LOCUS       sequence-366472-        6594 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 6594)
  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 6594)
  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-Tn5-147
FEATURES             Location/Qualifiers
     source          1..6594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            152..261
                     /note="incP origin of transfer"
     primer_bind     815..831
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     815..831
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        join(952..957,958..974,975..980)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1169..1702
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     CDS             join(2064..2066,2067..2579)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIE"
     primer_bind     complement(2106..2127)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2367..2386)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2714..2735
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      2764..2858
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2902..3561
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2968..2987)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3626..3712
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(5456..5474)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5542..5646
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5647..5715,5716..6507)
                     /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(5865..5884)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 attggggatc aattccgttt tccgctgcat aaccctgctt cggggtcatt atagcgattt
       61 tttcggtata tccatccttt ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag
      121 actttccttg gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta ggcccacccg
      181 cgagcgggtg ttccttcttc actgtccctt attcgcacct ggcggtgctc aacgggaatc
      241 ctgctctgcg aggctggccg gctaccgccg gcgtaacaga tgagggcaag cggatggctg
      301 atgaaaccaa gccaaccagg aagggcagcc cacctatcac ggaattgatc cggccacgat
      361 gcgtccggcg tagaggatct gaagatcagc agttcaacct gttgatagta cgtactaagc
      421 tctcatgttt cacgtactaa gctctcatgt ttaacgtact aagctctcat gtttaacgaa
      481 ctaaaccctc atggctaacg tactaagctc tcatgtttca cgtactaagc tctcatgttt
      541 gaacaataaa attaatataa atcagcaact taaatagcct ctaaggtttt aagttttata
      601 agaaaaaaaa gaatatataa ggcttttaaa gcttttaagg tttaacggtt gtggacaaca
      661 agccagggat gtaacgcact gagaagccct tagagcctct caaagcaatt ttgagtgaca
      721 caggaacact taacggctga catgggaatt aattctgact cttatacaca agtgcggccc
      781 tcgaattgac gcgctccacc gcggtggcgg ccgctctaga actagtggat cccccgggct
      841 gcaggaattc gatatccata aactgccagg aattggggat cggattcgag ctttagtacc
      901 tagatttaga tgtctaaaaa gcggggtacc gaggacgcgt caattctcga attgacataa
      961 gcctgttcgg ttcgtaaact gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga
     1021 accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac
     1081 tgtttttttg tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt
     1141 cgatgtttga tgttatggag cagcaacgat gttacgcagc agcaacgatg ttacgcagca
     1201 gggcagtcgc cctaaaacaa agttaggtgg ctcaagtatg ggcatcattc gcacatgtag
     1261 gctcggccct gaccaagtca aatccatgcg ggctgctctt gatcttttcg gtcgtgagtt
     1321 cggagacgta gccacctact cccaacatca gccggactcc gattacctcg ggaacttgct
     1381 ccgtagtaag acattcatcg cgcttgctgc cttcgaccaa gaagcggttg ttggcgctct
     1441 cgcggcttac gttctgccca agtttgagca gccgcgtagt gagatctata tctatgatct
     1501 cgcagtctcc ggcgagcacc ggaggcaggg cattgccacc gcgctcatca atctcctcaa
     1561 gcatgaggcc aacgcgcttg gtgcttatgt gatctacgtg caagcagatt acggtgacga
     1621 tcccgcagtg gctctctata caaagttggg catacgggaa gaagtgatgc actttgatat
     1681 cgacccaagt accgccacct aaagctcgca cgctgccgca agcactcagg gcgcaagggc
     1741 tgctaaagga agcggaacac gtagaaagcc agtccgcaga aacggtgctg accccggatg
     1801 aatgtcagct actgggctat ctggacaagg gaaaacgcaa gcgcaaagag aaagcaggta
     1861 gcttgcagtg ggcttacatg gcgatagcta gactgggcgg ttttatggac agcaagcgaa
     1921 ccggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa agtaaactgg
     1981 atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga tcaagagaca
     2041 ggatactagt ggaggaagaa aaaatgagca agggcgagga gctgttcacc ggggtggtgc
     2101 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     2161 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     2221 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcgccc
     2281 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     2341 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     2401 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     2461 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     2521 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg
