LOCUS       sequence-423254-        9167 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Landing Pad with T7 RNAP circuit variant T15. No seed promoter. 
            pTac-Tn5 transposase. In dap auxotrophic conjugation donor. This 
            material contains 2 plasmids. Look below for more information..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9167)
  AUTHORS   Patel JR, Oh J, Wang S, Crawford JM, Isaacs FJ
  TITLE     Cross-kingdom expression of synthetic genetic elements promotes 
            discovery of metabolites in the human microbiome.
  JOURNAL   Cell. 2022 Apr 28;185(9):1487-1505.e14. doi: 
            10.1016/j.cell.2022.03.008. Epub 2022 Apr 1.
  PUBMED    35366417
REFERENCE   2  (bases 1 to 9167)
  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"; doi: 
            "10.1016/j.cell.2022.03.008"; journalName: "Cell"; date: 
            "2022-04-28- 28"; volume: "185"; issue: "9"; pages: "1487-1505.e14"
COMMENT     Sequence Label: pLP-pTac-Tn5-chlorR
FEATURES             Location/Qualifiers
     source          1..9167
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..19
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     CDS             50..709
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(116..135)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        4410..4428
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(4456..4458,4459..5169)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    5745..5763
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     promoter        join(5787..5792,5793..5808,5809..5815)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    5819..5842
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             5929..7356
                     /codon_start=1
                     /product="transposase from the bacterial Tn5 transposon 
                     (Reznikoff, 1993)"
                     /label=Tn5 transposase
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSKAAQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     promoter        complement(7360..7378)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7361..7378)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        7496..7587
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7588..7656,7657..8448)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      8452..8491
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            8638..8746
                     /note="incP origin of transfer"
     rep_origin      8765..9153
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 ctgtctctta tacacatcta aaaggatcta ttataaggag gcactcacca tggagaaaaa
       61 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
      121 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
      181 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
      241 ccgcctgatg aatgctcatc cggaattacg tatggcaatg aaagacggtg agctggtgat
      301 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
      361 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
      421 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
      481 cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
      541 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
      601 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
      661 gcttaatgaa ttacaacagt actgcgatga gtggcaggga ggagcgtaat agggataatc
      721 agaattggtt aattggttgt aacactggtc tatcattgat aggtataaat taatacgact
      781 cactaatact gaacctatca gtgatagata ccggtgatac cagcatcgtc ttgatgccct
