LOCUS       sequence-446746-        7428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for inserting SnoopCatcher fused to mNeonGreen at the bacterial
            Tn7 site .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7428)
  AUTHORS   Ortiz de Ora L, Wiles ET, Zund M, Banuelos MS, Haro-Ramirez N, Suder
            DS, Ujagar N, Ayala-Angulo J, Trinh C, Knitter C, Gonen S, Nicholas 
            DA, Wiles TJ
  TITLE     Phollow reveals in situ phage transmission dynamics in the zebrafish
            gut microbiome at single-virion resolution.
  JOURNAL   Nat Microbiol. 2025 Apr 18. doi: 10.1038/s41564-025-01981-1.
  PUBMED    40251446
REFERENCE   2  (bases 1 to 7428)
  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: "Nat 
            Microbiol. 2025 Apr 18. doi: 10.1038/s41564-025-01981-1."
COMMENT     Sequence Label: pTn7xTS_SnoopCatcher:mNG
FEATURES             Location/Qualifiers
     source          1..7428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      638..860
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             908..1858
                     /codon_start=1
                     /gene="rep101 encoding an A56V mutation"
                     /product="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /label=Rep101(Ts)
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     CDS             complement(join(2517..3308,3309..3377))
                     /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     3140..3159
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3378..3482)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3550..3568
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3606..3628)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3728..3747
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4161..4177
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4162..4178
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      4206..4300
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      4403..4489
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    4550..4597
                     /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(4727..5260)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5449..5454,5455..5471,5472..5477))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    5518..5565
                     /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        join(5620..5625,5626..5641,5642..5648)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     RBS             5672..5694
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5706..6041
                     /label=SnoopCatcher
     CDS             6057..6767
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      6312..6321
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     mobile_element  6909..7074
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            join(7378..7428,1..58)
                     /note="incP origin of transfer"
ORIGIN
        1 tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag gctggccgat
       61 aagctctgat aacgcaggaa agaacatggg cagttcaacc tgttgatagt acatgaccaa
      121 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgttgatg ataccgctgc
      181 cttactgggt gcattagcca gtctgaatga cctgtcacgg gataatccga agtggtcaga
      241 ctggaaaatc agagggcagg aactgctgaa cagcaaaaag tcagatagca ccacatagca
      301 gacccgccat aaaacgccct gagaagcccg tgacgggctt ttcttgtatt atgggtagtt
      361 tccttgcatg aatccataaa aggcgcctgt agtgccattt acccccattc actgccagag
      421 ccgtgagcgc agcgaactga atgtcacgaa aaagacagcg actcaggtgc ctgatggtcg
      481 gagacaaaag gaatattcag cgatttgccc gattgcggcc gcaaccgagc ttgcgagggt
      541 gctacttaag cctttagggt tttaaggtct gttttgtaga ggagcaaaca gcgtttgcga
      601 catccttttg taatactgcg gaactgacta aagtagtgag ttatacacag ggctgggatc
      661 tattcttttt atcttttttt attctttctt tattctataa attataacca cttgaatata
      721 aacaaaaaaa acacacaaag gtctagcgga atttacagag ggtctagcag aatttacaag
