgtttaacg aactaaaccc tcatggctaa cgtactaagc
      721 tctcatggct aacgtactaa gctctcatgt ttcacgtact aagctctcat gtttgaacaa
      781 taaaattaat ataaatcagc aacttaaata gcctctaagg ttttaagttt tataagaaaa
      841 aaaagaatat ataaggcttt taaagctttt aaggtttaac ggttgtggac aacaagccag
      901 ggatgtaacg cactgagaag cccttagagc ctctcaaagc aattttgagt gacacaggaa
      961 cacttaacgg ctgacatggg aattcttgaa gacgaaaggg cctcgtgata cgcctatttt
     1021 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggtt
     1081 aattaagggc tgcaggaatt cgatatcaag cttatcgata ccgtcgaaat tctgattaat
     1141 aatttgttta aatttttcgt ttggcgtgag gtatccaagt cttttacgag gtcgattatt
     1201 gagtttattt tcaatccact taatctgttt gttggttact tcactaaagt ccttaccctt
     1261 tgggatatac tgcctgataa gcccgttggt gttttcattg gcaccacgtt cccatgagtg
     1321 gtatggtttg caaaaataga attttatttc caatttttgc gcaatttcct cgtgctctag
     1381 aatataaact gccaggaatt ggggatcgga ttcgagctaa gaattccctt ggggtatctt
     1441 taaatactgt agaaaagagg aaggaaataa taaatggcta aaatgagaat atcaccggaa
     1501 ttgaaaaaac tgatcgaaaa ataccgctgc gtaaaagata cggaaggaat gtctcctgct
     1561 aaggtatata agctggtggg agaaaatgaa aacctatatt taaaaatgac ggacagccgg
     1621 tataaaggga ccacctatga tgtggaacgg gaaaaggaca tgatgctatg gctggaagga
     1681 aagctgcctg ttccaaaggt cctgcacttt gaacggcatg atggctggag caatctgctc
     1741 atgagtgagg ccgatggcgt cctttgctcg gaagagtatg aagatggaca aagccctgaa
     1801 aagattatcg agctgtatgc ggagtgcatc aggctctttc actccatcga catatcggat
     1861 tgtccctata cgaatagctt agacagccgc ttagccgaat tggattactt actgaataac
     1921 gatctggccg atgtggattg cgaaaactgg gaagaagaca ctccatttaa agatccgcgc
     1981 gagctgtatg attttttaaa gacggaaaag cccgaagagg aacttgtctt ttcccacggc
     2041 gacctgggag acagcaacat ctttgtgaaa gatggcaaag taagtggctt tattgatctt
     2101 gggagaagcg gcagggcgga caagtggtat gacattgcct tctgcgtccg gtcgatcagg
     2161 gaggatgtcg gggaagaaca gtatgtcgag ctattttttg acttactggg gatcaagcct
     2221 gattgggaga aaataaaata ttatatttta ctggatgaat tgttttagag ctcgcacgct
     2281 gccgcaagca ctcagggcgc aagggctgct aaaggaagcg gaacacgtag aaagccagtc
     2341 cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg acaagggaaa
     2401 acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga tagctagact
     2461 gggcggtctt atggacagca agcgaaccgg aattgccagc tggggcgccc tctggtaagg
     2521 ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca
     2581 ggggatcaag atctgatcaa gagacaggat actagtggag gaagaaaaaa tggtgagcaa
     2641 gggcgaggca gtgatcaagg agttcatgcg gttcaaggtg cacatggagg gctccatgaa
     2701 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     2761 cgccaagctg aaggtgacca agggtggccc cctgcccttc tcctgggaca tcctgtcccc
     2821 tcagttcatg tacggctcca gggccttcat caagcacccc gccgacatcc ccgactacta
     2881 taagcagtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     2941 cgccgtgacc gtgacccagg acacctccct ggaggacggc accctgatct acaaggtgaa
     3001 gctccgcggc accaacttcc ctcctgacgg ccccgtaatg cagaagaaga caatgggctg
     3061 ggaagcgtcc accgagcggt tgtaccccga ggacggcgtg ctgaagggcg acattaagat
     3121 ggccctgcgc ctgaaggacg gcggccgcta cctggcggac ttcaagacca cctacaaggc
     3181 caagaagccc gtgcagatgc ccggagccta caacgtcgac cgcaagttgg acatcacctc
     3241 ccacaacgag gactacaccg tggtggaaca gtacgaacgc tccgagggcc gccactccac
     3301 cggcggcatg gacgagctgt acaagtaagt aattagctga gcttggactc ctgttgatag
     3361 atccagtaat gacctcagaa ctccatctgg atttgttcag aacgctcggt tgccgccggg
