LOCUS Exported 7338 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Tn7 tagging vector pTn7xTS with mPlum expression scaffold. ACCESSION . VERSION . KEYWORDS pTn7xTS-mPlum SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7338) AUTHORS Wiles TJ, Wall ES, Schlomann BH, Hay EA, Parthasarathy R, Guillemin K TITLE Modernized Tools for Streamlined Genetic Manipulation and Comparative Study of Wild and Diverse Proteobacterial Lineages. JOURNAL MBio. 2018 Oct 9;9(5). pii: mBio.01877-18. doi: 10.1128/mBio.01877-18. PUBMED 30301859 REFERENCE 2 (bases 1 to 7338) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7338 /organism="synthetic DNA construct" /mol_type="other DNA" oriT 106..214 /note="incP origin of transfer" mobile_element 518..683 /mobile_element_type="transposon:Tn7" /note="mini-Tn7 element (left end of the Tn7 transposon)" primer_bind 793..812 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind complement(1023..1043) /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" CDS complement(1125..1805) /codon_start=1 /product="far-red monomeric derivative of DsRed fluorescent protein (Wang et al., 2004)" /label=mPlum /note="mammalian codon-optimized" /translation="MVSKGEEVIKEFMRFKEHMEGSVNGHEFEIEGEGEGRPYEGTQTA RLKVTKGGPLPFAWDILSPQIMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGG VVTVTQDSSLQDGEFIYKVKVRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEMK MRLRLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRH STGA" primer_bind complement(1187..1210) /label=DsRed1-C /note="DsRed1, forward primer" primer_bind complement(1385..1404) /label=mCherry-F /note="mCherry, forward primer" RBS 1813..1835 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" promoter complement(1859..1887) /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind complement(1937..1984) /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 2025..2053 /gene="intI1 (promoter lies within the coding sequence)" /label=Pc promoter /note="class 1 integron promoter" CDS 2242..2775 /codon_start=1 /gene="aacC1" /product="gentamycin acetyltransferase" /label=GmR /note="confers resistance to gentamycin" /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR EEVMHFDIDPSTAT" protein_bind complement(2905..2952) /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 3013..3099 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 3202..3296 /label=lambda t0 terminator /note="transcription terminator from phage lambda" primer_bind complement(3324..3340) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3325..3341) /label=pBluescriptKS /note="For pBluescript vector" primer_bind complement(3755..3774) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3874..3896 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3934..3952) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4020..4124 /gene="bla" /label=AmpR promoter CDS 4125..4985 /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 complement(4343..4362) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" CDS complement(5644..6594) /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" rep_origin 6642..6864 /label=pSC101 ori /note="low-copy replication origin that requires the Rep101 protein" ORIGIN 1 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgtactatca 61 acaggttgaa ctgcccatgt tctttcctgc gttatcagag cttatcggcc agcctcgcag 121 agcaggattc ccgttgagca ccgccaggtg cgaataaggg acagtgaaga aggaacaccc 181 gctcgcgggt gggcctactt cacctatcct gcccggctga cgccgttgga tacaccaagg 241 aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac 301 cgaaaaaatc gctataatga ccccgaagca gggttatgca gcggaaagta taccttaagg 361 aatcccctga taccgctcgc cgcagccgaa cgaccgagcg cagcgggtca gtgagcgagg 421 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat 481 gcagcgccca aacataacag gaagaaaaat gccccgctgt gggcggacaa aatagttggg 541 aactgggagg ggtggaaatg gagtttttaa ggattattta