LOCUS       sequence-412700-        6365 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pLv1-pUC18R6KT-miniTn7-Km-mCherry-sYFP2, KmR, ApR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6365)
  AUTHORS   Jorrin B, Haskett TL, Knights HE, Martyn A, Underwood TJ, Dolliver 
            J, Ledermann R, Poole PS
  TITLE     Stable, fluorescent markers for tracking synthetic communities and 
            assembly dynamics.
  JOURNAL   Microbiome. 2024 May 7;12(1):81. doi: 10.1186/s40168-024-01792-2.
  PUBMED    38715147
REFERENCE   2  (bases 1 to 6365)
  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: 
            "Microbiome."; date: "2024-05-7"; volume: "12(1):81. doi"; pages: " 
            10.1186/s40168-024-01792-2"
COMMENT     Sequence Label: pTn7-SCOUT22.RY
FEATURES             Location/Qualifiers
     source          1..6365
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            1..109
                     /note="incP origin of transfer"
     rep_origin      160..516
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(join(925..1716,1717..1785))
                     /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     1548..1567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1786..1890)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1958..1976
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     2090..2109
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2523..2539
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2524..2540
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      2568..2662
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      2765..2851
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    2897..2944
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(2999..3814)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3806..3825
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    3955..4002
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        4079..4113
                     /label=BBa_J23104
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.72"
     RBS             4126..4137
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4145..4855
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4290..4308)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     RBS             5662..5673
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(5686..5720)
                     /label=BBa_J23104
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.72"
     terminator      5762..5856
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     mobile_element  5897..6062
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gggcaggata ggtgaagtag gcccacccgc gagcgggtgt tccttcttca ctgtccctta
       61 ttcgcacctg gcggtgctca acgggaatcc tgctctgcga ggctggccga taagctctga
      121 taacgcagga aagaacatgg gcagttcaac ctgttgatag tacgtactaa gctctcatgt
      181 ttcacgtact aagctctcaa gtttaacgta ctaagctctc atgtttaacg aactaaaccc
      241 tcatggctaa cgtactaagc tctcatggct aacgtactaa gctctcatgt ttcacgtact
      301 aagctctcat gtttgaacaa taaaattaat ataaatcagc aacttaaata gcctctaagg
      361 ttttaagttt tataagaaaa aaaagaatat ataaggcttt taaagctttt aaggtttaac
      421 ggttgtggac aacaagccag ggatgtaacg cactgagaag cccttagagc ctctcaaagc
      481 aattttgagt gacacaggaa cacttaacgg ctgacaaccg gctggtaaca ggattagcag
      541 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
      601 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
      661 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
      721 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
      781 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
      841 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
      901 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
      961 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1021 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgggacc cacgctcacc
     1081 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     1141 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     1201 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     1261 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     1321 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     1381 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     1441 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     1501 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     1561 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     1621 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     1681 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     1741 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     1801 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     1861 ttagaaaaat aaacaaatag gggttccgcg tacatttccc cgaaaagtgc cacctgacgt
     1921 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     1981 tcgtctaagc agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     2041 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     2101 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     2161 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     2221 gtgcgggcct cttcgctatt acgccagcta gaggaccagc cgcgtaacct ggcaaaatcg
