LOCUS       sequence-377514-        7132 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 7132)
  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 7132)
  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-175
FEATURES             Location/Qualifiers
     source          1..7132
                     /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             3482..4177
                     /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(3615..3633)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3883..3902
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      4198..4292
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             4336..4995
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4402..4421)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      5060..5146
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5522..5544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5536..5553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5537..5553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     oriT            5641..5749
                     /note="incP origin of transfer"
     protein_bind    5896..5917
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5932..5937,5938..5955,5956..5962)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5970..5986
                     /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     5975..5997
                     /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 cgtgctttgc
     1381 aaactccttt ccattgtcag ccgtaattgt gtgtattaag tttttcactt tccgcagtgc
     1441 ccatactgca atcttagcta ccgggatggc ttcttttccc gacaacttgc gtatccagac
     1501 cctgcttgtt gctctgtcgt taatggtaag aatggcacct ttgtggttct taccaataat
     1561 tgtatctatc tctaaatcac caaatctctc cttcagttcc actatctcgg gacgctcatc
     1621 aatatccacc ctgcctggga taaatcctcg ccctgcattt ttagaaccac gtttggcata
     1681 cctgcgacct tgtctgcgaa gatatttgtg cagtttgcca ccccgccgct tatcctccca
     1741 aatccagcga tatatcgttt cgtgagatac catcgcaatt ccctccaagc ggctcctgcc
     1801 gacaatctgc tccgggctga atcctttctt caacagcttt attatccgtt ttctcattgc
     1861 cggtgtaagc acttccttgc gatgtttttg ctgcttgcgc ctgtctgctt ttcgctgggc
     1921 aagctccatg ctatagctac cacttcgggc gtcgcaattg cgctttatct ccctgtaaac
     1981 agtgctttta tctactccga tagcttccgc tattgctttt ttgctcatcg gtatttgcaa
     2041 catcatagaa attgcatacc tttgttcctc ggttatatgt ttgctcatct gcaacttttt
     2101 tttctttgga cggacaatta aagcaaagat agcaaacttt atccattcag agtgagagaa
     2161 agggggacat tgtctctctt tcctctctga aaaataaatg tttttattgc ttattatccg
     2221 cacccaaaaa gttgcattta taagttgaac tcaagaagta ttcacctgta agaagttact
     2281 aatgacaaaa aagaaattgc ccgttcgttt tacgggtcag cactttacta ttgataaagt
     2341 gctaataaaa gatgcaataa gacaagcaaa tataagtaat caggatacgg ttttagatat
     2401 tggggcaggc aaggggtttc ttactgttca tttattaaaa atcgccaaca atgttgttgc
     2461 tattgaaaac gacacagctt tggttgaaca tttacgaaaa ttattttctg atgcccgaaa
     2521 tgttcaagtt gtcggttgtg attttaggaa ttttgcagtt ccgaaatttc ctttcaaagt
     2581 ggtgtcaaat attccttatg gcattacttc cgatattttc aaaatcctga tgtttgagag
     2641 tcttggaaat tttctgggag gttccattgt ccttcaatta gaacctacac aaaagttatt
     2701 ttcgaggaag ctttacaatc catataccgt tttctatcat actttttttg atttgaaact
     2761 tgtctatgag gtaggtcctg aaagtttctt gccaccgcca actgtcaaat cagccctgtt
     2821 aaacattaaa agaaaacact tattttttga ttttaagttt aaagccaaat acttagcatt
     2881 tatttcctgt ctgttagaga aacctgattt atctgtaaaa acagctttaa agtcgatttt
     2941 caggaaaagt caggtcaggt caatttcgga aaaattcggt ttaaacctta atgctcaaat
     3001 tgtttgtttg tctccaagtc aatggttaaa ctgttttttg gaaatgctgg aagttgtccc
     3061 tgaaaaattt catccttcgt agtctagaca taaactgcca ggaattgggg atcggattcg
     3121 agctcgcacg ctgccgcaag cactcagggc gcaagggctg ctaaaggaag cggaacacgt
     3181 agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct
     3241 ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc
     3301 gatagctaga ctgggcggtc ttatggacag caagcgaacc ggaattgcca gctggggcgc
     3361 cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga
     3421 tctgatggcg caggggatca agatctgatc aagagacagg atactagtgg aggaagaaaa
     3481 aatggtgagc aagggcgagg cagtgatcaa ggagttcatg cggttcaagg tgcacatgga
     3541 gggctccatg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