gggaagagtg acaaaataga 601 tgggaactgg gtgtagcgtc gtaagctaat acgaaaatta aaaatgacaa aatagtttgg 661 aactagattt cacttatctg gttggcctgc aaggccttcg cgaggtaccg ggcccaagct 721 tctcgaggaa ttcctgcagc ccccggtaaa ccgaaaggca ggaacaggag agcgcacggg 781 ggagccgcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccaccactg 841 atttgagcgt cagatttcgt gatgcttgtc aggggggcgg agcctatgga aaaacggctt 901 tgccgcggcc ctctcacttc cctgttaagt atctcctagg ccgagaaaaa aaagcccgct 961 cattaggcgg gctagctcat tatttgtaga gctcatccat gccatgtgta atcccagcag 1021 cagttacaaa ctcaagaagg accatgtggt cacgcttttc gttgggatct ttcgaaaggg 1081 cagattgtgt cgaccagcta tcgatccatg gggcccgcgg ccgcctaggc gccggtggag 1141 tggcggccct cggcgcgctc gtactgttcc acgatggtgt agtcctcgtt gtgggaggtg 1201 atgtccagct tgatgtcggt cttgtaggcg ccgggcagct gcacgggctt cttggccatg 1261 taggtggtct tgacctcggc gtcgtagtgg ccgccgtcct tcagcctcag cctcatcttc 1321 atctcgccct tcagggcgcc gtcctcgggg tacatccgct cggaggaggc ctcccagccc 1381 atggtcttct tctgcattac ggggccgtcg gaggggaagt tggtgccgcg caccttcacc 1441 ttgtagatga actcgccgtc ctgcagggag gagtcctggg tcacggtcac cacgccgccg 1501 tcctcgaagt tcatcacgcg ctcccacttg aagccctcgg ggaaggacag cttcaagtag 1561 tcggggatgt cggcggggtg cttcacgtag gccttggagc cgtacatgat ctgaggggac 1621 aggatgtccc aggcgaaggg cagggggcca cccttggtca ccttcagcct ggcggtctgg 1681 gtgccctcgt aggggcggcc ctcgccctcg ccctcgatct cgaactcgtg gccgttcacg 1741 gagccctcca tgtgctcctt gaagcgcatg aactccttga tgacctcctc gcccttgctc 1801 accatatgta tatctccttc ttaaagttaa acaaaattat ttctagggat cctccacaca 1861 ttatacgagc cgatgattaa ttgtcaagaa ttgggggatc cactagtgag ctccgaatta 1921 gcttcaaaag cgctctgaag ttcctatact ttctagagaa taggaacttc ggaataggaa 1981 cttcaagatc ccctgattcc ctttgtcaac agcaatggat cgaattgaca taagcctgtt 2041 cggttcgtaa actgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga 2101 ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt 2161 ttgtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt 2221 tgatgttatg gagcagcaac gatgttacgc agcagcaacg atgttacgca gcagggcagt 2281 cgccctaaaa caaagttagg tggctcaagt atgggcatca ttcgcacatg taggctcggc 2341 cctgaccaag tcaaatccat gcgggctgct cttgatcttt tcggtcgtga gttcggagac 2401 gtagccacct actcccaaca tcagccggac tccgattacc tcgggaactt gctccgtagt 2461 aagacattca tcgcgcttgc tgccttcgac caagaagcgg ttgttggcgc tctcgcggct 2521 tacgttctgc ccaagtttga gcagccgcgt agtgagatct atatctatga tctcgcagtc 2581 tccggcgagc accggaggca gggcattgcc accgcgctca tcaatctcct caagcatgag 2641 gccaacgcgc ttggtgctta tgtgatctac gtgcaagcag attacggtga cgatcccgca 2701 gtggctctct atacaaagtt gggcatacgg gaagaagtga tgcactttga tatcgaccca 2761 agtaccgcca cctaacaatt cgttcaagcc gagatcggct tcccggccgc ggagttgttc 2821 ggtaaattgt cacaacgccg cggccaattc gatctagaat tccattgagt aagtttttaa 2881 gcacatcagc ttcaaaagcg ctctgaagtt cctatacttt ctagagaata ggaacttcgg 2941 aataggtact tcaagatccc caattcgatc gtcatgcatg atatcgtccg ggccgcaagc 3001 tcctagcggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg 3061 aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctcaagc tagagagtca 3121 ttaccccagg cgtttaaggg caccaataac tgccttaaaa aaattacgcc ccgccctgcc 3181 actcatcgca gtctagcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt 3241 tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag 3301 ctcagctaat taagctagct tatcgatacc gtcgacctcg aaccccacgc ccctctttaa 3361 tacgacgggc aatttgcact tcagaaaatg aagagtttgc tttagccata acaaaagtcc 3421 agtatgcttt ttcacagcat aactggactg atttcagttt acaactattc tgtctagttt 3481 aagactttat tgtcatagtt tagatctatt ttgttcagtt taagacttta ttgtccgcct 3541 acacccgctt acgcagggca tccatttatt actcaaccgt aaccgatttt gccaggttac 3601 gcggctggtc ctctagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac 3661 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt 3721 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt 3781 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc 3841 