     2581 acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2641 tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg
     2701 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggtaa ttagctgagc
     2761 ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa
     2821 cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagctagctt ggcgagattt
     2881 tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata
     2941 tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat
     3001 aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac
     3061 aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattt
     3121 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
     3181 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
     3241 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
     3301 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc
     3361 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
     3421 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
     3481 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
     3541 gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggg
     3601 gtaatgactc tctagcttga ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     3661 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccgct
     3721 aggagcttgc ggccgccagg atatcgaccc aagtaccgcc acctaacaat tcgttcaagc
     3781 cgagatcggc ttcccggccg acgcgtcaat tcgagcgcgt caattcgagg gccgcggcct
     3841 aggcggccag atctgatcaa gagacagtct agctagaatt aattcgagtc gaccgatccc
     3901 gtacacaagt agcgtcctga acggaacctt tcccgttttc caggatctga tcttccatgt
     3961 gacctcctaa catggtaacg ttcatgataa cttctgctct tcatcgtgcg gccgactggg
     4021 ctaaatctgt gttctcttcg gcggcgctgg gtgatcctcg ccgtactgcc cgcttggtta
     4081 acgtcgccgc ccaattggca aaatattctg gtaaatcaat aaccatctca tcagagggta
     4141 gtgaagccat gcaggaaggc gcttaccgat ttatccgcaa tcccaacgtt tctgccgagg
     4201 cgatcagaaa ggctggcgcc atgcaaacag tcaagttggc tcaggagttt cccgaactgc
     4261 tggccattga ggacaccacc tctttgagtt atcgccacca ggtcgccgaa gagcttggca
     4321 agctgggctc tattcaggat aaatcccgcg gatggtgggt tcactccgtt ctcttgctcg
     4381 aggccaccac attccgcacc gtaggattac tgcatcagga gtggtggatg cgcccggatg
     4441 accctgccga tgcggatgaa aaggagagtg gcaaatggct ggcagccgcc gcaactagcc
     4501 ggttacgcat gggcagcatg atgagcaacg tgattgcggt ctgtgaccgc gaagccgata
     4561 ttcatgctta tctgcaggac aaactggcgc ataacgagcg cttcgtggtg cgctccaagc
     4621 acccacgcaa ggacgtagag tctgggttgt atctgtacga ccatctgaag aaccaaccgg
     4681 agttgggtgg ctatcagatc agcattccgc aaaagggcgt ggtggataaa cgcggtaaac
     4741 gtaaaaatcg accagcccgc aaggcgagct tgagcctgcg cagtgggcgc atcacgctaa
     4801 aacaggggaa tatcacgctc aacgcggtgc tggccgagga gattaacccg cccaagggtg
     4861 agaccccgtt gaaatggttg ttgctgacca gcgaaccggt cgagtcgcta gcccaagcct
     4921 tgcgcgtcat cgacatttat acccatcgct ggcggatcga ggagttccat aaggcatgga
     4981 aaaccggagc aggagccgag aggcaacgca tggaggagcc ggataatctg gagcggatgg
     5041 tctcgatcct ctcgtttgtt gcggtcaggc tgttacagct cagagaaagc ttcacgctgc
     5101 cgcaagcact cagggcgcaa gggctgctaa aggaagcgga acacgtagaa agccagtccg
     5161 cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac aagggaaaac
     5221 gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata gctagactgg
     5281 gcggttttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc tgggaaggtt
     5341 gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg atggcgcagg
     5401 ggatcaagat ctgatcggtc gatcgaatta attcccaatt cttgaagacg aaagggcctc
     5461 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     5521 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5581 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5641 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5701 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5761 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5821 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5881 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5941 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     6001 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     6061 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     6121 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6181 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     6241 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     6301 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6361 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6421 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6481 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6541 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatccg ggga
//
