LOCUS       sequence-227064-        6621 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 6621)
  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 6621)
  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-Tn5-144
FEATURES             Location/Qualifiers
     source          1..6621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             134..844
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     terminator      861..955
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             999..1658
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1065..1084)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1723..1809
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3553..3571)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3639..3743
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3744..3812,3813..4604)
                     /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(3962..3981)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            4843..4952
                     /note="incP origin of transfer"
     primer_bind     5506..5522
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5506..5522
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        join(5643..5648,5649..5665,5666..5671)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             5860..6393
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 cagctggggc gccctctggt aaggttggga agccctgcaa agtaaactgg atggctttct
       61 tgccgccaag gatctgatgg cgcaggggat caagatctga tcaagagaca ggatactagt
      121 ggaggaagaa aaaatggtga gcaagggcga ggagaataac atggccatca tcaaggagtt
      181 catgcgcttc aaggtgcgca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
      241 cgagggcgag ggccgcccct acgagggctt tcagaccgct aagctgaagg tgaccaaggg
      301 tggccccctg cccttcgcct gggacatcct gtcccctcat ttcacctacg gctccaaggc
      361 ctacgtgaag caccccgccg acatccccga ctacttcaag ctgtccttcc ccgagggctt
      421 caagtgggag cgcgtgatga actacgagga cggcggcgtg gtgaccgtga cccaggactc
      481 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
      541 cgacggcccc gtgatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
      601 ccccgaggac ggtgccctga agggcaagat caagatgagg ctgaagctga aggacggcgg
      661 ccactacacc tccgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
      721 cgcctacatc gtcgacatca agttggacat cacctcccac aacgaggact acaccatcgt
      781 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
      841 gtaataatta gctgagcttg gactcctgtt gatagatcca gtaatgacct cagaactcca
      901 tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag
      961 ctagcttggc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     1021 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     1081 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     1141 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     1201 atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg
     1261 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     1321 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     1381 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca
     1441 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     1501 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
     1561 cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat
     1621 tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt
     1681 gcccttaaac gcctggggta atgactctct agcttgaggc atcaaataaa acgaaaggct
     1741 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     1801 aggacaaatc cgccgctagg agcttgcggc cgccaggata tcgacccaag taccgccacc
     1861 taacaattcg ttcaagccga gatcggcttc ccggccgacg cgtcaattcg agcgcgtcaa
     1921 ttcgagggcc gcggcctagg cggccagatc tgatcaagag acagtctagc tagaattaat
     1981 tcgagtcgac cgatcccgta cacaagtagc gtcctgaacg gaacctttcc cgttttccag
     2041 gatctgatct tccatgtgac ctcctaacat ggtaacgttc atgataactt ctgctcttca
     2101 tcgtgcggcc gactgggcta aatctgtgtt ctcttcggcg gcgctgggtg atcctcgccg
     2161 tactgcccgc ttggttaacg tcgccgccca attggcaaaa tattctggta aatcaataac
     2221 catctcatca gagggtagtg aagccatgca ggaaggcgct taccgattta tccgcaatcc
     2281 caacgtttct gccgaggcga tcagaaaggc tggcgccatg caaacagtca agttggctca
     2341 ggagtttccc gaactgctgg ccattgagga caccacctct ttgagttatc gccaccaggt
     2401 cgccgaagag cttggcaagc tgggctctat tcaggataaa tcccgcggat ggtgggttca
     2461 ctccgttctc ttgctcgagg ccaccacatt ccgcaccgta ggattactgc atcaggagtg
     2521 gtggatgcgc ccggatgacc ctgccgatgc ggatgaaaag gagagtggca aatggctggc
     2581 agccgccgca actagccggt tacgcatggg cagcatgatg agcaacgtga ttgcggtctg
     2641 tgaccgcgaa gccgatattc atgcttatct gcaggacaaa ctggcgcata acgagcgctt