     3421 cgttttttat tggtgagaat ccaagctagc ttggcgagat tttcaggagc taaggaagct
     3481 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa
     3541 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg
     3601 gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt
     3661 attcacattc ttgcccgcct gatgaatgct catccggaat ttcgtatggc aatgaaagac
     3721 ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact
     3781 gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata
     3841 tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt
     3901 gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac
     3961 gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa
     4021 ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc
     4081 catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg
     4141 taattttttt aaggcagtta ttggtgccct taaacgcctg gggtaatgac tctctagctt
     4201 gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     4261 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ctaggagctt gcggccgcca
     4321 gcccgggcaa ttcgaggggt atcgctcttg aagggaacta tgttgaataa taaaaacgaa
     4381 ttttcacaaa aaaatgtgtt tttctttgtt agaccgggga cttatcagcc aacctgttat
     4441 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4501 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4561 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4621 tacttctgac aacgatcgag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     4681 cgttgtaaaa cgacggccag tgaattaatt cttgaagacg aaagggcctc gtgatacgcc
     4741 tatttttata ggttaatgtc atgataataa tggtttctta gagcttatcg gccagcctcg
     4801 cagagcagga ttcccgttga gcaccgccag gtgcgaataa gggacagtga agaaggaaca
     4861 cccgctcgcg ggtgggccta cttcacctat cctgcccggc tgacgccgtt ggatacacca
     4921 aggaaagtct acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata
     4981 taccgaaaaa atcgctataa tgaccccgaa gcagggttat gcagcgggtg agcgcaacgc
     5041 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     5101 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca catatggaaa
     5161 aaaaggaatt tcgtgttttg ataaaatact gttttctgaa gggaaaaaat acagtggaag
     5221 caaaaacttg gcttgataat gagtttccgg actctgcccc agggaaatca acaataattg
     5281 attggtatgc aaaattcaag cgtggtgaaa tgagcacgga ggacggtgaa cgcagtggac
     5341 gcccgaaaga ggtggttacc gacgaaaaca tcaaaaaaat ccacaaaatg attttgaatg
     5401 accgtaaaat gaagttgatc gagatagcag aggccttaaa gatatcaaag gaacgtgttg
     5461 gtcatatcat tcatcaatat ttggatatgc ggaagctctg tgcaaaatgg gtgccgcgcg
     5521 agctcacatt tgaccaaaaa caacaacgtg ttgatgattc tgagcggtgt ttgcagctgt
     5581 taactcgtaa tacacccgag tttttccgtc gatatgtgac aatggatgaa acatggctcc
     5641 atcactacac tcctgagtcc aatcgacagt cggctgagtg gacagcgacc ggtgaaccgt
     5701 ctccgaagcg tggaaagact caaaagtccg ctggcaaagt aatggcctct gttttttggg
     5761 atgcgcatgg aataattttt atcgattatc ttgagaaggg aaaaaccatc aacagtgact
     5821 attatatggc gttattggag cgtttgaagg tcgaaatcgc ggcaaaacgg ccccacatga
     5881 agaagaaaaa agtgttgttc caccaagaca acgcaccgtg ccacaagtca ttgagaacga
     5941 tggcaaaaat tcatgaattg ggcttcgaat tgcttcccca cccgccgtat tctccagatc
     6001 tggcccccag cgactttttc ttgttctcag acctcaaaag gatgctcgca gggaaaaaat
     6061 ttggctgcaa tgaagaggtg atcgccgaaa ctgaggccta ttttgaggca aaaccgaagg
     6121 agtactacca aaatggtatc aaaaaattgg aaggtcgtta taatcgttgt atcgctcttg
     6181 aagggaacta tgttgaataa taaaaacgaa ttttcacaaa aaaatgtgtt tttctttgtt
     6241 agaccggatc tgcagaattc cagcacactg gcggccgtta
//
