LOCUS       sequence-226854-        5766 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  miniTn5 plasmid to deliver constitutively expressed fluorescent 
            protein genes in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5766)
  AUTHORS   Schlechter RO, Jun H, Bernach M, Oso S, Boyd E, Munoz-Lintz DA, 
            Dobson RCJ, Remus DM, Remus-Emsermann MNP
  TITLE     Chromatic Bacteria - A Broad Host-Range Plasmid and Chromosomal 
            Insertion Toolbox for Fluorescent Protein Expression in Bacteria.
  JOURNAL   Front Microbiol. 2018 Dec 12;9:3052. doi: 10.3389/fmicb.2018.03052. 
            eCollection 2018.
  PUBMED    30631309
REFERENCE   2  (bases 1 to 5766)
  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: "Front 
            Microbiol."; date: "2018-12-12"; volume: "9:3052. doi"; pages: " 
            10.3389/fmicb.2018.03052. eCollection 2018"
COMMENT     Sequence Label: pMRE-Tn5-135
FEATURES             Location/Qualifiers
     source          1..5766
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     34..53
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      349..443
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             487..1146
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(553..572)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1211..1297
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3038..3056)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3124..3228
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3229..3297,3298..4089)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3447..3466)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            4328..4437
                     /note="incP origin of transfer"
     primer_bind     4991..5007
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4991..5007
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             join(5399..5766,1..328)
                     /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(5532..5550)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
ORIGIN
        1 gaagctccgc ggcaccaact tccctcctga cggccccgta atgcagaaga agacaatggg
       61 ctgggaagcg tccaccgagc ggttgtaccc cgaggacggc gtgctgaagg gcgacattaa
      121 gatggccctg cgcctgaagg acggcggccg ctacctggcg gacttcaaga ccacctacaa
      181 ggccaagaag cccgtgcaga tgcccggcgc ctacaacgtc gaccgcaagt tggacatcac
      241 ctcccacaac gaggactaca ccgtggtgga acagtacgaa cgctccgagg gccgccactc
      301 caccggcggc atggacgagc tgtacaagta agtaattagc tgagcttgga ctcctgttga
      361 tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc
      421 gggcgttttt tattggtgag aatccaagct agcttggcga gattttcagg agctaaggaa
      481 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
      541 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
      601 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
      661 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa
      721 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
      781 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
      841 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
      901 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
      961 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     1021 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     1081 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg
     1141 gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc ctggggtaat gactctctag
     1201 cttgaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     1261 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgctaggag cttgcggccg
     1321 ccagatcgac ccaagtaccg ccacctaaca attcgttcaa gccgagatcg gcttcccggc
     1381 cgacgcgtca attcgagcgc gtcaattcga gggccgcggc ctaggcggcc agatctgatc
     1441 aagagacagt ctagctagaa ttaattcgag tcgaccgatc ccgtacacaa gtagcgtcct
     1501 gaacggaacc tttcccgttt tccaggatct gatcttccat gtgacctcct aacatggtaa
     1561 cgttcatgat aacttctgct cttcatcgtg cggccgactg ggctaaatct gtgttctctt
     1621 cggcggcgct gggtgatcct cgccgtactg cccgcttggt taacgtcgcc gcccaattgg
     1681 caaaatattc tggtaaatca ataaccatct catcagaggg tagtgaagcc atgcaggaag
     1741 gcgcttaccg atttatccgc aatcccaacg tttctgccga ggcgatcaga aaggctggcg
     1801 ccatgcaaac agtcaagttg gctcaggagt ttcccgaact gctggccatt gaggacacca
     1861 cctctttgag ttatcgccac caggtcgccg aagagcttgg caagctgggc tctattcagg
     1921 ataaatcccg cggatggtgg gttcactccg ttctcttgct cgaggccacc acattccgca
