LOCUS       sequence-377516-        5426 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 5426)
  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 5426)
  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-135
FEATURES             Location/Qualifiers
     source          1..5426
                     /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             1776..2471
                     /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(1909..1927)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2177..2196
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      2492..2586
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2630..3289
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2696..2715)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3354..3440
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3816..3838
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3830..3847
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3831..3847
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     oriT            3935..4043
                     /note="incP origin of transfer"
     protein_bind    4190..4211
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4226..4231,4232..4249,4250..4256)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4264..4280
                     /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     4269..4291
                     /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 ttcgagctcg cacgctgccg caagcactca
     1441 gggcgcaagg gctgctaaag gaagcggaac acgtagaaag ccagtccgca gaaacggtgc
     1501 tgaccccgga tgaatgtcag ctactgggct atctggacaa gggaaaacgc aagcgcaaag
     1561 agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc ggtcttatgg
     1621 acagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc
     1681 aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagatct
     1741 gatcaagaga caggatacta gtggaggaag aaaaaatggt gagcaagggc gaggcagtga
     1801 tcaaggagtt catgcggttc aaggtgcaca tggagggctc catgaacggc cacgagttcg
     1861 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     1921 tgaccaaggg tggccccctg cccttctcct gggacatcct gtcccctcag ttcatgtacg
     1981 gctccagggc cttcatcaag caccccgccg acatccccga ctactataag cagtccttcc
     2041 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgcc gtgaccgtga
     2101 cccaggacac ctccctggag gacggcaccc tgatctacaa ggtgaagctc cgcggcacca
     2161 acttccctcc tgacggcccc gtaatgcaga agaagacaat gggctgggaa gcgtccaccg
     2221 agcggttgta ccccgaggac ggcgtgctga agggcgacat taagatggcc ctgcgcctga
     2281 aggacggcgg ccgctacctg gcggacttca agaccaccta caaggccaag aagcccgtgc
     2341 agatgcccgg cgcctacaac gtcgaccgca agttggacat cacctcccac aacgaggact
     2401 acaccgtggt ggaacagtac gaacgctccg agggccgcca ctccaccggc ggcatggacg
     2461 agctgtacaa gtaagtaatt agctgagctt ggactcctgt tgatagatcc agtaatgacc
     2521 tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
     2581 gagaatccaa gctagcttgg cgagattttc aggagctaag gaagctaaaa tggagaaaaa
     2641 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
     2701 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
     2761 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
     2821 ccgcctgatg aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat
     2881 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
     2941 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
     3001 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     3061 cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     3121 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     3181 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
     3241 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg
     3301 cagttattgg tgcccttaaa cgcctggggt aatgactctc tagcttgagg catcaaataa
     3361 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     3421 ctctcctgag taggacaaat ccgccgctag gagcttgcgg ccgccagccc gggcaattcg
     3481 aggggtatcg ctcttgaagg gaactatgtt gaataataaa aacgaatttt cacaaaaaaa
     3541 tgtgtttttc tttgttagac cggggactta tcagccaacc tgttatgtgg cgcggtatta
     3601 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3661 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3721 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3781 atcgaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     3841 ggccagtgaa ttaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     3901 aatgtcatga taataatggt ttcttagagc ttatcggcca gcctcgcaga gcaggattcc
     3961 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg
     4021 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac
     4081 gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg
     4141 ctataatgac cccgaagcag ggttatgcag cgggtgagcg caacgcaatt aatgtgagtt
     4201 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     4261 gaattgtgag cggataacaa tttcacacag gaaacacata tggaaaaaaa ggaatttcgt
     4321 gttttgataa aatactgttt tctgaaggga aaaaatacag tggaagcaaa aacttggctt
     4381 gataatgagt ttccggactc tgccccaggg aaatcaacaa taattgattg gtatgcaaaa
     4441 ttcaagcgtg gtgaaatgag cacggaggac ggtgaacgca gtggacgccc gaaagaggtg
     4501 gttaccgacg aaaacatcaa aaaaatccac aaaatgattt tgaatgaccg taaaatgaag
     4561 ttgatcgaga tagcagaggc cttaaagata tcaaaggaac gtgttggtca tatcattcat
     4621 caatatttgg atatgcggaa gctctgtgca aaatgggtgc cgcgcgagct cacatttgac
     4681 caaaaacaac aacgtgttga tgattctgag cggtgtttgc agctgttaac tcgtaataca
     4741 cccgagtttt tccgtcgata tgtgacaatg gatgaaacat ggctccatca ctacactcct
     4801 gagtccaatc gacagtcggc tgagtggaca gcgaccggtg aaccgtctcc gaagcgtgga
     4861 aagactcaaa agtccgctgg caaagtaatg gcctctgttt tttgggatgc gcatggaata
     4921 atttttatcg attatcttga gaagggaaaa accatcaaca gtgactatta tatggcgtta
     4981 ttggagcgtt tgaaggtcga aatcgcggca aaacggcccc acatgaagaa gaaaaaagtg
     5041 ttgttccacc aagacaacgc accgtgccac aagtcattga gaacgatggc aaaaattcat
     5101 gaattgggct tcgaattgct tccccacccg ccgtattctc cagatctggc ccccagcgac
     5161 tttttcttgt tctcagacct caaaaggatg ctcgcaggga aaaaatttgg ctgcaatgaa
     5221 gaggtgatcg ccgaaactga ggcctatttt gaggcaaaac cgaaggagta ctaccaaaat
     5281 ggtatcaaaa aattggaagg tcgttataat cgttgtatcg ctcttgaagg gaactatgtt
     5341 gaataataaa aacgaatttt cacaaaaaaa tgtgtttttc tttgttagac cggatctgca
     5401 gaattccagc acactggcgg ccgtta
//