      781 ttttccagca aaggtctagc agaatttaca gatacccaca actcaaagga aaaggactag
      841 taattatcat tgactagccc atctcaattg gtatagtgat taaaatcacc tagaccaatt
      901 gagatgtatg tctgaattag ttgttttcaa agcaaatgaa ctagcgatta gtcgctatga
      961 cttaacggag catgaaacca agctaatttt atgctgtgtg gcactactca accccacgat
     1021 tgaaaaccct acaaggaaag aacggacggt atcgttcact tataaccaat acgttcagat
     1081 gatgaacatc agtagggaaa atgcttatgg tgtattagct aaagcaacca gagagctgat
     1141 gacgagaact gtggaaatca ggaatccttt ggttaaaggc tttgagattt tccagtggac
     1201 aaactatgcc aagttctcaa gcgaaaaatt agaattagtt tttagtgaag agatattgcc
     1261 ttatcttttc cagttaaaaa aattcataaa atataatctg gaacatgtta agtcttttga
     1321 aaacaaatac tctatgagga tttatgagtg gttattaaaa gaactaacac aaaagaaaac
     1381 tcacaaggca aatatagaga ttagccttga tgaatttaag ttcatgttaa tgcttgaaaa
     1441 taactaccat gagtttaaaa ggcttaacca atgggttttg aaaccaataa gtaaagattt
     1501 aaacacttac agcaatatga aattggtggt tgataagcga ggccgcccga ctgatacgtt
     1561 gattttccaa gttgaactag atagacaaat ggatctcgta accgaacttg agaacaacca
     1621 gataaaaatg aatggtgaca aaataccaac aaccattaca tcagattcct acctacataa
     1681 cggactaaga aaaacactac acgatgcttt aactgcaaaa attcagctca ccagttttga
     1741 ggcaaaattt ttgagtgaca tgcaaagtaa gtatgatctc aatggttcgt tctcatggct
     1801 cacgcaaaaa caacgaacca cactagagaa catactggct aaatacggaa ggatctgagg
     1861 ttcttatggc tcttgtatct atcagtgaag catcaagact aacaaacaaa agtagaacaa
     1921 ctgttcaccg ttacatatca aagggaaaac tgtccatacc catgtaagaa aaaaaagaat
     1981 atataaggct tttaaagctt ttaaggttta acggttgtgg acaacaagcc agggatgtaa
     2041 cgcactgaga agcccttaga gcctctcaaa gcaattttga gtgacacagg aacacttaac
     2101 ggctgacaac cggctggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2161 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2221 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2281 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2341 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2401 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2461 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2521 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2581 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     2641 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     2701 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     2761 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     2821 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     2881 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     2941 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3001 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3061 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3121 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3181 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3241 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3301 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3361 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3421 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3481 cgtacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     3541 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     3601 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     3661 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     3721 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     3781 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
     3841 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctaga ggaccagccg
     3901 cgtaacctgg caaaatcggt tacggttgag taataaatgg atgccctgcg taagcgggtg
     3961 taggcggaca ataaagtctt aaactgaaca aaatagatct aaactatgac aataaagtct
     4021 taaactagac agaatagttg taaactgaaa tcagtccagt tatgctgtga aaaagcatac
     4081 tggacttttg ttatggctaa agcaaactct tcattttctg aagtgcaaat tgcccgtcgt
     4141 attaaagagg ggcgtggggt tcgaggtcga cggtatcgat aagctagctt aattagctga
     4201 gcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg atttgttcag
     4261 aacgctcggt tgccgccggg cgttttttat tggtgagaat ccaagctaga ctgcgatgag