     1981 ccgtaggatt actgcatcag gagtggtgga tgcgcccgga tgaccctgcc gatgcggatg
     2041 aaaaggagag tggcaaatgg ctggcagccg ccgcaactag ccggttacgc atgggcagca
     2101 tgatgagcaa cgtgattgcg gtctgtgacc gcgaagccga tattcatgct tatctgcagg
     2161 acaaactggc gcataacgag cgcttcgtgg tgcgctccaa gcacccacgc aaggacgtag
     2221 agtctgggtt gtatctgtac gaccatctga agaaccaacc ggagttgggt ggctatcaga
     2281 tcagcattcc gcaaaagggc gtggtggata aacgcggtaa acgtaaaaat cgaccagccc
     2341 gcaaggcgag cttgagcctg cgcagtgggc gcatcacgct aaaacagggg aatatcacgc
     2401 tcaacgcggt gctggccgag gagattaacc cgcccaaggg tgagaccccg ttgaaatggt
     2461 tgttgctgac cagcgaaccg gtcgagtcgc tagcccaagc cttgcgcgtc atcgacattt
     2521 atacccatcg ctggcggatc gaggagttcc ataaggcatg gaaaaccgga gcaggagccg
     2581 agaggcaacg catggaggag ccggataatc tggagcggat ggtctcgatc ctctcgtttg
     2641 ttgcggtcag gctgttacag ctcagagaaa gcttcacgct gccgcaagca ctcagggcgc
     2701 aagggctgct aaaggaagcg gaacacgtag aaagccagtc cgcagaaacg gtgctgaccc
     2761 cggatgaatg tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagagaaag
     2821 caggtagctt gcagtgggct tacatggcga tagctagact gggcggtttt atggacagca
     2881 agcgaaccgg aattgccagc tggggcgccc tctgggaagg ttgggaagcc ctgcaaagta
     2941 aactggatgg ctttcttgcc gccaaggatc tgatggcgca ggggatcaag atctgatcgg
     3001 tcgatcgaat taattcccaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     3061 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     3121 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3181 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     3241 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3301 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     3361 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     3421 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     3481 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     3541 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3601 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     3661 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3721 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     3781 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3841 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3901 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3961 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4021 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4081 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     4141 ttttaattta aaaggatcta ggtgaagatc cggggaattg gggatcaatt ccgttttccg
     4201 ctgcataacc ctgcttcggg gtcattatag cgattttttc ggtatatcca tcctttttcg
     4261 cacgatatac aggattttgc caaagggttc gtgtagactt tccttggtgt atccaacggc
     4321 gtcagccggg caggataggt gaagtaggcc cacccgcgag cgggtgttcc ttcttcactg
     4381 tcccttattc gcacctggcg gtgctcaacg ggaatcctgc tctgcgaggc tggccggcta
     4441 ccgccggcgt aacagatgag ggcaagcgga tggctgatga aaccaagcca accaggaagg
     4501 gcagcccacc tatcacggaa ttgatccggc cacgatgcgt ccggcgtaga ggatctgaag
     4561 atcagcagtt caacctgttg atagtacgta ctaagctctc atgtttcacg tactaagctc
     4621 tcatgtttaa cgtactaagc tctcatgttt aacgaactaa accctcatgg ctaacgtact
     4681 aagctctcat gtttcacgta ctaagctctc atgtttgaac aataaaatta atataaatca
     4741 gcaacttaaa tagcctctaa ggttttaagt tttataagaa aaaaaagaat atataaggct
     4801 tttaaagctt ttaaggttta acggttgtgg acaacaagcc agggatgtaa cgcactgaga
     4861 agcccttaga gcctctcaaa gcaattttga gtgacacagg aacacttaac ggctgacatg
     4921 ggaattaatt ctgactctta tacacaagtg cggccctcga attgacgcgc tccaccgcgg
     4981 tggcggccgc tctagaacta gtggatcccc cctgccagga attggggatc ggattcgagc
     5041 tcgcacgctg ccgcaagcac tcagggcgca agggctgcta aaggaagcgg aacacgtaga
     5101 aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga
     5161 caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat
     5221 agctagactg ggcggtctta tggacagcaa gcgaaccgga attgccagct ggggcgccct
     5281 ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct
     5341 gatggcgcag gggatcaaga tctgatcaag agacaggata ctagtggagg aagaaaaaat
     5401 ggtgagcaag ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc acatggaggg
     5461 ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     5521 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct cctgggacat
     5581 cctgtcccct cagttcatgt acggctccag ggccttcatc aagcaccccg ccgacatccc
     5641 cgactactat aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     5701 ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca ccctgatcta
     5761 caaggt
//
