LOCUS       sequence-383669-        6261 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 6261)
  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 6261)
  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-147
FEATURES             Location/Qualifiers
     source          1..6261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      588..977
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     promoter        join(1479..1484,1485..1501,1502..1507)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1696..2229
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     CDS             join(2591..2593,2594..2596,2597..3310)
                     /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(2636..2657)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     3244..3265
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      3327..3421
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3465..4124
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3531..3550)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      4189..4275
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4651..4673
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4665..4682
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4666..4682
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     oriT            4770..4878
                     /note="incP origin of transfer"
     protein_bind    5025..5046
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5061..5066,5067..5084,5085..5091)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5099..5115
                     /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     5104..5126
                     /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 aaaaagaata tataaggctt ttaaagcttt taaggtttaa cggttgtgga caacaagcca
      901 gggatgtaac gcactgagaa gcccttagag cctctcaaag caattttgag tgacacagga
      961 acacttaacg gctgacatgg gaattcttga agacgaaagg gcctcgtgat acgcctattt
     1021 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcggggt
     1081 taattaaggg ctgcaggaat tcgatatcaa gcttatcgat accgtcgaaa ttctgattaa
     1141 taatttgttt aaatttttcg tttggcgtga ggtatccaag tcttttacga ggtcgattat
     1201 tgagtttatt ttcaatccac ttaatctgtt tgttggttac ttcactaaag tccttaccct
     1261 ttgggatata ctgcctgata agcccgttgg tgttttcatt ggcaccacgt tcccatgagt
     1321 ggtatggttt gcaaaaatag aattttattt ccaatttttg cgcaatttcc tcgtgctcta
     1381 gaatataaac tgccaggaat tggggatcgg attcgagctt tagtacctag atttagatgt
     1441 ctaaaaagcg gggtaccgag gacgcgtcaa ttctcgaatt gacataagcc tgttcggttc
     1501 gtaaactgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
     1561 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgtac
     1621 agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt
     1681 tatggagcag caacgatgtt acgcagcagc aacgatgtta cgcagcaggg cagtcgccct
     1741 aaaacaaagt taggtggctc aagtatgggc atcattcgca catgtaggct cggccctgac
     1801 caagtcaaat ccatgcgggc tgctcttgat cttttcggtc gtgagttcgg agacgtagcc
     1861 acctactccc aacatcagcc ggactccgat tacctcggga acttgctccg tagtaagaca
     1921 ttcatcgcgc ttgctgcctt cgaccaagaa gcggttgttg gcgctctcgc ggcttacgtt
     1981 ctgcccaagt ttgagcagcc gcgtagtgag atctatatct atgatctcgc agtctccggc
     2041 gagcaccgga ggcagggcat tgccaccgcg ctcatcaatc tcctcaagca tgaggccaac
     2101 gcgcttggtg cttatgtgat ctacgtgcaa gcagattacg gtgacgatcc cgcagtggct
     2161 ctctatacaa agttgggcat acgggaagaa gtgatgcact ttgatatcga cccaagtacc
     2221 gccacctaaa gctcgcacgc tgccgcaagc actcagggcg caagggctgc taaaggaagc
     2281 ggaacacgta gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact
     2341 gggctatctg gacaagggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc
     2401 ttacatggcg atagctagac tgggcggttt tatggacagc aagcgaaccg gaattgccag
     2461 ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc
     2521 cgccaaggat ctgatggcgc aggggatcaa gatctgatca agagacagga tactagtgga
     2581 ggaagaaaaa atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     2641 cgagctggac ggcgacgtaa acggccacaa gttcagcgtc cgcggcgagg gcgagggcga
     2701 tgccaccaac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     2761 ctggcccacc ctcgtgacca ccttcggcta cggcgtggcc tgcttcagcc gctaccccga