     4321 tggcagggcg gggcgtaatt tttttaaggc agttattggt gcccttaaac gcctggggta
     4381 atgactctct agcttgaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     4441 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgctagg
     4501 agcttgcggc ccggacgata tcatgcatga cgatcgaatt ggggatcttg aagtacctat
     4561 tccgaagttc ctattctcta gaaagtatag gaacttcaga gcgcttttga agctgatgtg
     4621 cttaaaaact tactcaatgg aattctagat cgaattggcc gcggcgttgt gacaatttac
     4681 cgaacaactc cgcggccggg aagccgatct cggcttgaac gaattgttag gtggcggtac
     4741 ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc caactttgta tagagagcca
     4801 ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac ataagcacca agcgcgttgg
     4861 cctcatgctt gaggagattg atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg
     4921 agactgcgag atcatagata tagatctcac tacgcggctg ctcaaacttg ggcagaacgt
     4981 aagccgcgag agcgccaaca accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct
     5041 tactacggag caagttcccg aggtaatcgg agtccggctg atgttgggag taggtggcta
     5101 cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg catggatttg acttggtcag
     5161 ggccgagcct acatgtgcga atgatgccca tacttgagcc acctaacttt gttttagggc
     5221 gactgccctg ctgcgtaaca tcgttgctgc tgcgtaacat cgttgctgct ccataacatc
     5281 aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtaca
     5341 aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg
     5401 gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag tttacgaacc
     5461 gaacaggctt atgtcaattc gatccattgc tgttgacaaa gggaatcagg ggatcttgaa
     5521 gttcctattc cgaagttcct attctctaga aagtatagga acttcagagc gcttttgaag
     5581 ctaattcgga gctcactagt ggatccccaa ttcatcgatt tgacaattaa tcatcggctc
     5641 gtataatgtg tggaggatcc ctagaaataa ttttgtttaa ctttaagaag gagatataca
     5701 taatgaagcc gctgcgtggt gccgtgttta gcctgcagaa acagcatccc gactatcccg
     5761 atatctatgg cgcgattgat cagaatggga cctatcaaaa tgtgcgtacc ggcgaagatg
     5821 gtaaactgac ctttaagaat ctgagcgatg gcaaatatcg cctgtttgaa aatagcgaac
     5881 ccgctggcta taaaccggtg cagaataagc cgattgtggc gtttcagatt gtgaatggcg
     5941 aagtgcgtga tgtgaccagc attgtgccgc aggatattcc ggctacatat gaatttacca
     6001 acggtaaaca ttatatcacc aatgaaccga taccgccgaa aggatccagt ggtagcatgg
     6061 tgagcaaggg cgaggaggat aacatggcct ctctcccagc gacacatgag ttacacatct
     6121 ttggctccat caacggtgtg gactttgaca tggtgggtca gggcaccggc aatccaaatg
     6181 atggttatga ggagttaaac ctgaagtcca ccaagggtga cctccagttc tccccctgga
     6241 ttctggtccc tcatatcggg tatggcttcc atcagtacct gccctaccct gacgggatgt
     6301 cgcctttcca ggccgccatg gtagatggct ccggatacca agtccatcgc acaatgcagt
     6361 ttgaagatgg tgcctccctt actgttaact accgctacac ctacgaggga agccacatca
     6421 aaggagaggc ccaggtgaag gggactggtt tccctgctga cggtcctgtg atgaccaact
     6481 cgctgaccgc tgcggactgg tgcaggtcga agaagactta ccccaacgac aaaaccatca
     6541 tcagtacctt taagtggagt tacaccactg gaaatggcaa gcgctaccgg agcactgcgc
     6601 ggaccaccta cacctttgcc aagccaatgg cggctaacta tctgaagaac cagccgatgt
     6661 acgtgttccg taagacggag ctcaagcact ccaagaccga gctcaacttc aaggagtggc
     6721 aaaaggcctt taccgatgtg atgggcatgg acgagctgta caagtaagcg gccgcacaca
     6781 tggcatggat gagctctaca aataatgagc tagcccgcct aatgagcggg cttttttttc
     6841 tcggcctagg ggctgcagga attcctcgag aagcttgggc ccggtacctc gcgaaggcct
     6901 tgcaggccaa ccagataagt gaaatctagt tccaaactat tttgtcattt ttaattttcg
     6961 tattagctta cgacgctaca cccagttccc atctattttg tcactcttcc ctaaataatc
     7021 cttaaaaact ccatttccac ccctcccagt tcccaactat tttgtccgcc cacagcgggg
     7081 catttttctt cctgttatgt ttgggcgctg cattaatgaa tcggccaacg cgcggggaga
     7141 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgacccgct gcgctcggtc
     7201 gttcggctgc ggcgagcggt atcaggggat tccttaaggt atactttccg ctgcataacc
     7261 ctgcttcggg gtcattatag cgattttttc ggtatatcca tcctttttcg cacgatatac
     7321 aggattttgc caaagggttc gtgtagactt tccttggtgt atccaacggc gtcagccggg
     7381 caggataggt gaagtaggcc cacccgcgag cgggtgttcc ttcttcac
//
