LOCUS       sequence-377500-        5441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Himar transposon delivery plasmid that delivers a cyan fluorescent 
            mTurquoise2 protein gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5441)
  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 5441)
  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-131
FEATURES             Location/Qualifiers
     source          1..5441
                     /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             join(1777..1779,1780..2493)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1819..1840)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2080..2099)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2427..2448
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      2507..2601
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2645..3304
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2711..2730)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3369..3455
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3831..3853
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3845..3862
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3846..3862
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     oriT            3950..4058
                     /note="incP origin of transfer"
     protein_bind    4205..4226
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4241..4246,4247..4264,4265..4271)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4279..4295
                     /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     4284..4306
                     /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 ggttttatgg
     1621 acagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc
     1681 aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagatct
     1741 gatcaagaga caggatacta gtggaggaag aaaaaaatga gcaagggcga ggagctgttc
     1801 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1861 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1921 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgtc ctggggcgtg
     1981 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2041 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2101 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2161 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta ctttagcgac
     2221 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     2281 cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc
     2341 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc
     2401 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2461 atcactctcg gcatggacga gctgtacaag taattagctg agcttggact cctgttgata
     2521 gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg
     2581 gcgtttttta ttggtgagaa tccaagctag cttggcgaga ttttcaggag ctaaggaagc
     2641 taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa
     2701 agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct
     2761 ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt
     2821 tattcacatt cttgcccgcc tgatgaatgc tcatccggaa tttcgtatgg caatgaaaga
     2881 cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac
     2941 tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat
     3001 atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat
     3061 tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa
     3121 cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca
     3181 aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt
     3241 ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc
     3301 gtaatttttt taaggcagtt attggtgccc ttaaacgcct ggggtaatga ctctctagct
     3361 tgaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     3421 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gctaggagct tgcggccgcc
     3481 agcccgggca attcgagggg tatcgctctt gaagggaact atgttgaata ataaaaacga
     3541 attttcacaa aaaaatgtgt ttttctttgt tagaccgggg acttatcagc caacctgtta
     3601 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     3661 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3721 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3781 ttacttctga caacgatcga ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     3841 acgttgtaaa acgacggcca gtgaattaat tcttgaagac gaaagggcct cgtgatacgc
     3901 ctatttttat aggttaatgt catgataata atggtttctt agagcttatc ggccagcctc
     3961 gcagagcagg attcccgttg agcaccgcca ggtgcgaata agggacagtg aagaaggaac
     4021 acccgctcgc gggtgggcct acttcaccta tcctgcccgg ctgacgccgt tggatacacc
     4081 aaggaaagtc tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat
     4141 ataccgaaaa aatcgctata atgaccccga agcagggtta tgcagcgggt gagcgcaacg
     4201 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     4261 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac acatatggaa
     4321 aaaaaggaat ttcgtgtttt gataaaatac tgttttctga agggaaaaaa tacagtggaa
     4381 gcaaaaactt ggcttgataa tgagtttccg gactctgccc cagggaaatc aacaataatt
     4441 gattggtatg caaaattcaa gcgtggtgaa atgagcacgg aggacggtga acgcagtgga
     4501 cgcccgaaag aggtggttac cgacgaaaac atcaaaaaaa tccacaaaat gattttgaat
     4561 gaccgtaaaa tgaagttgat cgagatagca gaggccttaa agatatcaaa ggaacgtgtt
     4621 ggtcatatca ttcatcaata tttggatatg cggaagctct gtgcaaaatg ggtgccgcgc
     4681 gagctcacat ttgaccaaaa acaacaacgt gttgatgatt ctgagcggtg tttgcagctg
     4741 ttaactcgta atacacccga gtttttccgt cgatatgtga caatggatga aacatggctc
     4801 catcactaca ctcctgagtc caatcgacag tcggctgagt ggacagcgac cggtgaaccg
     4861 tctccgaagc gtggaaagac tcaaaagtcc gctggcaaag taatggcctc tgttttttgg
     4921 gatgcgcatg gaataatttt tatcgattat cttgagaagg gaaaaaccat caacagtgac
     4981 tattatatgg cgttattgga gcgtttgaag gtcgaaatcg cggcaaaacg gccccacatg
     5041 aagaagaaaa aagtgttgtt ccaccaagac aacgcaccgt gccacaagtc attgagaacg
     5101 atggcaaaaa ttcatgaatt gggcttcgaa ttgcttcccc acccgccgta ttctccagat
     5161 ctggccccca gcgacttttt cttgttctca gacctcaaaa ggatgctcgc agggaaaaaa
     5221 tttggctgca atgaagaggt gatcgccgaa actgaggcct attttgaggc aaaaccgaag
     5281 gagtactacc aaaatggtat caaaaaattg gaaggtcgtt ataatcgttg tatcgctctt
     5341 gaagggaact atgttgaata ataaaaacga attttcacaa aaaaatgtgt ttttctttgt
     5401 tagaccggat ctgcagaatt ccagcacact ggcggccgtt a
//
