LOCUS       sequence-394926-        6656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pLv1-pUC18R6KT-miniTn7-Tc-sfGFP, TcR, ApR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6656)
  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 6656)
  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-SCOUT13.G
FEATURES             Location/Qualifiers
     source          1..6656
                     /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(2988..4187)
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     CDS             4293..4943
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     protein_bind    4983..5030
                     /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)."
     terminator      5090..5176
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        5190..5224
                     /label=BBa_J23104
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.72"
     RBS             5237..5248
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5256..5969
                     /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      6053..6147
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     mobile_element  6188..6353
                     /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 gcgatcggct
     3001 cgttgccctg cgccgctcca aagcccgcga cgcagcgccg gcaggcagag caagtagagg
     3061 gcagcgcctg caatccatgc ccacccgttc cacgttgtta tagaagccgc atagatcgcc
     3121 gtgaagagga ggggtccgac gatcgaggtc aggctggtga gcgccgccag tgagccttgc
     3181 agctgcccct gacgttcctc atccacctgc ctggacaaca ttgcttgcag cgccggcatt
     3241 ccgatgccac ccgaagcaag caggaccatg atcgggaacg ccatccatcc ccgtgtcgcg
     3301 aaggcaagca ggatgtagcc tgtgccgtcg gcaatcattc cgagcatgag tgcccgcctt
     3361 tcgccgagcc gggcggctac agggccggtg atcattgcct gggcgagtga atgcagaatg
     3421 ccaaatgcgg caagcgaaat gccgatcgtg gtcgcgtccc agtgaaagcg atcctcgccg
     3481 aaaatgaccc aaagcgcggc cggcacctgt ccgacaagtt gcatgatgaa gaacaccgcc
     3541 atcagggcgg cgacgacggt catgccccgg gcccaccgga acgaagcgag cgggttgaga
     3601 gcctcccggc gtaacggccg gcgttcgcct ttgtgcgact ccggcaaaag gaaacagccc
     3661 gtcaggaaat tgaggccgtt caaggctgcc gcggcgaaga acggagcgtg gggggagaaa
     3721 ccgcccatca gcccaccgag cacaggtccc gcgaccatcc cgaacccgaa acaggcgctc
     3781 atgaagccga agtgccgcgc gcgctcatcg ccatcagtga tatcggcaat ataagcgccg
     3841 gctaccgccc cagtcgcccc ggtgatgccg gccacgatcc gcccgatata gagaacccaa
     3901 aggaaaggcg ctgtcgccat gatggcgtag tcgacagtgg cgccggccag cgacacgagc
     3961 aagattggcc gccgcccgaa acgatccgac agcgcgccca gcacaggtgc gcaggcaaat
     4021 tgcaccaacg catacagcgc cagcagaatg ccatagtggg cggtgacgtc gttcgagtga
     4081 accagatcgc gcaggaggcc cggcagcacc ggcataatca ggccgatgcc gacagcgtcg
     4141 agcgcgacag tgctcagaat tacgatcagg ggtatgttgg gtttcacgtc tggcctccgg
     4201 accagcctcc gctggtccga ttgaacgcgc ggattcttta tcactgataa gttggtggac
     4261 atattatgtt tatcagtgat aaagtgtcaa gcatgacaaa gttgcagccg aatacagtga
     4321 tccgtgccgc cctggacctg ttgaacgagg tcggcgtaga cggtctgacg acacgcaaac
     4381 tggcggaacg gttgggggtt cagcagccgg cgctttactg gcacttcagg aacaagcggg
     4441 cgctgctcga cgcactggcc gaagccatgc tggcggagaa tcatacgcat tcggtgccga
     4501 gagccgacga cgactggcgc tcatttctga tcgggaatgc ccgcagcttc aggcaggcgc
     4561 tgctcgccta ccgcgatggc gcgcgcatcc atgccggcac gcgaccgggc gcaccgcaga
     4621 tggaaacggc cgacgcgcag cttcgcttcc tctgcgaggc gggtttttcg gccggggacg
     4681 ccgtcaatgc gctgatgaca atcagctact tcactgttgg ggccgtgctt gaggagcagg
     4741 ccggcgacag cgatgccggc gagcgcggcg gcaccgttga acaggctccg ctctcgccgc
     4801 tgttgcgggc cgcgatagac gccttcgacg aagccggtcc ggacgcagcg ttcgagcagg
     4861 gactcgcggt gattgtcgat ggattggcga aaaggaggct cgttgtcagg aacgttgaag
     4921 gaccgagaaa gggtgacgat tgattgctcc attgctgttg acaaagggaa tcaggggatc
     4981 ttgaagttcc tattccgaag ttcctattct ctagaaagta taggaacttc agagcgcttt
     5041 tgaagctaat tcgagatcat gcataatgcc gaattcggat cccaggcatc aaataaaacg
     5101 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     5161 cctgagtagg acaaatccgc cgtccggagt tgacagctag ctcagtccta ggtattgtgc
     5221 tagctactct agaattaaag aggagaaatt aaccaatgcg taaaggcgaa gagctgttca
     5281 ctggtgtcgt ccctattctg gtggaactgg atggtgatgt caacggtcat aagttttccg
     5341 tgcgtggcga gggtgaaggt gacgcaacta atggtaaact gacgctgaag ttcatctgta
     5401 ctactggtaa actgccggta ccttggccga ctctggtaac gacgctgact tatggtgttc
     5461 agtgctttgc tcgttatccg gaccatatga agcagcatga cttcttcaag tccgccatgc
     5521 cggaaggcta tgtgcaggaa cgcacgattt cctttaagga tgacggcacg tacaaaacgc
     5581 gtgcggaagt gaaatttgaa ggcgataccc tggtaaaccg cattgagctg aaaggcattg
     5641 actttaaaga agatggcaat atcctgggcc ataagctgga atacaatttt aacagccaca
     5701 atgtttacat caccgccgat aaacaaaaaa atggcattaa agcgaatttt aaaattcgcc
     5761 acaacgttga ggatggcagc gtgcagctgg ctgatcacta ccagcaaaac actccaatcg
     5821 gtgatggtcc tgttctgctg ccagacaatc actatctgag cacgcaaagc gttctgtcta
     5881 aagatccgaa cgagaaacgc gatcatatgg ttctgctgga gttcgtaacc gcagcgggca
     5941 tcacgcatgg tatggatgaa ctgtacaaat gagcttctcg gtaccaaatt ccagaaaaga
     6001 ggcctcccga aaggggggcc ttttttcgtt ttggtcctcc ttggcgctct tggactcctg
     6061 ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc
     6121 cgccgggcgt tttttattgg tgagaatcca ggcaatcaat cgtgctttcg cgaaggcctt
     6181 gcaggccaac cagataagtg aaatctagtt ccaaactatt ttgtcatttt taattttcgt
     6241 attagcttac gacgctacac ccagttccca tctattttgt cactcttccc taaataatcc
     6301 ttaaaaactc catttccacc cctcccagtt cccaactatt ttgtccgccc acagcggggc
     6361 atttttcttc ctgttatgtt tgggcgctgc attaatgaat cggccaacgc gcggggagag
     6421 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgacccgctg cgctcggtcg
     6481 ttcggctgcg gcgagcggta tcaggggatt ccttaaggta tactttccgc tgcataaccc
     6541 tgcttcgggg tcattatagc gattttttcg gtatatccat cctttttcgc acgatataca
     6601 ggattttgcc aaagggttcg tgtagacttt ccttggtgta tccaacggcg tcagcc
//
