LOCUS       sequence-412701-        6337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pLv1-pUC18R6KT-miniTn7-Km-sYFP2-TagBFP, KmR, ApR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6337)
  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 6337)
  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.YB
FEATURES             Location/Qualifiers
     source          1..6337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1418..1429
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(1442..1476)
                     /label=BBa_J23104
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.72"
     terminator      1518..1612
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     mobile_element  1653..1818
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            2122..2230
                     /note="incP origin of transfer"
     CDS             complement(join(3024..3815,3816..3884))
                     /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     3647..3666
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3885..3989)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4057..4075
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     4189..4208
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4622..4638
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4623..4639
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      4667..4761
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      4864..4950
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    4996..5043
                     /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(5098..5913)
                     /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     5905..5924
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    6054..6101
                     /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        6178..6212
                     /label=BBa_J23104
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.72"
     RBS             6225..6236
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 gtgagggtga aggtgacgca acttacggca agctgaccct gaagctgatt tgcacgacgg
       61 gtaagctgcc ggtcccgtgg cctaccctgg tcacgacctt gggttatggc gttcagtgtt
      121 tcgcgcgtta tccggaccac atgaaacaac acgatttctt taagagcgcg atgccagaag
      181 gctatgtgca ggagcgtacg atctttttca aagacgacgg taactacaag acgcgtgccg
      241 aagtcaaatt cgaaggcgac accctggtga atcgcattga gctgaagggt attgatttca
      301 aagaggatgg caatatcctg ggtcacaagc tggagtacaa ttacaattcc cacaacgttt
      361 acatcaccgc agataaacag aaaaatggca tcaaagcgaa tttcaaaatc cgtcacaaca
      421 ttgaggacgg tggtgttcaa ctggcggatc attaccagca aaacaccccg attggtgacg
      481 gtccggtcct gttgccggat aaccattatc tgtcttacca aagcaaactg agcaaagatc
      541 cgaacgagaa gcgcgaccac atggtgctgc tggagtttgt gaccgctgcc ggtattaccc
      601 tgggtatgga tgagctgtat aaataataag cttctcggta ccaaattcca gaaaagaggc
      661 ccccctttcg ggaggcctct tttctggaat ttggtaccga gaagcttatt aattcagttt
      721 atgacccagc ttgctaggca agtcgcaata gcgcgcgact gcaacctcgt gctgctccac
      781 gtaagtctca ttattcgctt ctttaatgcg ttccagacgg tagtcgacat agtaaacacc
      841 cggcatcttc agatttttcg ccggtttctt gctgcggtac gtcgtcttga tatttgcgat
      901 caaatggctg ccaccaacca gcttcagtgc catgtcgtta cggccctcca ggccaccatc
      961 ggccggatac agggtttccg taaacgcctc ccaacccagg gttttcttct gcatcaccgg
     1021 accattgctc gtaaagttga cgccacgaat tttcacgtta tagatcagac agccatcttg
     1081 cagggaggta tcttgggtgg cggtcaacac accaccatct tcatacgtgg tgacacgctc
     1141 ccaggtgaaa ccttccggaa agctctgttt gaagaaatcc ggaatgccct gagtatggtt
     1201 aatgaacgtt ttggaaccgt acaggaagct ggtcgccaga atgtcaaaag cgaacggcag
     1261 tggaccgccc tccacaactt tgatacgcat ggtttgggtg ccttcgtacg gcttaccctc
     1321 accctcagac gtgcacttaa agtggtggtt gtcaacggtg ccttccatgt acagcttcat
     1381 gtgcatgttc tctttgatca gttcgctcat tggttaattt ctcctcttta attctagagt
     1441 agctagcaca atacctagga ctgagctagc tgtcaactcc actatagacc gcagcgacgg
     1501 tctagggacc gctcttggac tcctgttgat agatccagta atgacctcag aactccatct
     1561 ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaggcaa
     1621 tcaatcgtgc tttcgcgaag gccttgcagg ccaaccagat aagtgaaatc tagttccaaa
     1681 ctattttgtc atttttaatt ttcgtattag cttacgacgc tacacccagt tcccatctat
     1741 tttgtcactc ttccctaaat aatccttaaa aactccattt ccacccctcc cagttcccaa
     1801 ctattttgtc cgcccacagc ggggcatttt tcttcctgtt atgtttgggc gctgcattaa
     1861 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     1921 ctcactgacc cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagg ggattcctta
     1981 aggtatactt tccgctgcat aaccctgctt cggggtcatt atagcgattt tttcggtata
     2041 tccatccttt ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag actttccttg
     2101 gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta ggcccacccg cgagcgggtg
