LOCUS       sequence-377519-        5446 bp    DNA     circular SYN 15-APR-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 5446)
  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 5446)
  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-137
FEATURES             Location/Qualifiers
     source          1..5446
                     /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..2495)
                     /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(1821..1842)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2429..2450
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      2512..2606
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2650..3309
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2716..2735)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3374..3460
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3836..3858
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3850..3867
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3851..3867
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     oriT            3955..4063
                     /note="incP origin of transfer"
     protein_bind    4210..4231
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4246..4251,4252..4269,4270..4276)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4284..4300
                     /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     4289..4311
                     /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 gcgtccgcgg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg aagttcatct
     1921 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc ggctacggcg
     1981 tggcctgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2041 tgcccgaagg ctacgtccag gagcgcacca tctctttcaa ggacgacggt acctacaaga
     2101 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     2161 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac ttcaacagcc
     2221 actacgtcta tatcacggcc gacaagcaga agaactgcat caaggctaac ttcaagatcc
     2281 gccacaacgt tgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     2341 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagccatcag tccaagctga
     2401 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     2461 ggattacaca tggcatggac gagctgtaca agtaataatt agctgagctt ggactcctgt
     2521 tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc
     2581 gccgggcgtt ttttattggt gagaatccaa gctagcttgg cgagattttc aggagctaag
     2641 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     2701 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     2761 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
     2821 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaatttcg tatggcaatg
     2881 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     2941 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     3001 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     3061 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
     3121 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     3181 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     3241 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     3301 ggggcgtaat ttttttaagg cagttattgg tgcccttaaa cgcctggggt aatgactctc
     3361 tagcttgagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     3421 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgctag gagcttgcgg
     3481 ccgccagccc gggcaattcg aggggtatcg ctcttgaagg gaactatgtt gaataataaa
     3541 aacgaatttt cacaaaaaaa tgtgtttttc tttgttagac cggggactta tcagccaacc
     3601 tgttatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3661 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3721 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3781 ccaacttact tctgacaacg atcgaggcga ttaagttggg taacgccagg gttttcccag
     3841 tcacgacgtt gtaaaacgac ggccagtgaa ttaattcttg aagacgaaag ggcctcgtga
     3901 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagagc ttatcggcca
     3961 gcctcgcaga gcaggattcc cgttgagcac cgccaggtgc gaataaggga cagtgaagaa
     4021 ggaacacccg ctcgcgggtg ggcctacttc acctatcctg cccggctgac gccgttggat
     4081 acaccaagga aagtctacac gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga
     4141 tggatatacc gaaaaaatcg ctataatgac cccgaagcag ggttatgcag cgggtgagcg
     4201 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     4261 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacacata
     4321 tggaaaaaaa ggaatttcgt gttttgataa aatactgttt tctgaaggga aaaaatacag
     4381 tggaagcaaa aacttggctt gataatgagt ttccggactc tgccccaggg aaatcaacaa
     4441 taattgattg gtatgcaaaa ttcaagcgtg gtgaaatgag cacggaggac ggtgaacgca
     4501 gtggacgccc gaaagaggtg gttaccgacg aaaacatcaa aaaaatccac aaaatgattt
     4561 tgaatgaccg taaaatgaag ttgatcgaga tagcagaggc cttaaagata tcaaaggaac
     4621 gtgttggtca tatcattcat caatatttgg atatgcggaa gctctgtgca aaatgggtgc
     4681 cgcgcgagct cacatttgac caaaaacaac aacgtgttga tgattctgag cggtgtttgc
     4741 agctgttaac tcgtaataca cccgagtttt tccgtcgata tgtgacaatg gatgaaacat
     4801 ggctccatca ctacactcct gagtccaatc gacagtcggc tgagtggaca gcgaccggtg
     4861 aaccgtctcc gaagcgtgga aagactcaaa agtccgctgg caaagtaatg gcctctgttt
     4921 tttgggatgc gcatggaata atttttatcg attatcttga gaagggaaaa accatcaaca
     4981 gtgactatta tatggcgtta ttggagcgtt tgaaggtcga aatcgcggca aaacggcccc
     5041 acatgaagaa gaaaaaagtg ttgttccacc aagacaacgc accgtgccac aagtcattga
     5101 gaacgatggc aaaaattcat gaattgggct tcgaattgct tccccacccg ccgtattctc
     5161 cagatctggc ccccagcgac tttttcttgt tctcagacct caaaaggatg ctcgcaggga
     5221 aaaaatttgg ctgcaatgaa gaggtgatcg ccgaaactga ggcctatttt gaggcaaaac
     5281 cgaaggagta ctaccaaaat ggtatcaaaa aattggaagg tcgttataat cgttgtatcg
     5341 ctcttgaagg gaactatgtt gaataataaa aacgaatttt cacaaaaaaa tgtgtttttc
     5401 tttgttagac cggatctgca gaattccagc acactggcgg ccgtta
//
