LOCUS       sequence-227065-        6610 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 6610)
  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 6610)
  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-Tn5-145
FEATURES             Location/Qualifiers
     source          1..6610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..696
                     /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(134..152)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     402..421
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      717..811
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             855..1514
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(921..940)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1579..1665
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3409..3427)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3495..3599
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3600..3668,3669..4460)
                     /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(3818..3837)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            4699..4808
                     /note="incP origin of transfer"
     primer_bind     5362..5378
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5362..5378
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        join(5499..5504,5505..5521,5522..5527)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             5716..6249
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 atggtgagca agggcgaggc agtgatcaag gagttcatgc ggttcaaggt gcacatggag
       61 ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
      121 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt ctcctgggac
      181 atcctgtccc ctcagttcat gtacggctcc agggccttca tcaagcaccc cgccgacatc
      241 cccgactact ataagcagtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
      301 gaggacggcg gcgccgtgac cgtgacccag gacacctccc tggaggacgg caccctgatc
      361 tacaaggtga agctccgcgg caccaacttc cctcctgacg gccccgtaat gcagaagaag
      421 acaatgggct gggaagcgtc caccgagcgg ttgtaccccg aggacggcgt gctgaagggc
      481 gacattaaga tggccctgcg cctgaaggac ggcggccgct acctggcgga cttcaagacc
      541 acctacaagg ccaagaagcc cgtgcagatg cccggcgcct acaacgtcga ccgcaagttg
      601 gacatcacct cccacaacga ggactacacc gtggtggaac agtacgaacg ctccgagggc
      661 cgccactcca ccggcggcat ggacgagctg tacaagtaag taattagctg agcttggact
      721 cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg
      781 ttgccgccgg gcgtttttta ttggtgagaa tccaagctag cttggcgaga ttttcaggag
      841 ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat
      901 ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga
      961 ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt
     1021 atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa tttcgtatgg
     1081 caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc
     1141 atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt
     1201 ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta
     1261 aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt
     1321 ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat
     1381 attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt
     1441 gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc
     1501 agggcggggc gtaatttttt taaggcagtt attggtgccc ttaaacgcct ggggtaatga
     1561 ctctctagct tgaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     1621 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gctaggagct
     1681 tgcggccgcc aggatatcga cccaagtacc gccacctaac aattcgttca agccgagatc
     1741 ggcttcccgg ccgacgcgtc aattcgagcg cgtcaattcg agggccgcgg cctaggcggc
     1801 cagatctgat caagagacag tctagctaga attaattcga gtcgaccgat cccgtacaca
     1861 agtagcgtcc tgaacggaac ctttcccgtt ttccaggatc tgatcttcca tgtgacctcc
     1921 taacatggta acgttcatga taacttctgc tcttcatcgt gcggccgact gggctaaatc
     1981 tgtgttctct tcggcggcgc tgggtgatcc tcgccgtact gcccgcttgg ttaacgtcgc
     2041 cgcccaattg gcaaaatatt ctggtaaatc aataaccatc tcatcagagg gtagtgaagc
     2101 catgcaggaa ggcgcttacc gatttatccg caatcccaac gtttctgccg aggcgatcag
     2161 aaaggctggc gccatgcaaa cagtcaagtt ggctcaggag tttcccgaac tgctggccat
     2221 tgaggacacc acctctttga gttatcgcca ccaggtcgcc gaagagcttg gcaagctggg
     2281 ctctattcag gataaatccc gcggatggtg ggttcactcc gttctcttgc tcgaggccac
     2341 cacattccgc accgtaggat tactgcatca ggagtggtgg atgcgcccgg atgaccctgc
     2401 cgatgcggat gaaaaggaga gtggcaaatg gctggcagcc gccgcaacta gccggttacg
     2461 catgggcagc atgatgagca acgtgattgc ggtctgtgac cgcgaagccg atattcatgc
     2521 ttatctgcag gacaaactgg cgcataacga gcgcttcgtg gtgcgctcca agcacccacg
     2581 caaggacgta gagtctgggt tgtatctgta cgaccatctg aagaaccaac cggagttggg