      841 tggcagcacc ctgctaagga ggcaacaaga tgaacacgat taacatcgct aagaacgact
      901 tctctgacat cgaactggct gctatcccgt tcaacactct ggctgaccat tacggtgagc
      961 gtttggctcg cgaacagttg gcccttgagc atgagtctta cgagatgggt gaagcacgct
     1021 tccgcaagat gtttgagcgt caacttaaag ctggtgaggt tgcggataac gctgccgcca
     1081 agcctctcat cactacccta ctccctaaga tgattgcacg catcaacgac tggtttgagg
     1141 aagtgaaagc taagcgcggc aagcgcccga cagccttcca gttcctgcaa gaaatcaagc
     1201 cggaagccgt agcgtacatc accattaaga ccactctggc ttgcctaacc agtgctgaca
     1261 atacaaccgt tcaggctgta gcaagcgcaa tcggtcgggc cattgaggac gaggctcgct
     1321 tcggtcgtat ccgtgacctt gaagctaagc acttcaagaa aaacgttgag gaacaactca
     1381 acaagcgcgt agggcacgtc tacaagaaag catttatgca agttgtcgag gctgacatgc
     1441 tctctaaggg tctactcggt ggcgaggcgt ggtcttcgtg gcataaggaa gactctattc
     1501 atgtaggagt acgctgcatc gagatgctca ttgagtcaac cggaatggtt agcttgcacc
     1561 gccaaaatgc tggcgtagta ggtcaagact ctgagactat cgaactcgca cctgaatacg
     1621 ctgaggctat cgcaacccgt gcaggtgcgc tggctggcat ctctccgatg ttccaacctt
     1681 gcgtagttcc tcctaagccg tggactggca ttactggtgg tggctattgg gctaacggtc
     1741 gtcgtcctct ggcgctggtg cgtactcaca gtaagaaagc actgatgcgc tacgaagacg
     1801 tttacatgcc tgaggtgtac aaagcgatta acattgcgca aaacaccgca tggaaaatca
     1861 acaagaaagt cctagcggtc gccaacgtaa tcaccaagtg gaagcattgt ccggtcgagg
     1921 acatccctgc gattgagcgt gaagaactcc cgatgaaacc ggaagacatc gacatgaatc
     1981 ctgaggctct caccgcgtgg aaacgtgctg ccgctgctgt gtaccgcaag gacaaggctc
     2041 gcaagtctcg ccgtatcagc cttgagttca tgcttgagca agccaataag tttgctaacc
     2101 ataaggccat ctggttccct tacaacatgg actggcgcgg tcgtgtttac gctgtgtcaa
     2161 tgttcaaccc gcaaggtaac gatatgacca aaggactgct tacgctggcg aaaggtaaac
     2221 caatcagtaa ggaaggttac tactggctga aaatccacgg tgcaaactgt gcgggtgtcg
     2281 ataaggttcc gttccctgag cgcatcaagt tcattgagga aaaccacgag aacatcatgg
     2341 cttgcgctaa gtctccactg gagaacactt ggtgggctga gcaagattct ccgttctgct
     2401 tccttgcgtt ctgctttgag tacgctgggg tacagcacca cggcctgagc tataactgct
     2461 cccttccgct ggcgtttgac gggtcttgct ctggcatcca gcacttctcc gcgatgctcc
     2521 gagatgaggt aggtggtcgc gcggttaact tgcttcctag tgaaaccgtt caggacatct
     2581 acgggattgt tgctaagaaa gtcaacgaga ttctacaagc agacgcaatc aatgggaccg
     2641 ataacgaagt agttaccgtg accgatgaga acactggtga aatctctgag aaagtcaagc
     2701 tgggcactaa ggcactggct ggtcaatggc tggcttacgg tgttactcgc agtgtgacta
     2761 agcgttcagt catgacgctg gcttacgggt ccaaagagtt cggcttccgt caacaagtgc
     2821 tggaagatac cattcagcca gctattgatt ccggcaaggg tctgatgttc actcagccga
     2881 atcaggctgc tggatacatg gctaagctga tttgggaatc tgtgagcgtg acggtggtag
     2941 ctgcggttga agcaatgaac tggcttaagt ctgctgctaa gctgctggct gctgaggtca
     3001 aagataagaa gactggagag attcttcgca agcgttgcgc tgtgcattgg gtaactcctg
     3061 atggtttccc tgtgtggcag gaatacaaga agcctattca gacgcgcttg aacctgatgt
     3121 tcctcggtca gttccgcttg cagcctacca ttaacaccaa caaagatagc gagattgatg
     3181 cacacaaaca ggagtctggt atcgctccta actttgtaca cagccaagac ggtagccacc
     3241 ttcgtaagac tgtagtgtgg gcacacgaga agtacggaat cgaatctttt gcactgattc
     3301 acgactcctt cggtaccatt ccggctgacg ctgcgaacct gttcaaagca gtgcgcgaaa
     3361 ctatggttga cacatatgag tcttgtgatg tactggctga tttctacgac cagttcgctg
     3421 accagttgca cgagtctcaa ttggacaaaa tgccagcact tccggctaaa ggtaacttga
     3481 acctccgtga catcttggag tcggacttcg cgttcgcgta aaagcttgat atcgaattcc
     3541 tgcagccccg gggatcccat ggtacgcgtg ctagtaatac gactcactaa tactgaatcc
