LOCUS       sequence-377499-        5447 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Himar transposon delivery plasmid that delivers a yellow fluorescent
            sYFP2 protein gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5447)
  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 5447)
  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-133
FEATURES             Location/Qualifiers
     source          1..5447
                     /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(1776..1778,1779..1781,1782..2492)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1821..1842)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2082..2101)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2429..2450
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      2513..2607
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2651..3310
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2717..2736)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3375..3461
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3837..3859
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3851..3868
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3852..3868
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     oriT            3956..4064
                     /note="incP origin of transfer"
     protein_bind    4211..4232
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4247..4252,4253..4270,4271..4277)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4285..4301
                     /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     4290..4312
                     /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 gaggagctgt
     1801 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     1861 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct
     1921 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcg
     1981 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2041 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     2101 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     2161 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     2221 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
     2281 gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca
     2341 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga
     2401 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     2461 ggatcactct cggcatggac gagctgtaca agtaagtaat tagctgagct tggactcctg
     2521 ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc
     2581 cgccgggcgt tttttattgg tgagaatcca agctagcttg gcgagatttt caggagctaa
     2641 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca
     2701 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt
     2761 tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
     2821 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaatttc gtatggcaat
     2881 gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga
     2941 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct
     3001 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg
     3061 gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga
     3121 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta
     3181 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     3241 tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg
     3301 cggggcgtaa tttttttaag gcagttattg gtgcccttaa acgcctgggg taatgactct
     3361 ctagcttgag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     3421 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccgcta ggagcttgcg
     3481 gccgccagcc cgggcaattc gaggggtatc gctcttgaag ggaactatgt tgaataataa
     3541 aaacgaattt tcacaaaaaa atgtgttttt ctttgttaga ccggggactt atcagccaac
     3601 ctgttatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     3661 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3721 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3781 gccaacttac ttctgacaac gatcgaggcg attaagttgg gtaacgccag ggttttccca
     3841 gtcacgacgt tgtaaaacga cggccagtga attaattctt gaagacgaaa gggcctcgtg
     3901 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagag cttatcggcc
     3961 agcctcgcag agcaggattc ccgttgagca ccgccaggtg cgaataaggg acagtgaaga
     4021 aggaacaccc gctcgcgggt gggcctactt cacctatcct gcccggctga cgccgttgga
     4081 tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt gcgaaaaagg
     4141 atggatatac cgaaaaaatc gctataatga ccccgaagca gggttatgca gcgggtgagc
     4201 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     4261 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacacat
     4321 atggaaaaaa aggaatttcg tgttttgata aaatactgtt ttctgaaggg aaaaaataca
     4381 gtggaagcaa aaacttggct tgataatgag tttccggact ctgccccagg gaaatcaaca
     4441 ataattgatt ggtatgcaaa attcaagcgt ggtgaaatga gcacggagga cggtgaacgc
     4501 agtggacgcc cgaaagaggt ggttaccgac gaaaacatca aaaaaatcca caaaatgatt
     4561 ttgaatgacc gtaaaatgaa gttgatcgag atagcagagg ccttaaagat atcaaaggaa
     4621 cgtgttggtc atatcattca tcaatatttg gatatgcgga agctctgtgc aaaatgggtg
     4681 ccgcgcgagc tcacatttga ccaaaaacaa caacgtgttg atgattctga gcggtgtttg
     4741 cagctgttaa ctcgtaatac acccgagttt ttccgtcgat atgtgacaat ggatgaaaca
     4801 tggctccatc actacactcc tgagtccaat cgacagtcgg ctgagtggac agcgaccggt
     4861 gaaccgtctc cgaagcgtgg aaagactcaa aagtccgctg gcaaagtaat ggcctctgtt
     4921 ttttgggatg cgcatggaat aatttttatc gattatcttg agaagggaaa aaccatcaac
     4981 agtgactatt atatggcgtt attggagcgt ttgaaggtcg aaatcgcggc aaaacggccc
     5041 cacatgaaga agaaaaaagt gttgttccac caagacaacg caccgtgcca caagtcattg
     5101 agaacgatgg caaaaattca tgaattgggc ttcgaattgc ttccccaccc gccgtattct
     5161 ccagatctgg cccccagcga ctttttcttg ttctcagacc tcaaaaggat gctcgcaggg
     5221 aaaaaatttg gctgcaatga agaggtgatc gccgaaactg aggcctattt tgaggcaaaa
     5281 ccgaaggagt actaccaaaa tggtatcaaa aaattggaag gtcgttataa tcgttgtatc
     5341 gctcttgaag ggaactatgt tgaataataa aaacgaattt tcacaaaaaa atgtgttttt
     5401 ctttgttaga ccggatctgc agaattccag cacactggcg gccgtta
//