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt 3901 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg 3961 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtac 4021 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac 4081 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt 4141 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag 4201 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg 4261 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa 4321 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc 4381 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag 4441 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa 4501 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc 4561 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg 4621 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa 4681 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa 4741 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg 4801 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag 4861 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg 4921 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt 4981 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt 5041 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac 5101 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 5161 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 5221 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 5281 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga 5341 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagcc ggttgtcagc 5401 cgttaagtgt tcctgtgtca ctcaaaattg ctttgagagg ctctaagggc ttctcagtgc 5461 gttacatccc tggcttgttg tccacaaccg ttaaacctta aaagctttaa aagccttata 5521 tattcttttt tttcttacat gggtatggac agttttccct ttgatatgta acggtgaaca 5581 gttgttctac ttttgtttgt tagtcttgat gcttcactga tagatacaag agccataaga 5641 acctcagatc cttccgtatt tagccagtat gttctctagt gtggttcgtt gtttttgcgt 5701 gagccatgag aacgaaccat tgagatcata cttactttgc atgtcactca aaaattttgc 5761 ctcaaaactg gtgagctgaa tttttgcagt taaagcatcg tgtagtgttt ttcttagtcc 5821 gttatgtagg taggaatctg atgtaatggt tgttggtatt ttgtcaccat tcatttttat 5881 ctggttgttc tcaagttcgg ttacgagatc catttgtcta tctagttcaa cttggaaaat 5941 caacgtatca gtcgggcggc ctcgcttatc aaccaccaat ttcatattgc tgtaagtgtt 6001 taaatcttta cttattggtt tcaaaaccca ttggttaagc cttttaaact catggtagtt 6061 attttcaagc attaacatga acttaaattc atcaaggcta atctctatat ttgccttgtg 6121 agttttcttt tgtgttagtt cttttaataa ccactcataa atcctcatag agtatttgtt 6181 ttcaaaagac ttaacatgtt ccagattata ttttatgaat ttttttaact ggaaaagata 6241 aggcaatatc tcttcactaa aaactaattc taatttttcg cttgagaact tggcatagtt 6301 tgtccactgg aaaatctcaa agcctttaac caaaggattc ctgatttcca cagttctcgt 6361 catcagctct ctggttgctt tagctaatac accataagca ttttccctac tgatgttcat 6421 catctgaacg tattggttat aagtgaacga taccgtccgt tctttccttg tagggttttc 6481 aatcgtgggg ttgagtagtg ccacacagca taaaattagc ttggtttcat gctccgttaa 6541 gtcatagcga ctaatcgcta gttcatttgc tttgaaaaca actaattcag acatacatct 6601 caattggtct aggtgatttt aatcactata ccaattgaga tgggctagtc aatgataatt 6661 actagtcctt ttcctttgag ttgtgggtat ctgtaaattc tgctagacct ttgctggaaa 6721 acttgtaaat tctgctagac cctctgtaaa ttccgctaga cctttgtgtg ttttttttgt 6781 ttatattcaa gtggttataa tttatagaat aaagaaagaa taaaaaaaga taaaaagaat 6841 agatcccagc cctgtgtata actcactact ttagtcagtt ccgcagtatt acaaaaggat 6901 gtcgcaaacg ctgtttgctc ctctacaaaa cagaccttaa aaccctaaag gcttaagtag 6961 caccctcgca agctcggttg cggccgcaat cgggcaaatc gctgaatatt ccttttgtct 7021 ccgaccatca ggcacctgag tcgctgtctt tttcgtgaca ttcagttcgc tgcgctcacg 7081 gctctggcag tgaatggggg taaatggcac tacaggcgcc ttttatggat tcatgcaagg 7141 aaactaccca taatacaaga aaagcccgtc acgggcttct cagggcgttt tatggcgggt 7201 ctgctatgtg gtgctatctg actttttgct gttcagcagt tcctgccctc tgattttcca 7261 gtctgaccac ttcggattat cccgtgacag gtcattcaga ctggctaatg cacccagtaa 7321 ggcagcggta tcatcaac //