LOCUS       sequence-413655-        5519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pLv1-pUC18R6KT-miniTn7-aad9-sfGFP, SpR, StR, ApR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5519)
  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 5519)
  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-SCOUT14.G
FEATURES             Location/Qualifiers
     source          1..5519
                     /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)."
     protein_bind    3946..3993
                     /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        4050..4084
                     /label=BBa_J23104
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.72"
     RBS             4097..4108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4116..4829
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      4916..5010
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     mobile_element  5051..5216
                     /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 tcttagtcaa aatcggctcc
     3001 gtcggaattc ataataaaac aaaaaaattg aaaaaagtgt ttccaccatt ttttcaattt
     3061 ttttacagtt ttttcaggcg attattcaga taattaatgg tcagattcac attttcattg
     3121 gtccattcaa tattttcgcc cagatagctg cgcacggcca gcagaatgcg ttcgcggtgt
     3181 tccagcgggc tgctttcggc cacggcattg ccggcaatat ctttcggaat aattttgccg
     3241 gtatccatgg tcagaatcat gcggcacagg gtcagaatgc tattggtttc atcatcctga
     3301 taattatcaa tcagttcttc gctgctatcc ataatggcgc ggcgcacatc gctaaacgga
     3361 atatccggca gcagttcttc cagatcataa ttgccataaa tgcgtttatt tttgcgtttg
     3421 gcctgataca gcataatggt cagatcgcta ttcagttctt tctgcggaat atagccctgt
     3481 tcatacagtt cttgcagcca ttcgccataa ataaattcct gtttcggcgg atgattccac
     3541 ggcaccattt cctgctgaat aataatggtc agttcaatat agcgcagatt gcttttatcg
     3601 ccaatttttt tgctaatcgg gcgaattttc tgaatcagaa tttctttgct ctgatcggtc
     3661 agcggttcgc tcaccaccac cagaaaatcc agatcgctat tcggtttcag gccgctttcc
     3721 acgccgctgc caaacatata ggtgccaatc agattatttt tcagatgttt gcgcagaatt
     3781 tttttcactt tattaatctg ttcataggta ttcatataaa tcctcctcag gataactcaa
     3841 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     3901 gatcaattgc tccattgctg ttgacaaagg gaatcagggg atcttgaagt tcctattccg
     3961 aagttcctat tctctagaaa gtataggaac ttcagagcgc ttttgaagct aattcgagat
     4021 catgcataat gccgaattcg gatccggagt tgacagctag ctcagtccta ggtattgtgc
     4081 tagctactct agaattaaag aggagaaatt aaccaatgcg taaaggcgaa gagctgttca
     4141 ctggtgtcgt ccctattctg gtggaactgg atggtgatgt caacggtcat aagttttccg
     4201 tgcgtggcga gggtgaaggt gacgcaacta atggtaaact gacgctgaag ttcatctgta
     4261 ctactggtaa actgccggta ccttggccga ctctggtaac gacgctgact tatggtgttc
     4321 agtgctttgc tcgttatccg gaccatatga agcagcatga cttcttcaag tccgccatgc
     4381 cggaaggcta tgtgcaggaa cgcacgattt cctttaagga tgacggcacg tacaaaacgc
     4441 gtgcggaagt gaaatttgaa ggcgataccc tggtaaaccg cattgagctg aaaggcattg
     4501 actttaaaga agatggcaat atcctgggcc ataagctgga atacaatttt aacagccaca
     4561 atgtttacat caccgccgat aaacaaaaaa atggcattaa agcgaatttt aaaattcgcc
     4621 acaacgttga ggatggcagc gtgcagctgg ctgatcacta ccagcaaaac actccaatcg
     4681 gtgatggtcc tgttctgctg ccagacaatc actatctgag cacgcaaagc gttctgtcta
     4741 aagatccgaa cgagaaacgc gatcatatgg ttctgctgga gttcgtaacc gcagcgggca
     4801 tcacgcatgg tatggatgaa ctgtacaaat gagcttctcg gtaccaaatt ccagaaaaga
     4861 ggcctcccga aaggggggcc ttttttcgat aagctagctt aattagctga gcttggactc
     4921 ctgttgatag atccagtaat gacctcagaa ctccatctgg atttgttcag aacgctcggt
     4981 tgccgccggg cgttttttat tggtgagaat ccaggcaatc aatcgtgctt tcgcgaaggc
     5041 cttgcaggcc aaccagataa gtgaaatcta gttccaaact attttgtcat ttttaatttt
     5101 cgtattagct tacgacgcta cacccagttc ccatctattt tgtcactctt ccctaaataa
     5161 tccttaaaaa ctccatttcc acccctccca gttcccaact attttgtccg cccacagcgg
     5221 ggcatttttc ttcctgttat gtttgggcgc tgcattaatg aatcggccaa cgcgcgggga
     5281 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgacccg ctgcgctcgg
     5341 tcgttcggct gcggcgagcg gtatcagggg attccttaag gtatactttc cgctgcataa
     5401 ccctgcttcg gggtcattat agcgattttt tcggtatatc catccttttt cgcacgatat
     5461 acaggatttt gccaaagggt tcgtgtagac tttccttggt gtatccaacg gcgtcagcc
//