     2701 cgtggtgcgc tccaagcacc cacgcaagga cgtagagtct gggttgtatc tgtacgacca
     2761 tctgaagaac caaccggagt tgggtggcta tcagatcagc attccgcaaa agggcgtggt
     2821 ggataaacgc ggtaaacgta aaaatcgacc agcccgcaag gcgagcttga gcctgcgcag
     2881 tgggcgcatc acgctaaaac aggggaatat cacgctcaac gcggtgctgg ccgaggagat
     2941 taacccgccc aagggtgaga ccccgttgaa atggttgttg ctgaccagcg aaccggtcga
     3001 gtcgctagcc caagccttgc gcgtcatcga catttatacc catcgctggc ggatcgagga
     3061 gttccataag gcatggaaaa ccggagcagg agccgagagg caacgcatgg aggagccgga
     3121 taatctggag cggatggtct cgatcctctc gtttgttgcg gtcaggctgt tacagctcag
     3181 agaaagcttc acgctgccgc aagcactcag ggcgcaaggg ctgctaaagg aagcggaaca
     3241 cgtagaaagc cagtccgcag aaacggtgct gaccccggat gaatgtcagc tactgggcta
     3301 tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt gggcttacat
     3361 ggcgatagct agactgggcg gttttatgga cagcaagcga accggaattg ccagctgggg
     3421 cgccctctgg gaaggttggg aagccctgca aagtaaactg gatggctttc ttgccgccaa
     3481 ggatctgatg gcgcagggga tcaagatctg atcggtcgat cgaattaatt cccaattctt
     3541 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     3601 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3661 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3721 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3781 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3841 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3901 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3961 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4021 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4081 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4141 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4201 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4261 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4321 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4381 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4441 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4501 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4561 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4621 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4681 agatccgggg aattggggat caattccgtt ttccgctgca taaccctgct tcggggtcat
     4741 tatagcgatt ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag
     4801 ggttcgtgta gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt
     4861 aggcccaccc gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct
     4921 caacgggaat cctgctctgc gaggctggcc ggctaccgcc ggcgtaacag atgagggcaa
     4981 gcggatggct gatgaaacca agccaaccag gaagggcagc ccacctatca cggaattgat
     5041 ccggccacga tgcgtccggc gtagaggatc tgaagatcag cagttcaacc tgttgatagt
     5101 acgtactaag ctctcatgtt tcacgtacta agctctcatg tttaacgtac taagctctca
     5161 tgtttaacga actaaaccct catggctaac gtactaagct ctcatgtttc acgtactaag
     5221 ctctcatgtt tgaacaataa aattaatata aatcagcaac ttaaatagcc tctaaggttt
     5281 taagttttat aagaaaaaaa agaatatata aggcttttaa agcttttaag gtttaacggt
     5341 tgtggacaac aagccaggga tgtaacgcac tgagaagccc ttagagcctc tcaaagcaat
     5401 tttgagtgac acaggaacac ttaacggctg acatgggaat taattctgac tcttatacac
     5461 aagtgcggcc ctcgaattga cgcgctccac cgcggtggcg gccgctctag aactagtgga
     5521 tcccccgggc tgcaggaatt cgatatccat aaactgccag gaattgggga tcggattcga
     5581 gctttagtac ctagatttag atgtctaaaa agcggggtac cgaggacgcg tcaattctcg
     5641 aattgacata agcctgttcg gttcgtaaac tgtaatgcaa gtagcgtatg cgctcacgca
     5701 actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
     5761 cttgttatga ctgttttttt gtacagtcta tgcctcgggc atccaagcag caagcgcgtt
     5821 acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagcaacgat
     5881 gttacgcagc agggcagtcg ccctaaaaca aagttaggtg gctcaagtat gggcatcatt
     5941 cgcacatgta ggctcggccc tgaccaagtc aaatccatgc gggctgctct tgatcttttc
     6001 ggtcgtgagt tcggagacgt agccacctac tcccaacatc agccggactc cgattacctc
     6061 gggaacttgc tccgtagtaa gacattcatc gcgcttgctg ccttcgacca agaagcggtt
     6121 gttggcgctc tcgcggctta cgttctgccc aagtttgagc agccgcgtag tgagatctat
     6181 atctatgatc tcgcagtctc cggcgagcac cggaggcagg gcattgccac cgcgctcatc
     6241 aatctcctca agcatgaggc caacgcgctt ggtgcttatg tgatctacgt gcaagcagat
     6301 tacggtgacg atcccgcagt ggctctctat acaaagttgg gcatacggga agaagtgatg
     6361 cactttgata tcgacccaag taccgccacc taaagctcgc acgctgccgc aagcactcag
     6421 ggcgcaaggg ctgctaaagg aagcggaaca cgtagaaagc cagtccgcag aaacggtgct
     6481 gaccccggat gaatgtcagc tactgggcta tctggacaag ggaaaacgca agcgcaaaga
     6541 gaaagcaggt agcttgcagt gggcttacat ggcgatagct agactgggcg gttttatgga
     6601 cagcaagcga accggaattg c
//
