LOCUS       sequence-397600-        5396 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pLv1-pUC18R6KT-miniTn7-Gm-mCherry, GmR, ApR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5396)
  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 5396)
  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-SCOUT11.R
FEATURES             Location/Qualifiers
     source          1..5396
                     /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(3042..3575)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3764..3769,3770..3786,3787..3792))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    3832..3879
                     /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        3936..3970
                     /label=BBa_J23104
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.72"
     RBS             3983..3994
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4002..4712
                     /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(4147..4165)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      4793..4887
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     mobile_element  4928..5093
                     /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 tttaccgaac
     3001 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     3061 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     3121 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     3181 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc tccggagact
     3241 gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc
     3301 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     3361 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtcg
     3421 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     3481 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     3541 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatcaaaca
     3601 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcataggct gtacaaaaaa
     3661 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
     3721 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca
     3781 ggcttatgtc aattgctcca ttgctgttga caaagggaat caggggatct tgaagttcct
     3841 attccgaagt tcctattctc tagaaagtat aggaacttca gagcgctttt gaagctaatt
     3901 cgagatcatg cataatgccg aattcggatc cggagttgac agctagctca gtcctaggta
     3961 ttgtgctagc tactctagaa ttaaagagga gaaattaacc aatggtgagc aagggcgagg
     4021 aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg
     4081 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
     4141 agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt
     4201 cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact
     4261 acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     4321 gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg
     4381 tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag aaaaccatgg
     4441 gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca
     4501 agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca
     4561 aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca
     4621 cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact
     4681 ccaccggcgg catggacgag ctgtacaagt aagcttctcg gtaccaaatt ccagaaaaga
     4741 ggcctcccga aaggggggcc ttttttcgtt ttggtcctcc ttggcgctct tggactcctg
     4801 ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc
     4861 cgccgggcgt tttttattgg tgagaatcca ggcaatcaat cgtgctttcg cgaaggcctt
     4921 gcaggccaac cagataagtg aaatctagtt ccaaactatt ttgtcatttt taattttcgt
     4981 attagcttac gacgctacac ccagttccca tctattttgt cactcttccc taaataatcc
     5041 ttaaaaactc catttccacc cctcccagtt cccaactatt ttgtccgccc acagcggggc
     5101 atttttcttc ctgttatgtt tgggcgctgc attaatgaat cggccaacgc gcggggagag
     5161 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgacccgctg cgctcggtcg
     5221 ttcggctgcg gcgagcggta tcaggggatt ccttaaggta tactttccgc tgcataaccc
     5281 tgcttcgggg tcattatagc gattttttcg gtatatccat cctttttcgc acgatataca
     5341 ggattttgcc aaagggttcg tgtagacttt ccttggtgta tccaacggcg tcagcc
//
