LOCUS       sequence-383668-        6224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Himar transposon delivery plasmid that delivers a green fluorescent 
            sGFP2 protein gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6224)
  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 6224)
  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-142
FEATURES             Location/Qualifiers
     source          1..6224
                     /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"
     promoter        join(1478..1483,1484..1500,1501..1506)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1695..2228
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     CDS             join(2590..2592,2593..3105)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIE"
     primer_bind     complement(2632..2653)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2893..2912)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3240..3261
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      3290..3384
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3428..4087
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3494..3513)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      4152..4238
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4614..4636
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4628..4645
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4629..4645
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     oriT            4733..4841
                     /note="incP origin of transfer"
     protein_bind    4988..5009
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5024..5029,5030..5047,5048..5054)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5062..5078
                     /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     5067..5089
                     /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 ttcgagcttt agtacctaga tttagatgtc
     1441 taaaaagcgg ggtaccgagg acgcgtcaat tctcgaattg acataagcct gttcggttcg
     1501 taaactgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg
     1561 cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttgtaca
     1621 gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt
     1681 atggagcagc aacgatgtta cgcagcagca acgatgttac gcagcagggc agtcgcccta
     1741 aaacaaagtt aggtggctca agtatgggca tcattcgcac atgtaggctc ggccctgacc
     1801 aagtcaaatc catgcgggct gctcttgatc ttttcggtcg tgagttcgga gacgtagcca
     1861 cctactccca acatcagccg gactccgatt acctcgggaa cttgctccgt agtaagacat
     1921 tcatcgcgct tgctgccttc gaccaagaag cggttgttgg cgctctcgcg gcttacgttc
     1981 tgcccaagtt tgagcagccg cgtagtgaga tctatatcta tgatctcgca gtctccggcg
     2041 agcaccggag gcagggcatt gccaccgcgc tcatcaatct cctcaagcat gaggccaacg
     2101 cgcttggtgc ttatgtgatc tacgtgcaag cagattacgg tgacgatccc gcagtggctc
     2161 tctatacaaa gttgggcata cgggaagaag tgatgcactt tgatatcgac ccaagtaccg
     2221 ccacctaaag ctcgcacgct gccgcaagca ctcagggcgc aagggctgct aaaggaagcg
     2281 gaacacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg
     2341 ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct
     2401 tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc
     2461 tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc
     2521 gccaaggatc tgatggcgca ggggatcaag atctgatcaa gagacaggat actagtggag
     2581 gaagaaaaaa tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2641 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     2701 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     2761 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcgcccgcta ccccgaccac
     2821 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     2881 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     2941 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     3001 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag
     3061 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     3121 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3181 aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
     3241 atggtcctgc tggagttcgt gaccgccgcc gggtaattag ctgagcttgg actcctgttg
     3301 atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc
     3361 cgggcgtttt ttattggtga gaatccaagc tagcttggcg agattttcag gagctaagga
     3421 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
     3481 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
     3541 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
     3601 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa
     3661 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
     3721 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
     3781 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
     3841 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
     3901 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
     3961 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg
     4021 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
     4081 ggcgtaattt ttttaaggca gttattggtg cccttaaacg cctggggtaa tgactctcta
     4141 gcttgaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     4201 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgctagga gcttgcggcc
     4261 gccagcccgg gcaattcgag gggtatcgct cttgaaggga actatgttga ataataaaaa
     4321 cgaattttca caaaaaaatg tgtttttctt tgttagaccg gggacttatc agccaacctg
     4381 ttatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4441 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4501 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4561 aacttacttc tgacaacgat cgaggcgatt aagttgggta acgccagggt tttcccagtc
     4621 acgacgttgt aaaacgacgg ccagtgaatt aattcttgaa gacgaaaggg cctcgtgata
     4681 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagagctt atcggccagc
     4741 ctcgcagagc aggattcccg ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg
     4801 aacacccgct cgcgggtggg cctacttcac ctatcctgcc cggctgacgc cgttggatac
     4861 accaaggaaa gtctacacga accctttggc aaaatcctgt atatcgtgcg aaaaaggatg
     4921 gatataccga aaaaatcgct ataatgaccc cgaagcaggg ttatgcagcg ggtgagcgca
     4981 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     5041 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacacatatg
     5101 gaaaaaaagg aatttcgtgt tttgataaaa tactgttttc tgaagggaaa aaatacagtg
     5161 gaagcaaaaa cttggcttga taatgagttt ccggactctg ccccagggaa atcaacaata
     5221 attgattggt atgcaaaatt caagcgtggt gaaatgagca cggaggacgg tgaacgcagt
     5281 ggacgcccga aagaggtggt taccgacgaa aacatcaaaa aaatccacaa aatgattttg
     5341 aatgaccgta aaatgaagtt gatcgagata gcagaggcct taaagatatc aaaggaacgt
     5401 gttggtcata tcattcatca atatttggat atgcggaagc tctgtgcaaa atgggtgccg
     5461 cgcgagctca catttgacca aaaacaacaa cgtgttgatg attctgagcg gtgtttgcag
     5521 ctgttaactc gtaatacacc cgagtttttc cgtcgatatg tgacaatgga tgaaacatgg
     5581 ctccatcact acactcctga gtccaatcga cagtcggctg agtggacagc gaccggtgaa
     5641 ccgtctccga agcgtggaaa gactcaaaag tccgctggca aagtaatggc ctctgttttt
     5701 tgggatgcgc atggaataat ttttatcgat tatcttgaga agggaaaaac catcaacagt
     5761 gactattata tggcgttatt ggagcgtttg aaggtcgaaa tcgcggcaaa acggccccac
     5821 atgaagaaga aaaaagtgtt gttccaccaa gacaacgcac cgtgccacaa gtcattgaga
     5881 acgatggcaa aaattcatga attgggcttc gaattgcttc cccacccgcc gtattctcca
     5941 gatctggccc ccagcgactt tttcttgttc tcagacctca aaaggatgct cgcagggaaa
     6001 aaatttggct gcaatgaaga ggtgatcgcc gaaactgagg cctattttga ggcaaaaccg
     6061 aaggagtact accaaaatgg tatcaaaaaa ttggaaggtc gttataatcg ttgtatcgct
     6121 cttgaaggga actatgttga ataataaaaa cgaattttca caaaaaaatg tgtttttctt
     6181 tgttagaccg gatctgcaga attccagcac actggcggcc gtta
//