     3601 gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct tctcctggga
     3661 catcctgtcc cctcagttca tgtacggctc cagggccttc atcaagcacc ccgccgacat
     3721 ccccgactac tataagcagt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     3781 cgaggacggc ggcgccgtga ccgtgaccca ggacacctcc ctggaggacg gcaccctgat
     3841 ctacaaggtg aagctccgcg gcaccaactt ccctcctgac ggccccgtaa tgcagaagaa
     3901 gacaatgggc tgggaagcgt ccaccgagcg gttgtacccc gaggacggcg tgctgaaggg
     3961 cgacattaag atggccctgc gcctgaagga cggcggccgc tacctggcgg acttcaagac
     4021 cacctacaag gccaagaagc ccgtgcagat gcccggcgcc tacaacgtcg accgcaagtt
     4081 ggacatcacc tcccacaacg aggactacac cgtggtggaa cagtacgaac gctccgaggg
     4141 ccgccactcc accggcggca tggacgagct gtacaagtaa gtaattagct gagcttggac
     4201 tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg
     4261 gttgccgccg ggcgtttttt attggtgaga atccaagcta gcttggcgag attttcagga
     4321 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
     4381 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
     4441 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
     4501 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg
     4561 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
     4621 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
     4681 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
     4741 aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt
     4801 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
     4861 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
     4921 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
     4981 cagggcgggg cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc tggggtaatg
     5041 actctctagc ttgaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     5101 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgctaggagc
     5161 ttgcggccgc cagcccgggc aattcgaggg gtatcgctct tgaagggaac tatgttgaat
     5221 aataaaaacg aattttcaca aaaaaatgtg tttttctttg ttagaccggg gacttatcag
     5281 ccaacctgtt atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5341 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5401 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5461 ctgcggccaa cttacttctg acaacgatcg aggcgattaa gttgggtaac gccagggttt
     5521 tcccagtcac gacgttgtaa aacgacggcc agtgaattaa ttcttgaaga cgaaagggcc
     5581 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagagcttat
     5641 cggccagcct cgcagagcag gattcccgtt gagcaccgcc aggtgcgaat aagggacagt
     5701 gaagaaggaa cacccgctcg cgggtgggcc tacttcacct atcctgcccg gctgacgccg
     5761 ttggatacac caaggaaagt ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa
     5821 aaaggatgga tataccgaaa aaatcgctat aatgaccccg aagcagggtt atgcagcggg
     5881 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     5941 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     6001 cacatatgga aaaaaaggaa tttcgtgttt tgataaaata ctgttttctg aagggaaaaa
     6061 atacagtgga agcaaaaact tggcttgata atgagtttcc ggactctgcc ccagggaaat
     6121 caacaataat tgattggtat gcaaaattca agcgtggtga aatgagcacg gaggacggtg
     6181 aacgcagtgg acgcccgaaa gaggtggtta ccgacgaaaa catcaaaaaa atccacaaaa
     6241 tgattttgaa tgaccgtaaa atgaagttga tcgagatagc agaggcctta aagatatcaa
     6301 aggaacgtgt tggtcatatc attcatcaat atttggatat gcggaagctc tgtgcaaaat
     6361 gggtgccgcg cgagctcaca tttgaccaaa aacaacaacg tgttgatgat tctgagcggt
     6421 gtttgcagct gttaactcgt aatacacccg agtttttccg tcgatatgtg acaatggatg
     6481 aaacatggct ccatcactac actcctgagt ccaatcgaca gtcggctgag tggacagcga
     6541 ccggtgaacc gtctccgaag cgtggaaaga ctcaaaagtc cgctggcaaa gtaatggcct
     6601 ctgttttttg ggatgcgcat ggaataattt ttatcgatta tcttgagaag ggaaaaacca
     6661 tcaacagtga ctattatatg gcgttattgg agcgtttgaa ggtcgaaatc gcggcaaaac
     6721 ggccccacat gaagaagaaa aaagtgttgt tccaccaaga caacgcaccg tgccacaagt
     6781 cattgagaac gatggcaaaa attcatgaat tgggcttcga attgcttccc cacccgccgt
     6841 attctccaga tctggccccc agcgactttt tcttgttctc agacctcaaa aggatgctcg
     6901 cagggaaaaa atttggctgc aatgaagagg tgatcgccga aactgaggcc tattttgagg
     6961 caaaaccgaa ggagtactac caaaatggta tcaaaaaatt ggaaggtcgt tataatcgtt
     7021 gtatcgctct tgaagggaac tatgttgaat aataaaaacg aattttcaca aaaaaatgtg
     7081 tttttctttg ttagaccgga tctgcagaat tccagcacac tggcggccgt ta
//
