LOCUS       sequence-378612-        6240 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Himar transposon delivery plasmid that delivers a red fluorescent 
            mScarlet-I protein gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6240)
  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 6240)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-145
FEATURES             Location/Qualifiers
     source          1..6240
                     /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             2590..3285
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(2723..2741)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2991..3010
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      3306..3400
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3444..4103
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3510..3529)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      4168..4254
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4630..4652
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4644..4661
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4645..4661
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     oriT            4749..4857
                     /note="incP origin of transfer"
     protein_bind    5004..5025
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5040..5045,5046..5063,5064..5070)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5078..5094
                     /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     5083..5105
                     /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 gggcggtctt atggacagca agcgaaccgg aattgccagc
     2461 tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc
     2521 gccaaggatc tgatggcgca ggggatcaag atctgatcaa gagacaggat actagtggag
     2581 gaagaaaaaa tggtgagcaa gggcgaggca gtgatcaagg agttcatgcg gttcaaggtg
     2641 cacatggagg gctccatgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     2701 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     2761 tcctgggaca tcctgtcccc tcagttcatg tacggctcca gggccttcat caagcacccc
     2821 gccgacatcc ccgactacta taagcagtcc ttccccgagg gcttcaagtg ggagcgcgtg
     2881 atgaacttcg aggacggcgg cgccgtgacc gtgacccagg acacctccct ggaggacggc
     2941 accctgatct acaaggtgaa gctccgcggc accaacttcc ctcctgacgg ccccgtaatg
     3001 cagaagaaga caatgggctg ggaagcgtcc accgagcggt tgtaccccga ggacggcgtg
     3061 ctgaagggcg acattaagat ggccctgcgc ctgaaggacg gcggccgcta cctggcggac
     3121 ttcaagacca cctacaaggc caagaagccc gtgcagatgc ccggcgccta caacgtcgac
     3181 cgcaagttgg acatcacctc ccacaacgag gactacaccg tggtggaaca gtacgaacgc
     3241 tccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaagt aattagctga
     3301 gcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg atttgttcag
     3361 aacgctcggt tgccgccggg cgttttttat tggtgagaat ccaagctagc ttggcgagat
     3421 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
     3481 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
     3541 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
     3601 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
     3661 ttcgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
     3721 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
     3781 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
     3841 atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt
     3901 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
     3961 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
     4021 atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
     4081 atgagtggca gggcggggcg taattttttt aaggcagtta ttggtgccct taaacgcctg
     4141 gggtaatgac tctctagctt gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     4201 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     4261 ctaggagctt gcggccgcca gcccgggcaa ttcgaggggt atcgctcttg aagggaacta
     4321 tgttgaataa taaaaacgaa ttttcacaaa aaaatgtgtt tttctttgtt agaccgggga
     4381 cttatcagcc aacctgttat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4441 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4501 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4561 tgataacact gcggccaact tacttctgac aacgatcgag gcgattaagt tgggtaacgc
     4621 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattaatt cttgaagacg
     4681 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4741 gagcttatcg gccagcctcg cagagcagga ttcccgttga gcaccgccag gtgcgaataa
     4801 gggacagtga agaaggaaca cccgctcgcg ggtgggccta cttcacctat cctgcccggc
     4861 tgacgccgtt ggatacacca aggaaagtct acacgaaccc tttggcaaaa tcctgtatat
     4921 cgtgcgaaaa aggatggata taccgaaaaa atcgctataa tgaccccgaa gcagggttat
     4981 gcagcgggtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     5041 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     5101 acaggaaaca catatggaaa aaaaggaatt tcgtgttttg ataaaatact gttttctgaa
     5161 gggaaaaaat acagtggaag caaaaacttg gcttgataat gagtttccgg actctgcccc
     5221 agggaaatca acaataattg attggtatgc aaaattcaag cgtggtgaaa tgagcacgga
     5281 ggacggtgaa cgcagtggac gcccgaaaga ggtggttacc gacgaaaaca tcaaaaaaat
     5341 ccacaaaatg attttgaatg accgtaaaat gaagttgatc gagatagcag aggccttaaa
     5401 gatatcaaag gaacgtgttg gtcatatcat tcatcaatat ttggatatgc ggaagctctg
     5461 tgcaaaatgg gtgccgcgcg agctcacatt tgaccaaaaa caacaacgtg ttgatgattc
     5521 tgagcggtgt ttgcagctgt taactcgtaa tacacccgag tttttccgtc gatatgtgac
     5581 aatggatgaa acatggctcc atcactacac tcctgagtcc aatcgacagt cggctgagtg
     5641 gacagcgacc ggtgaaccgt ctccgaagcg tggaaagact caaaagtccg ctggcaaagt
     5701 aatggcctct gttttttggg atgcgcatgg aataattttt atcgattatc ttgagaaggg
     5761 aaaaaccatc aacagtgact attatatggc gttattggag cgtttgaagg tcgaaatcgc
     5821 ggcaaaacgg ccccacatga agaagaaaaa agtgttgttc caccaagaca acgcaccgtg
     5881 ccacaagtca ttgagaacga tggcaaaaat tcatgaattg ggcttcgaat tgcttcccca
     5941 cccgccgtat tctccagatc tggcccccag cgactttttc ttgttctcag acctcaaaag
     6001 gatgctcgca gggaaaaaat ttggctgcaa tgaagaggtg atcgccgaaa ctgaggccta
     6061 ttttgaggca aaaccgaagg agtactacca aaatggtatc aaaaaattgg aaggtcgtta
     6121 taatcgttgt atcgctcttg aagggaacta tgttgaataa taaaaacgaa ttttcacaaa
     6181 aaaatgtgtt tttctttgtt agaccggatc tgcagaattc cagcacactg gcggccgtta
//
