LOCUS       sequence-377504-        5437 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Himar transposon delivery plasmid that delivers a orange fluorescent
            mOrange2 protein gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5437)
  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 5437)
  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-134
FEATURES             Location/Qualifiers
     source          1..5437
                     /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..2486
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     terminator      2503..2597
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2641..3300
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2707..2726)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3365..3451
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3827..3849
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3841..3858
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3842..3858
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     oriT            3946..4054
                     /note="incP origin of transfer"
     protein_bind    4201..4222
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4237..4242,4243..4260,4261..4267)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4275..4291
                     /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     4280..4302
                     /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 aaaaaatggt gagcaagggc gaggagaata
     1801 acatggccat catcaaggag ttcatgcgct tcaaggtgcg catggagggc tccgtgaacg
     1861 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc tttcagaccg
     1921 ctaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     1981 atttcaccta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttca
     2041 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaactacgag gacggcggcg
     2101 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     2161 tgcgcggcac caacttcccc tccgacggcc ccgtgatgca gaagaagacc atgggctggg
     2221 aggcctcctc cgagcggatg taccccgagg acggtgccct gaagggcaag atcaagatga
     2281 ggctgaagct gaaggacggc ggccactaca cctccgaggt caagaccacc tacaaggcca
     2341 agaagcccgt gcagctgccc ggcgcctaca tcgtcgacat caagttggac atcacctccc
     2401 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     2461 gcggcatgga cgagctgtac aagtaataat tagctgagct tggactcctg ttgatagatc
     2521 cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt
     2581 tttttattgg tgagaatcca agctagcttg gcgagatttt caggagctaa ggaagctaaa
     2641 atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa
     2701 cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat
     2761 attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt
     2821 cacattcttg cccgcctgat gaatgctcat ccggaatttc gtatggcaat gaaagacggt
     2881 gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa
     2941 acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat
     3001 tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag
     3061 aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg
     3121 gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc
     3181 gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat
     3241 gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa
     3301 tttttttaag gcagttattg gtgcccttaa acgcctgggg taatgactct ctagcttgag
     3361 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     3421 gtcggtgaac gctctcctga gtaggacaaa tccgccgcta ggagcttgcg gccgccagcc
     3481 cgggcaattc gaggggtatc gctcttgaag ggaactatgt tgaataataa aaacgaattt
     3541 tcacaaaaaa atgtgttttt ctttgttaga ccggggactt atcagccaac ctgttatgtg
     3601 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3661 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3721 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3781 ttctgacaac gatcgaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     3841 tgtaaaacga cggccagtga attaattctt gaagacgaaa gggcctcgtg atacgcctat
     3901 ttttataggt taatgtcatg ataataatgg tttcttagag cttatcggcc agcctcgcag
     3961 agcaggattc ccgttgagca ccgccaggtg cgaataaggg acagtgaaga aggaacaccc
     4021 gctcgcgggt gggcctactt cacctatcct gcccggctga cgccgttgga tacaccaagg
     4081 aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac
     4141 cgaaaaaatc gctataatga ccccgaagca gggttatgca gcgggtgagc gcaacgcaat
     4201 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     4261 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacacat atggaaaaaa
     4321 aggaatttcg tgttttgata aaatactgtt ttctgaaggg aaaaaataca gtggaagcaa
     4381 aaacttggct tgataatgag tttccggact ctgccccagg gaaatcaaca ataattgatt
     4441 ggtatgcaaa attcaagcgt ggtgaaatga gcacggagga cggtgaacgc agtggacgcc
     4501 cgaaagaggt ggttaccgac gaaaacatca aaaaaatcca caaaatgatt ttgaatgacc
     4561 gtaaaatgaa gttgatcgag atagcagagg ccttaaagat atcaaaggaa cgtgttggtc
     4621 atatcattca tcaatatttg gatatgcgga agctctgtgc aaaatgggtg ccgcgcgagc
     4681 tcacatttga ccaaaaacaa caacgtgttg atgattctga gcggtgtttg cagctgttaa
     4741 ctcgtaatac acccgagttt ttccgtcgat atgtgacaat ggatgaaaca tggctccatc
     4801 actacactcc tgagtccaat cgacagtcgg ctgagtggac agcgaccggt gaaccgtctc
     4861 cgaagcgtgg aaagactcaa aagtccgctg gcaaagtaat ggcctctgtt ttttgggatg
     4921 cgcatggaat aatttttatc gattatcttg agaagggaaa aaccatcaac agtgactatt
     4981 atatggcgtt attggagcgt ttgaaggtcg aaatcgcggc aaaacggccc cacatgaaga
     5041 agaaaaaagt gttgttccac caagacaacg caccgtgcca caagtcattg agaacgatgg
     5101 caaaaattca tgaattgggc ttcgaattgc ttccccaccc gccgtattct ccagatctgg
     5161 cccccagcga ctttttcttg ttctcagacc tcaaaaggat gctcgcaggg aaaaaatttg
     5221 gctgcaatga agaggtgatc gccgaaactg aggcctattt tgaggcaaaa ccgaaggagt
     5281 actaccaaaa tggtatcaaa aaattggaag gtcgttataa tcgttgtatc gctcttgaag
     5341 ggaactatgt tgaataataa aaacgaattt tcacaaaaaa atgtgttttt ctttgttaga
     5401 ccggatctgc agaattccag cacactggcg gccgtta
//
