pTn7xTS-sfGFP

LOCUS       pTn7xTS-sfGFP                7181 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Tn7 tagging vector pTn7xTS with sfGFP expression scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    pTn7xTS-sfGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7181)
  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 7181)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..7181
                     /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     complement(830..850)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             complement(932..1648)
                     /codon_start=1
                     /product="enhanced blue variant of GFP"
                     /label=EBFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIKV
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(977..998)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     RBS             1656..1678
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(1702..1730)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    complement(1780..1827)
                     /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        1868..1896
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2085..2618
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    complement(2748..2795)
                     /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      2856..2942
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3045..3139
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3167..3183)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3168..3184)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3598..3617)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3717..3739
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3777..3795)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3863..3967
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3968..4828
                     /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(4186..4205)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(5487..6437)
                     /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      6485..6707
                     /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 ccctaggccg agaaaaaaaa gcccgctcat taggcgggct
      781 agctcattat ttgtagagct catccatgcc atgtgtaatc ccagcagcag ttacaaactc
      841 aagaaggacc atgtggtcac gcttttcgtt gggatctttc gaaagggcag attgtgtcga
      901 ccagctatcg atccatgggg cccgcggccg ctcacttgta cagctcgtcc atgccgagag
      961 tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt
     1021 ctttgctcag gacggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt
     1081 cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgacgt
     1141 tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga
     1201 cgttgtggct gttgaagttg tactccagct tgtgccccag gatgttgccg tcctccttga
     1261 agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg
     1321 cgcgggtctt gtaggtgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt
     1381 cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc
     1441 actgcacgcc gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg
     1501 tggtgcagat gaacttcagg gtcagcttgc cgttggtggc atcgccctcg ccctcgccgc
     1561 gcacgctgaa cttgtggccg tttacgtcgc catccagctc gaccaggatg ggcaccaccc
     1621 cggtgaacag ctcctcgccc ttgctcatat gtatatctcc ttcttaaagt taaacaaaat
     1681 tatttctagg gatcctccac acattatacg agccgatgat taattgtcaa gaattggggg
     1741 atccactagt gagctccgaa ttagcttcaa aagcgctctg aagttcctat actttctaga
     1801 gaataggaac ttcggaatag gaacttcaag atcccctgat tccctttgtc aacagcaatg
     1861 gatcgaattg acataagcct gttcggttcg taaactgtaa tgcaagtagc gtatgcgctc
     1921 acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt
     1981 catggcttgt tatgactgtt tttttgtaca gtctatgcct cgggcatcca agcagcaagc
     2041 gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcagca
     2101 acgatgttac gcagcagggc agtcgcccta aaacaaagtt aggtggctca agtatgggca
     2161 tcattcgcac atgtaggctc ggccctgacc aagtcaaatc catgcgggct gctcttgatc
     2221 ttttcggtcg tgagttcgga gacgtagcca cctactccca acatcagccg gactccgatt
     2281 acctcgggaa cttgctccgt agtaagacat tcatcgcgct tgctgccttc gaccaagaag
     2341 cggttgttgg cgctctcgcg gcttacgttc tgcccaagtt tgagcagccg cgtagtgaga
     2401 tctatatcta tgatctcgca gtctccggcg agcaccggag gcagggcatt gccaccgcgc
     2461 tcatcaatct cctcaagcat gaggccaacg cgcttggtgc ttatgtgatc tacgtgcaag
     2521 cagattacgg tgacgatccc gcagtggctc tctatacaaa gttgggcata cgggaagaag
     2581 tgatgcactt tgatatcgac ccaagtaccg ccacctaaca attcgttcaa gccgagatcg
     2641 gcttcccggc cgcggagttg ttcggtaaat tgtcacaacg ccgcggccaa ttcgatctag
     2701 aattccattg agtaagtttt taagcacatc agcttcaaaa gcgctctgaa gttcctatac
     2761 tttctagaga ataggaactt cggaataggt acttcaagat ccccaattcg atcgtcatgc
     2821 atgatatcgt ccgggccgca agctcctagc ggcggatttg tcctactcag gagagcgttc
     2881 accgacaaac aacagataaa acgaaaggcc cagtctttcg actgagcctt tcgttttatt