     3601 aaaccctcgt taggggagcg tctaatttta ggagatccaa aatgtcaagg ctggataaat
     3661 caaaagtaat caatagcgcg ctggaactgc tgaacgaggt cggcatcgaa ggtctgacca
     3721 cccgcaagct ggcgcaaaaa ctgggcgtcg aacaaccgac gctgtactgg cacgtaaaaa
     3781 ataagcgtgc gctgctggac gcactggcaa ttgaaatgct ggatcgtcac cacacccact
     3841 tctgtccgct ggagggtgaa tcatggcaag atttccttcg caacaacgcg aagtcatttc
     3901 gctgcgcgct gctgagccac cgcgatggag caaaagttca tctgggcacc cgcccaacgg
     3961 agaaacaata tgaaacgctg gaaaaccagc ttgccttcct gtgccagcag ggtttcagcc
     4021 ttgagaacgc gctgtacgcg ctgagcgccg taggtcactt caccctgggc tgtgttctgg
     4081 aagaccaaga acatcaagta gcaaaagaag agcgagaaac ccctacgacc gattcgatgc
     4141 cgccgctgct gcgtcaggcg attgaactgt tcgatcacca gggcgcggaa ccggcattcc
     4201 tgtttggtct ggaacttatt atatgcggcc tagaaaaaca actgaagtgc gaaagcggta
     4261 gctaacgccg aaaaccccgc ttcggcgggg ttttgccgca taacagggta atccccaact
     4321 ggggtaacct gtgagttctc tcagttgggg aaaaaaaaac cccgcccctg acagggcggg
     4381 gttttttttt ttttcgccgc gttggctagt aatacgactc actataggga gacttaagta
     4441 taaggaggaa aaaatatgag caagggcgaa gaactgttta cgggcgtggt gccgattctg
     4501 gtggaactgg atggtgatgt caatggtcac aaattcagcg tgcgcggcga aggtgaaggc
     4561 gatgcaacca atggtaaact gacgctgaag tttatttgca ccacgggtaa actgccggtt
     4621 ccgtggccga ccctggtcac cacgctgacg tatggtgttc agtgtttcag tcgttacccg
     4681 gatcacatga aacgccacga ctttttcaag tccgcgatgc cggaaggtta tgtccaagaa
     4741 cgtaccatct catttaaaga tgacggcacc tacaaaacgc gcgccgaagt gaaattcgaa
     4801 ggtgatacgc tggttaaccg tattgaactg aaaggcatcg attttaagga agacggtaat
     4861 attctgggcc ataaactgga atataacttc aattcgcaca acgtgtacat caccgcagat
     4921 aagcagaaga acggtatcaa ggctaacttc aagatccgcc ataatgtgga agatggcagc
     4981 gttcaactgg ccgaccacta tcagcaaaac accccgattg gtgatggccc ggtcctgctg
     5041 ccggacaatc attacctgag cacgcagtct gtgctgagta aagatccgaa cgaaaagcgt
     5101 gaccacatgg tcctgctgga attcgtgacc gcggccggca tcacgcacgg tatggacgaa
     5161 ctgtataaag gctcagatat atcgggcggt atggttttta ctctggaaga ttttgttggc
     5221 gattggcgtc agaccgcggg ttataatttg gatcaagtcc tggaacaggg tggcgtaagc
     5281 tctctgttcc agaacctggg tgtgagcgtg acgccgattc agcgcatcgt tctgtccggc
     5341 gagaacggtc tgaaaattga tattcatgtg atcatcccgt acgaaggcct gagcggtgac
     5401 caaatgggtc aaatcgagaa aatctttaaa gtcgtctacc cagttgacga tcaccacttc
     5461 aaggttatct tgcattacgg tacgctggtg attgatggtg tgaccccgaa tatgattgac
     5521 tatttcggcc gtccgtatga aggcattgcc gtttttgacg gtaaaaagat caccgtcacc
     5581 ggtaccctgt ggaatggcaa taagattatt gacgagcgtc tgattaaccc ggacggcagc
     5641 ctgctgttcc gcgtgaccat caacggtgtc acgggttggc gtctgtgcga gcgcatcctg
     5701 gcataacccc aactggggta acctcagagt tctctcagtt ggggagatgt gtataagaga
     5761 cagcaaaatt ttagccgcta gagctgttga caattaatca tcggctcgta taatgtgtgg
     5821 aattgtgagc ggataacaat tcaaatttgc gcgccacatt attattcata cctttgtgga
     5881 ccgtattaca aagagaagtg ttaaatcaaa caaaaaggag gattaatcat gattaccagt
     5941 gcactgcatc gtgcggcgga ttgggcgaaa agcgtgtttt ctagtgctgc gctgggtgat
     6001 ccgcgtcgta ccgcgcgtct ggtgaatgtt gcggcgcaac tggccaaata tagcggcaaa
     6061 agcattacca ttagcagcga aggcagcaaa gccgctcagg aaggcgcgta tcgttttatt
     6121 cgtaatccga acgtgagcgc ggaagcgatt cgtaaagcgg gtgccatgca gaccgtgaaa
     6181 ctggcccagg aatttccgga actgctggca attgaagata ccacctctct gagctatcgt
     6241 catcaggtgg cggaagaact gggcaaactg ggtagcattc aggataaaag ccgtggttgg
     6301 tgggtgcata gcgtgctgct gctggaagcg accacctttc gtaccgtggg cctgctgcat