     2281 gttacggttg agtaataaat ggatgccctg cgtaagcagg tgtgggcgga caataaagtc
     2341 ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt
     2401 tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct
     2461 aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggg
     2521 gttcgaggtc gacggtatcg ataagctagc ttaattagct gagcttggac tcctgttgat
     2581 agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg
     2641 ggcgtttttt attggtgaga atccaagcta gactgcgatg agtggcaggg cggggcgtaa
     2701 tttttttaag gcagttattg gtgcccttaa acgcctgggg taatgactct ctagcttgag
     2761 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     2821 gtcggtgaac gctctcctga gtaggacaaa tccgccgcta ggagcttgcg gcccggacga
     2881 tcgaattggg gatcttgaag tacctattcc gaagttccta ttctctagaa agtataggaa
     2941 cttcagagcg cttttgaagc tgatgtgctt aaaaacttac tcttagttca attaattatt
     3001 agaaaaattc atccagcatc agatgaaatt gcagtttgtt catatccgga ttatcaatgc
     3061 catatttctg aaacagacgt ttttgcaggc tcgggctaaa ttcgcccagg cagttccaca
     3121 gaatggccag atcctgataa cgatccgcaa tgcccacacg gcccacatca atgcagccaa
     3181 tcagtttgcc ttcatcgaaa atcaggttat ccaggctaaa atcgccgtgg gtcaccacgc
     3241 tatccgggct aaacggcagc agtttatgca tttctttcca cacctgttcc accggccagc
     3301 cgttacgttc atcatcaaaa tcgctcgcat ccaccaggcc gttgttcata cggctctgcg
     3361 cctgggccag acgaaacaca cgatcgctgt taaacgggca gttgcacacc ggaatgctat
     3421 gcagacgacg cagaaacacg gccagcgcat ccacaatgtt ttcgccgcta tccggatatt
     3481 cttccagcac ctgaaacgcg gttttgcccg gaatcgcggt ggtcagcagc cacgcatcat
     3541 ccggggtgcg aataaaatgt ttaatggtcg gcagcggcat aaattcggtc agccagttca
     3601 gacgcaccat ttcatcggtc acatcgttcg ccacgctgcc tttgccatgt ttcagaaaca
     3661 gttccggcgc atccggtttg ccatacagac gataaatggt cgcgccgctc tgacccacgt
     3721 tatcacgcgc ccatttatag ccatacagat ccgcatccat gttgctgttc agacgcggac
     3781 ggctacagct cgtttcacgc tgaatatggc tcataacacc ccttgtatta ctgtttatgt
     3841 aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga
     3901 gattttgaga cacaattgct ccattgctgt tgacaaaggg aatcagggga tcttgaagtt
     3961 cctattccga agttcctatt ctctagaaag tataggaact tcagagcgct tttgaagcta
     4021 attcgagatc atgcataatg ccgaattcgg atccggagtg ccgaattcgg atccggagtt
     4081 gacagctagc tcagtcctag gtattgtgct agctactcta gaattaaaga ggagaaatta
     4141 accaatggtg agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt
     4201 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
     4261 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
     4321 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
     4381 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
     4441 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
     4501 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     4561 cgtaatgcag aagaaaacca tgggctggga ggcctcctcc gagcggatgt accccgagga
     4621 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
     4681 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
     4741 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     4801 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtaagcttc
     4861 tcggtaccaa attccagaaa agaggcctcc cgaaaggggg gccttttttc tggaatttgg
     4921 taccgagaag cttattattt atacagctca tccataccca gggtaatacc ggcagcggtc
     4981 acaaactcca gcagcaccat gtggtcgcgc ttctcgttcg gatctttgct cagtttgctt
     5041 tggtaagaca gataatggtt atccggcaac aggaccggac cgtcaccaat cggggtgttt
     5101 tgctggtaat gatccgccag ttgaacacca ccgtcctcaa tgttgtgacg gattttgaaa
     5161 ttcgctttga tgccattttt ctgtttatct gcggtgatgt aaacgttgtg ggaattgtaa
     5221 ttgtactcca gcttgtgacc caggatattg ccatcctctt tgaaatcaat acccttcagc
     5281 tcaatgcgat tcaccagggt gtcgccttcg aatttgactt cggcacgcgt cttgtagtta
     5341 ccgtcgtctt tgaaaaagat cgtacgctcc tgcacatagc cttctggcat cgcgctctta
     5401 aagaaatcgt gttgtttcat gtggtccgga taacgcgcga aacactgaac gccataaccc
     5461 aaggtcgtga ccagggtagg ccacgggacc ggcagcttac ccgtcgtgca aatcagcttc
     5521 agggtcagct tgccgtaagt tgcgtcacct tcaccctcac cgctcacgct aaacttatga
     5581 ccgttaacgt cgccgtctaa ttcaactaag atcggtacta cgcctgtaaa aagttcttcg
     5641 cccttgctaa ccattggtta atttctcctc tttaattcta gagtagctag cacaatacct
     5701 aggactgagc tagctgtcaa ctccactata gaccgcagcg acggtctagg gaccgctctt
     5761 ggactcctgt tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg
     5821 ctcggttgcc gccgggcgtt ttttattggt gagaatccag gcaatcaatc gtgctttcgc
     5881 gaaggccttg caggccaacc agataagtga aatctagttc caaactattt tgtcattttt
     5941 aattttcgta ttagcttacg acgctacacc cagttcccat ctattttgtc actcttccct
     6001 aaataatcct taaaaactcc atttccaccc ctcccagttc ccaactattt tgtccgccca
     6061 cagcggggca tttttcttcc tgttatgttt gggcgctgca ttaatgaatc ggccaacgcg
     6121 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gacccgctgc
     6181 gctcggtcgt tcggctgcgg cgagcggtat caggggattc cttaaggtat actttccgct
     6241 gcataaccct gcttcggggt cattatagcg attttttcgg tatatccatc ctttttcgca
     6301 cgatatacag gattttgcca aagggttcgt gtagactttc cttggtgtat ccaacggcgt
     6361 cagcc
//