     2161 ttccttcttc actgtccctt attcgcacct ggcggtgctc aacgggaatc ctgctctgcg
     2221 aggctggccg ataagctctg ataacgcagg aaagaacatg ggcagttcaa cctgttgata
     2281 gtacgtacta agctctcatg tttcacgtac taagctctca tgtttaacga actaaaccct
     2341 catggctaac gtactaagct ctcatggcta acgtactaag ctctcatgtt tcacgtacta
     2401 agctctcatg tttgaacaat aaaattaata taaatcagca acttaaatag cctctaaggt
     2461 tttaagtttt ataagaaaaa aaagaatata taaggctttt aaagctttta aggtttaacg
     2521 gttgtggaca acaagccagg gatgtaacgc actgagaagc ccttagagcc tctcaaagca
     2581 attttgagtg acacaggaac acttaacggc tgacaaccgg ctggtaacag gattagcaga
     2641 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2701 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2761 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2821 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2881 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2941 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3001 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3061 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3121 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgggaccc acgctcaccg
     3181 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3241 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3301 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3361 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3421 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3481 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3541 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3601 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3661 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3721 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3781 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3841 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3901 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3961 tagaaaaata aacaaatagg ggttccgcgt acatttcccc gaaaagtgcc acctgacgtc
     4021 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     4081 cgtctaagca gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     4141 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     4201 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     4261 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     4321 tgcgggcctc ttcgctatta cgccagctag aggaccagcc gcgtaacctg gcaaaatcgg
     4381 ttacggttga gtaataaatg gatgccctgc gtaagcaggt gtgggcggac aataaagtct
     4441 taaactgaac aaaatagatc taaactatga caataaagtc ttaaactaga cagaatagtt
     4501 gtaaactgaa atcagtccag ttatgctgtg aaaaagcata ctggactttt gttatggcta
     4561 aagcaaactc ttcattttct gaagtgcaaa ttgcccgtcg tattaaagag gggcgtgggg
     4621 ttcgaggtcg acggtatcga taagctagct taattagctg agcttggact cctgttgata
     4681 gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg
     4741 gcgtttttta ttggtgagaa tccaagctag actgcgatga gtggcagggc ggggcgtaat
     4801 ttttttaagg cagttattgg tgcccttaaa cgcctggggt aatgactctc tagcttgagg
     4861 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     4921 tcggtgaacg ctctcctgag taggacaaat ccgccgctag gagcttgcgg cccggacgat
     4981 cgaattgggg atcttgaagt acctattccg aagttcctat tctctagaaa gtataggaac
     5041 ttcagagcgc ttttgaagct gatgtgctta aaaacttact cttagttcaa ttaattatta
     5101 gaaaaattca tccagcatca gatgaaattg cagtttgttc atatccggat tatcaatgcc
     5161 atatttctga aacagacgtt tttgcaggct cgggctaaat tcgcccaggc agttccacag
     5221 aatggccaga tcctgataac gatccgcaat gcccacacgg cccacatcaa tgcagccaat
     5281 cagtttgcct tcatcgaaaa tcaggttatc caggctaaaa tcgccgtggg tcaccacgct
     5341 atccgggcta aacggcagca gtttatgcat ttctttccac acctgttcca ccggccagcc
     5401 gttacgttca tcatcaaaat cgctcgcatc caccaggccg ttgttcatac ggctctgcgc
     5461 ctgggccaga cgaaacacac gatcgctgtt aaacgggcag ttgcacaccg gaatgctatg
     5521 cagacgacgc agaaacacgg ccagcgcatc cacaatgttt tcgccgctat ccggatattc
     5581 ttccagcacc tgaaacgcgg ttttgcccgg aatcgcggtg gtcagcagcc acgcatcatc
     5641 cggggtgcga ataaaatgtt taatggtcgg cagcggcata aattcggtca gccagttcag
     5701 acgcaccatt tcatcggtca catcgttcgc cacgctgcct ttgccatgtt tcagaaacag
     5761 ttccggcgca tccggtttgc catacagacg ataaatggtc gcgccgctct gacccacgtt
     5821 atcacgcgcc catttatagc catacagatc cgcatccatg ttgctgttca gacgcggacg
     5881 gctacagctc gtttcacgct gaatatggct cataacaccc cttgtattac tgtttatgta
     5941 agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag
     6001 attttgagac acaattgctc cattgctgtt gacaaaggga atcaggggat cttgaagttc
     6061 ctattccgaa gttcctattc tctagaaagt ataggaactt cagagcgctt ttgaagctaa
     6121 ttcgagatca tgcataatgc cgaattcgga tccggagtgc cgaattcgga tccggagttg
     6181 acagctagct cagtcctagg tattgtgcta gctactctag aattaaagag gagaaattaa
     6241 ccaatggtta gcaagggcga agaacttttt acaggcgtag taccgatctt agttgaatta
     6301 gacggcgacg ttaacggtca taagtttagc gtgagcg
//