     6361 caagaatggt ggatgcgtcc ggatgatccg gcggatgcgg atgaaaaaga aagcggcaaa
     6421 tggctggccg ctgctgcaac ttcgcgtctg agaatgggca gcatgatgag caacgtgatt
     6481 gcggtgtgcg atcgtgaagc ggatattcat gcgtatctgc aagataaact ggcccataac
     6541 gaacgttttg tggtgcgtag caaacatccg cgtaaagatg tggaaagcgg cctgtatctg
     6601 tatgatcacc tgaaaaacca gccggaactg ggcggctatc agattagcat tccgcagaaa
     6661 ggcgtggtgg ataaacgtgg caaacgtaaa aaccgtccgg cgcgtaaagc gagcctgagc
     6721 ctgcgtagcg gccgtattac cctgaaacag ggcaacatta ccctgaacgc ggtgctggcc
     6781 gaagaaatta atccgccgaa aggcgaaacc ccgctgaaat ggctgctgct gaccagcgag
     6841 ccggtggaaa gtctggccca agcgctgcgt gtgattgata tttataccca tcgttggcgc
     6901 attgaagaat ttcacaaagc gtggaaaacg ggtgcgggtg cggaacgtca gcgtatggaa
     6961 gaaccggata acctggaacg tatggtgagc attctgagct ttgtggcggt gcgtctgctg
     7021 caactgcgtg aatcttttac tccrccgcaa gcactgcgtg cgcagggcct gctgaaagaa
     7081 gcggaacacg ttgaaagcca gagcgcggaa accgtgctga ccccggatga atgccaactg
     7141 ctgggctatc tggataaagg caaacgcaaa cgcaaagaaa aagcgggcag cctgcaatgg
     7201 gcgtatatgg cgattgcgcg tctgggcggc tttatggata gcaaacgtac cggcattgcg
     7261 agctggggtg cgctgtggga aggttgggaa gcgctgcaaa gcaaactgga tggctttctg
     7321 gccgcgaaag acctgatggc gcagggcatt aaaatctaat ctatagtgtc acctaaatgg
     7381 accaaaacga aaaaaggccc ccctttcggg aggcctcttt tctggaattt ggtaccgagt
     7441 aatcgattta aattagtagc ccgcctaatg agcgggcttt tttttaattc ccctatttgt
     7501 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     7561 cttcaataat attgaaaaag gaagagtatg agcattcagc attttcgtgt ggcgctgatt
     7621 cctttttttg cggcgttttg cctgccggtg tttgcgcatc cggaaaccct ggtgaaagtg
     7681 aaagatgcgg aagatcaact gggtgcgcgc gtgggctata ttgaactgga tctgaacagc
     7741 ggcaaaattc tggaatcttt tcgtccggaa gaacgttttc cgatgatgag cacctttaaa
     7801 gtgctgctgt gcggtgcggt tctgagccgt gtggatgcgg gccaggaaca actgggccgt
     7861 cgtattcatt atagccagaa cgatctggtg gaatatagcc cggtgaccga aaaacatctg
     7921 accgatggca tgaccgtgcg tgaactgtgc agcgcggcga ttaccatgag cgataacacc
     7981 gcggcgaacc tgctgctgac gaccattggc ggtccgaaag aactgaccgc gtttctgcat
     8041 aacatgggcg atcatgtgac ccgtctggat cgttgggaac cggaactgaa cgaagcgatt
     8101 ccgaacgatg aacgtgatac caccatgccg gcagcaatgg cgaccaccct gcgtaaactg
     8161 ctgacgggtg agctgctgac cctggcaagt cgccagcaac tgattgattg gatggaagcg
     8221 gataaagtgg cgggtccgct gctgcgtagc gcgctgccgg ctggctggtt tattgcggat
     8281 aaaagcggtg cgggcgaacg tggcagccgt ggcattattg cggcgctggg cccggatggt
     8341 aaaccgagcc gtattgtggt gatttatacc accggcagcc aggcgacgat ggatgaacgt
     8401 aaccgtcaga ttgcggaaat tggcgcgagc ctgattaaac attggtaaac cgatacaatt
     8461 aaaggctcct tttggagcct ttttttttgg acgacccttg tccttttccg ctgcataacc
     8521 ctgcttcggg gtcattatag cgattttttc ggtatatcca tcctttttcg cacgatatac
     8581 aggattttgc caaagggttc gtgtagactt tccttggtgt atccaacggc gtcagccggg
     8641 caggataggt gaagtaggcc cacccgcgag cgggtgttcc ttcttcactg tcccttattc
     8701 gcacctggcg gtgctcaacg ggaatcctgc tctgcgaggc tggccgtagg ccggccggcg
     8761 cgccgatctg aagatcagca gttcaacctg ttgatagtac gtactaagct ctcatgtttc
     8821 acgtactaag ctctcatgtt taacgtacta agctctcatg tttaacgaac taaaccctca
     8881 tggctaacgt actaagctct catggctaac gtactaagct ctcatgtttc acgtactaag
     8941 ctctcatgtt tgaacaataa aattaatata aatcagcaac ttaaatagcc tctaaggttt
     9001 taagttttat aagaaaaaaa agaatatata aggcttttaa agcctttaag gtttaacggt
     9061 tgtggacaac aagccaggga tgtaacgcac tgagaagccc ttagagcctc tcaaagcaat
     9121 tttgagtgac acaggaacac ttaacggctg acatggggcg cgctcaa
//
