LOCUS       sequence-378616-        6300 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Himar transposon delivery plasmid that delivers a green fluorescent 
            mClover3 protein gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6300)
  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 6300)
  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-157
FEATURES             Location/Qualifiers
     source          1..6300
                     /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             join(2630..2632,2633..2635,2636..3349)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(2675..2696)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     3283..3304
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      3366..3460
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3504..4163
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3570..3589)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      4228..4314
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4690..4712
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4704..4721
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4705..4721
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     oriT            4809..4917
                     /note="incP origin of transfer"
     protein_bind    5064..5085
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5100..5105,5106..5123,5124..5130)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5138..5154
                     /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     5143..5165
                     /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 gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc tctggtaagg
     2521 ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca
     2581 ggggatcaag atctgatcaa gagacaggat actagtggag gaagaaaaaa tggtgagcaa
     2641 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     2701 cggccacaag ttcagcgtcc gcggcgaggg cgagggcgat gccaccaacg gcaagctgac
     2761 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     2821 cttcggctac ggcgtggcct gcttcagccg ctaccccgac cacatgaagc agcacgactt
     2881 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatctctt tcaaggacga
     2941 cggtacctac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     3001 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     3061 caacttcaac agccactacg tctatatcac ggccgacaag cagaagaact gcatcaaggc
     3121 taacttcaag atccgccaca acgttgagga cggcagcgtg cagctcgccg accactacca
     3181 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcca
     3241 tcagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     3301 cgtgaccgcc gccgggatta cacatggcat ggacgagctg tacaagtaat aattagctga
     3361 gcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg atttgttcag
     3421 aacgctcggt tgccgccggg cgttttttat tggtgagaat ccaagctagc ttggcgagat
     3481 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
     3541 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
     3601 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
     3661 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
     3721 ttcgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
     3781 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
     3841 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
     3901 atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt
     3961 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
     4021 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
     4081 atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
     4141 atgagtggca gggcggggcg taattttttt aaggcagtta ttggtgccct taaacgcctg
     4201 gggtaatgac tctctagctt gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     4261 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     4321 ctaggagctt gcggccgcca gcccgggcaa ttcgaggggt atcgctcttg aagggaacta
     4381 tgttgaataa taaaaacgaa ttttcacaaa aaaatgtgtt tttctttgtt agaccgggga
     4441 cttatcagcc aacctgttat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4501 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4561 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4621 tgataacact gcggccaact tacttctgac aacgatcgag gcgattaagt tgggtaacgc
     4681 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattaatt cttgaagacg
     4741 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4801 gagcttatcg gccagcctcg cagagcagga ttcccgttga gcaccgccag gtgcgaataa
     4861 gggacagtga agaaggaaca cccgctcgcg ggtgggccta cttcacctat cctgcccggc
     4921 tgacgccgtt ggatacacca aggaaagtct acacgaaccc tttggcaaaa tcctgtatat
     4981 cgtgcgaaaa aggatggata taccgaaaaa atcgctataa tgaccccgaa gcagggttat
     5041 gcagcgggtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     5101 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     5161 acaggaaaca catatggaaa aaaaggaatt tcgtgttttg ataaaatact gttttctgaa
     5221 gggaaaaaat acagtggaag caaaaacttg gcttgataat gagtttccgg actctgcccc
     5281 agggaaatca acaataattg attggtatgc aaaattcaag cgtggtgaaa tgagcacgga
     5341 ggacggtgaa cgcagtggac gcccgaaaga ggtggttacc gacgaaaaca tcaaaaaaat
     5401 ccacaaaatg attttgaatg accgtaaaat gaagttgatc gagatagcag aggccttaaa
     5461 gatatcaaag gaacgtgttg gtcatatcat tcatcaatat ttggatatgc ggaagctctg
     5521 tgcaaaatgg gtgccgcgcg agctcacatt tgaccaaaaa caacaacgtg ttgatgattc
     5581 tgagcggtgt ttgcagctgt taactcgtaa tacacccgag tttttccgtc gatatgtgac
     5641 aatggatgaa acatggctcc atcactacac tcctgagtcc aatcgacagt cggctgagtg
     5701 gacagcgacc ggtgaaccgt ctccgaagcg tggaaagact caaaagtccg ctggcaaagt
     5761 aatggcctct gttttttggg atgcgcatgg aataattttt atcgattatc ttgagaaggg
     5821 aaaaaccatc aacagtgact attatatggc gttattggag cgtttgaagg tcgaaatcgc
     5881 ggcaaaacgg ccccacatga agaagaaaaa agtgttgttc caccaagaca acgcaccgtg
     5941 ccacaagtca ttgagaacga tggcaaaaat tcatgaattg ggcttcgaat tgcttcccca
     6001 cccgccgtat tctccagatc tggcccccag cgactttttc ttgttctcag acctcaaaag
     6061 gatgctcgca gggaaaaaat ttggctgcaa tgaagaggtg atcgccgaaa ctgaggccta
     6121 ttttgaggca aaaccgaagg agtactacca aaatggtatc aaaaaattgg aaggtcgtta
     6181 taatcgttgt atcgctcttg aagggaacta tgttgaataa taaaaacgaa ttttcacaaa
     6241 aaaatgtgtt tttctttgtt agaccggatc tgcagaattc cagcacactg gcggccgtta
//