     2941 tgatgcctca agctagagag tcattacccc aggcgtttaa gggcaccaat aactgcctta
     3001 aaaaaattac gccccgccct gccactcatc gcagtctagc ttggattctc accaataaaa
     3061 aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact
     3121 ggatctatca acaggagtcc aagctcagct aattaagcta gcttatcgat accgtcgacc
     3181 tcgaacccca cgcccctctt taatacgacg ggcaatttgc acttcagaaa atgaagagtt
     3241 tgctttagcc ataacaaaag tccagtatgc tttttcacag cataactgga ctgatttcag
     3301 tttacaacta ttctgtctag tttaagactt tattgtcata gtttagatct attttgttca
     3361 gtttaagact ttattgtccg cctacacccg cttacgcagg gcatccattt attactcaac
     3421 cgtaaccgat tttgccaggt tacgcggctg gtcctctagc tggcgtaata gcgaagaggc
     3481 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
     3541 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
     3601 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     3661 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     3721 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     3781 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     3841 tggcactttt cggggaaatg tacgcggaac ccctatttgt ttatttttct aaatacattc
     3901 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3961 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4021 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4081 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4141 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4201 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4261 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4321 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4381 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4441 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4501 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4561 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4621 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4681 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4741 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4801 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4861 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4921 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4981 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5041 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5101 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5161 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5221 cctgttacca gccggttgtc agccgttaag tgttcctgtg tcactcaaaa ttgctttgag
     5281 aggctctaag ggcttctcag tgcgttacat ccctggcttg ttgtccacaa ccgttaaacc
     5341 ttaaaagctt taaaagcctt atatattctt ttttttctta catgggtatg gacagttttc
     5401 cctttgatat gtaacggtga acagttgttc tacttttgtt tgttagtctt gatgcttcac
     5461 tgatagatac aagagccata agaacctcag atccttccgt atttagccag tatgttctct
     5521 agtgtggttc gttgtttttg cgtgagccat gagaacgaac cattgagatc atacttactt
     5581 tgcatgtcac tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc agttaaagca
     5641 tcgtgtagtg tttttcttag tccgttatgt aggtaggaat ctgatgtaat ggttgttggt
     5701 attttgtcac cattcatttt tatctggttg ttctcaagtt cggttacgag atccatttgt
     5761 ctatctagtt caacttggaa aatcaacgta tcagtcgggc ggcctcgctt atcaaccacc
     5821 aatttcatat tgctgtaagt gtttaaatct ttacttattg gtttcaaaac ccattggtta
     5881 agccttttaa actcatggta gttattttca agcattaaca tgaacttaaa ttcatcaagg
     5941 ctaatctcta tatttgcctt gtgagttttc ttttgtgtta gttcttttaa taaccactca
     6001 taaatcctca tagagtattt gttttcaaaa gacttaacat gttccagatt atattttatg
     6061 aattttttta actggaaaag ataaggcaat atctcttcac taaaaactaa ttctaatttt
     6121 tcgcttgaga acttggcata gtttgtccac tggaaaatct caaagccttt aaccaaagga
     6181 ttcctgattt ccacagttct cgtcatcagc tctctggttg ctttagctaa tacaccataa
     6241 gcattttccc tactgatgtt catcatctga acgtattggt tataagtgaa cgataccgtc
     6301 cgttctttcc ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca gcataaaatt
     6361 agcttggttt catgctccgt taagtcatag cgactaatcg ctagttcatt tgctttgaaa
     6421 acaactaatt cagacataca tctcaattgg tctaggtgat tttaatcact ataccaattg
     6481 agatgggcta gtcaatgata attactagtc cttttccttt gagttgtggg tatctgtaaa
     6541 ttctgctaga cctttgctgg aaaacttgta aattctgcta gaccctctgt aaattccgct
     6601 agacctttgt gtgttttttt tgtttatatt caagtggtta taatttatag aataaagaaa
     6661 gaataaaaaa agataaaaag aatagatccc agccctgtgt ataactcact actttagtca
     6721 gttccgcagt attacaaaag gatgtcgcaa acgctgtttg ctcctctaca aaacagacct
     6781 taaaacccta aaggcttaag tagcaccctc gcaagctcgg ttgcggccgc aatcgggcaa
     6841 atcgctgaat attccttttg tctccgacca tcaggcacct gagtcgctgt ctttttcgtg
     6901 acattcagtt cgctgcgctc acggctctgg cagtgaatgg gggtaaatgg cactacaggc
     6961 gccttttatg gattcatgca aggaaactac ccataataca agaaaagccc gtcacgggct
     7021 tctcagggcg ttttatggcg ggtctgctat gtggtgctat ctgacttttt gctgttcagc
     7081 agttcctgcc ctctgatttt ccagtctgac cacttcggat tatcccgtga caggtcattc
     7141 agactggcta atgcacccag taaggcagcg gtatcatcaa c
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