     2821 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     2881 caccatctct ttcaaggacg acggtaccta caagacccgc gccgaggtga agttcgaggg
     2941 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     3001 cctggggcac aagctggagt acaacttcaa cagccactac gtctatatca cggccgacaa
     3061 gcagaagaac tgcatcaagg ctaacttcaa gatccgccac aacgttgagg acggcagcgt
     3121 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     3181 cgacaaccac tacctgagcc atcagtccaa gctgagcaaa gaccccaacg agaagcgcga
     3241 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatt acacatggca tggacgagct
     3301 gtacaagtaa taattagctg agcttggact cctgttgata gatccagtaa tgacctcaga
     3361 actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa
     3421 tccaagctag cttggcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca
     3481 ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc
     3541 agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa
     3601 agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc
     3661 tgatgaatgc tcatccggaa tttcgtatgg caatgaaaga cggtgagctg gtgatatggg
     3721 atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct
     3781 ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt
     3841 gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct
     3901 cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact
     3961 tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc
     4021 cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta
     4081 atgaattaca acagtactgc gatgagtggc agggcggggc gtaatttttt taaggcagtt
     4141 attggtgccc ttaaacgcct ggggtaatga ctctctagct tgaggcatca aataaaacga
     4201 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     4261 ctgagtagga caaatccgcc gctaggagct tgcggccgcc agcccgggca attcgagggg
     4321 tatcgctctt gaagggaact atgttgaata ataaaaacga attttcacaa aaaaatgtgt
     4381 ttttctttgt tagaccgggg acttatcagc caacctgtta tgtggcgcgg tattatcccg
     4441 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4501 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4561 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcga
     4621 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     4681 gtgaattaat tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4741 catgataata atggtttctt agagcttatc ggccagcctc gcagagcagg attcccgttg
     4801 agcaccgcca ggtgcgaata agggacagtg aagaaggaac acccgctcgc gggtgggcct
     4861 acttcaccta tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc tacacgaacc
     4921 ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata
     4981 atgaccccga agcagggtta tgcagcgggt gagcgcaacg caattaatgt gagttagctc
     5041 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     5101 gtgagcggat aacaatttca cacaggaaac acatatggaa aaaaaggaat ttcgtgtttt
     5161 gataaaatac tgttttctga agggaaaaaa tacagtggaa gcaaaaactt ggcttgataa
     5221 tgagtttccg gactctgccc cagggaaatc aacaataatt gattggtatg caaaattcaa
     5281 gcgtggtgaa atgagcacgg aggacggtga acgcagtgga cgcccgaaag aggtggttac
     5341 cgacgaaaac atcaaaaaaa tccacaaaat gattttgaat gaccgtaaaa tgaagttgat
     5401 cgagatagca gaggccttaa agatatcaaa ggaacgtgtt ggtcatatca ttcatcaata
     5461 tttggatatg cggaagctct gtgcaaaatg ggtgccgcgc gagctcacat ttgaccaaaa
     5521 acaacaacgt gttgatgatt ctgagcggtg tttgcagctg ttaactcgta atacacccga
     5581 gtttttccgt cgatatgtga caatggatga aacatggctc catcactaca ctcctgagtc
     5641 caatcgacag tcggctgagt ggacagcgac cggtgaaccg tctccgaagc gtggaaagac
     5701 tcaaaagtcc gctggcaaag taatggcctc tgttttttgg gatgcgcatg gaataatttt
     5761 tatcgattat cttgagaagg gaaaaaccat caacagtgac tattatatgg cgttattgga
     5821 gcgtttgaag gtcgaaatcg cggcaaaacg gccccacatg aagaagaaaa aagtgttgtt
     5881 ccaccaagac aacgcaccgt gccacaagtc attgagaacg atggcaaaaa ttcatgaatt
     5941 gggcttcgaa ttgcttcccc acccgccgta ttctccagat ctggccccca gcgacttttt
     6001 cttgttctca gacctcaaaa ggatgctcgc agggaaaaaa tttggctgca atgaagaggt
     6061 gatcgccgaa actgaggcct attttgaggc aaaaccgaag gagtactacc aaaatggtat
     6121 caaaaaattg gaaggtcgtt ataatcgttg tatcgctctt gaagggaact atgttgaata
     6181 ataaaaacga attttcacaa aaaaatgtgt ttttctttgt tagaccggat ctgcagaatt
     6241 ccagcacact ggcggccgtt a
//
