LOCUS       sequence-378609-        6261 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 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-143
FEATURES             Location/Qualifiers
     source          1..6261
                     /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..2595,2596..3306)
                     /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(2635..2656)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2896..2915)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3243..3264
                     /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 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 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     2641 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     2701 gccacctacg gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc
     2761 tggcccaccc tcgtgaccac cctgggctac ggcgtgcagt gcttcgcccg ctaccccgac
     2821 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     2881 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2941 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     3001 ctggggcaca agctggagta caactacaac agccacaacg tctatatcac cgccgacaag
     3061 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg
     3121 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     3181 gacaaccact acctgagcta ccagtccaag ctgagcaaag accccaacga gaagcgcgat
     3241 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     3301 tacaagtaag 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
//