     2641 tggctatcag atcagcattc cgcaaaaggg cgtggtggat aaacgcggta aacgtaaaaa
     2701 tcgaccagcc cgcaaggcga gcttgagcct gcgcagtggg cgcatcacgc taaaacaggg
     2761 gaatatcacg ctcaacgcgg tgctggccga ggagattaac ccgcccaagg gtgagacccc
     2821 gttgaaatgg ttgttgctga ccagcgaacc ggtcgagtcg ctagcccaag ccttgcgcgt
     2881 catcgacatt tatacccatc gctggcggat cgaggagttc cataaggcat ggaaaaccgg
     2941 agcaggagcc gagaggcaac gcatggagga gccggataat ctggagcgga tggtctcgat
     3001 cctctcgttt gttgcggtca ggctgttaca gctcagagaa agcttcacgc tgccgcaagc
     3061 actcagggcg caagggctgc taaaggaagc ggaacacgta gaaagccagt ccgcagaaac
     3121 ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa aacgcaagcg
     3181 caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac tgggcggttt
     3241 tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctgggaag gttgggaagc
     3301 cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc aggggatcaa
     3361 gatctgatcg gtcgatcgaa ttaattccca attcttgaag acgaaagggc ctcgtgatac
     3421 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     3481 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3541 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3601 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3661 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3721 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3781 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3841 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3901 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3961 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4021 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4081 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4141 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4201 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4261 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4321 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4381 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4441 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4501 taaaacttca tttttaattt aaaaggatct aggtgaagat ccggggaatt ggggatcaat
     4561 tccgttttcc gctgcataac cctgcttcgg ggtcattata gcgatttttt cggtatatcc
     4621 atcctttttc gcacgatata caggattttg ccaaagggtt cgtgtagact ttccttggtg
     4681 tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc
     4741 cttcttcact gtcccttatt cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg
     4801 ctggccggct accgccggcg taacagatga gggcaagcgg atggctgatg aaaccaagcc
     4861 aaccaggaag ggcagcccac ctatcacgga attgatccgg ccacgatgcg tccggcgtag
     4921 aggatctgaa gatcagcagt tcaacctgtt gatagtacgt actaagctct catgtttcac
     4981 gtactaagct ctcatgttta acgtactaag ctctcatgtt taacgaacta aaccctcatg
     5041 gctaacgtac taagctctca tgtttcacgt actaagctct catgtttgaa caataaaatt
     5101 aatataaatc agcaacttaa atagcctcta aggttttaag ttttataaga aaaaaaagaa
     5161 tatataaggc ttttaaagct tttaaggttt aacggttgtg gacaacaagc cagggatgta
     5221 acgcactgag aagcccttag agcctctcaa agcaattttg agtgacacag gaacacttaa
     5281 cggctgacat gggaattaat tctgactctt atacacaagt gcggccctcg aattgacgcg
     5341 ctccaccgcg gtggcggccg ctctagaact agtggatccc ccgggctgca ggaattcgat
     5401 atccataaac tgccaggaat tggggatcgg attcgagctt tagtacctag atttagatgt
     5461 ctaaaaagcg gggtaccgag gacgcgtcaa ttctcgaatt gacataagcc tgttcggttc
     5521 gtaaactgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
     5581 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgtac
     5641 agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt
     5701 tatggagcag caacgatgtt acgcagcagc aacgatgtta cgcagcaggg cagtcgccct
     5761 aaaacaaagt taggtggctc aagtatgggc atcattcgca catgtaggct cggccctgac
     5821 caagtcaaat ccatgcgggc tgctcttgat cttttcggtc gtgagttcgg agacgtagcc
     5881 acctactccc aacatcagcc ggactccgat tacctcggga acttgctccg tagtaagaca
     5941 ttcatcgcgc ttgctgcctt cgaccaagaa gcggttgttg gcgctctcgc ggcttacgtt
     6001 ctgcccaagt ttgagcagcc gcgtagtgag atctatatct atgatctcgc agtctccggc
     6061 gagcaccgga ggcagggcat tgccaccgcg ctcatcaatc tcctcaagca tgaggccaac
     6121 gcgcttggtg cttatgtgat ctacgtgcaa gcagattacg gtgacgatcc cgcagtggct
     6181 ctctatacaa agttgggcat acgggaagaa gtgatgcact ttgatatcga cccaagtacc
     6241 gccacctaaa gctcgcacgc tgccgcaagc actcagggcg caagggctgc taaaggaagc
     6301 ggaacacgta gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact
     6361 gggctatctg gacaagggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc
     6421 ttacatggcg atagctagac tgggcggtct tatggacagc aagcgaaccg gaattgccag
     6481 ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc
     6541 cgccaaggat ctgatggcgc aggggatcaa gatctgatca agagacagga tactagtgga
     6601 ggaagaaaaa
//
