LOCUS       40924_46848        6131 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWW3867, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6131)
  AUTHORS   Whitaker WR, Shepherd ES, Sonnenburg JL.
  TITLE     Tunable expression tools enable single-cell strain distinction in 
            the gut microbiome
  JOURNAL   Cell (2017) In press
REFERENCE   2  (bases 1 to 6131)
  AUTHORS   Whitaker WR, Shepherd ES, Sonnenburg JL.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 6131)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6131)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell (2017)
            In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-MAR-2017) Microbiology "
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1560..1948)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     oriT            2119..2228
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             2716..3429
                     /codon_start=1
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             3436..3459
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             3460..3483
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             3484..3507
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             3511..3528
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      3551..3578
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             complement(join(5542..6131,1..268))
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
       61 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
      121 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
      181 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
      241 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatcatgg ccgcgggatt
      301 aaaagtcggg gattggtgaa caaaaaggtg tttctctctt taagagaaat atcgttttgc
      361 taaacagttg atattgaggt atcattttat cgtaaaagac atttttgctc aacaattgct
      421 tgacggaaat caacaaattt tagcattttg taaaaaagtc gctatataat ttggtgaatt
      481 ggagttattt tcatattttt gcatcccgaa gagtttctct taaagagaga aacatctttt
      541 gcataccttt tccgaccgaa tttttatgtc gtaaagaggg gctttgcagg gggtggactc
      601 agaaagatga gaatagatga ctattgtagt tgaaacacat agaaagttgc tgatatacag
      661 accgatacgc atatcgggat gaaccatgag tacgttcttt tctcaaaaaa cataaatatt
      721 cgaaaagaga tgcaataaat taaggagagg ttataatgaa caaagtaaat ataaaagata
      781 gtcaaaattt tattacttca aaatatcaca tagaaaaaat aatgaattgc ataagtttag
      841 atgaaaaaga taacatcttt gaaataggtg cagggaaagg tcattttact gctggattgg
      901 taaagagatg taattttgta acggcgatag aaattgattc taaattatgt gaggtaactc
      961 gtaataagct cttaaattat cctaactatc aaatagtaaa tgatgatata ctgaaattta
     1021 catttcctag ccacaatcca tataaaatat ttggcagcat accttacaac ataagcacaa
     1081 atataattcg aaaaattgtt tttgaaagtt cagccacaat aagttattta atagtggaat
     1141 atggttttgc taaaatgtta ttagatacaa acagatcact agcattgctg ttaatggcag
     1201 aggtagatat ttctatatta gcaaaaattc ctaggtatta tttccatcca aaacctaaag
     1261 tggatagcac attaattgta ttaaaaagaa agccagcaaa aatggcattt aaagagagaa
     1321 aaaaatatga aacttttgta atgaaatggg ttaacaaaga gtacgaaaaa ctgtttacaa
     1381 aaaatcaatt taataaagct ttaaaacatg cgagaatata tgatataaac aatattagtt
     1441 tcgaacaatt tgtatcgcta tttaatagtt ataaaatatt taacggctaa aaacaatagg
     1501 ccacatgcaa ctgtaaatgt ttacgcgggt accgacaccg cggtggaggg gaattcccat
     1561 gtcagccgtt aagtgttcct gtgtcactca aaattgcttt gagaggctct aagggcttct
     1621 cagtgcgtta catccctggc ttgttgtcca caaccgttaa accttaaaag ctttaaaagc
     1681 cttatatatt cttttttttc ttataaaact taaaacctta gaggctattt aagttgctga
     1741 tttatattaa ttttattgtt caaacatgag agcttagtac gtgaaacatg agagcttagt
     1801 acgttagcca tgagagctta gtacgttagc catgagggtt tagttcgtta aacatgagag
     1861 cttagtacgt taaacatgag agcttagtac gtgaaacatg agagcttagt acgtactatc
     1921 aacaggttga actgctgatc ttcagatcct ctacgccgga cgcatcgtgg ccggatcaat
     1981 tccgttttcc gctgcataac cctgcttcgg ggtcattata gcgatttttt cggtatatcc
     2041 atcctttttc gcacgatata caggattttg ccaaagggtt cgtgtagact ttccttggtg
     2101 tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc
     2161 cttcttcact gtcccttatt cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg
     2221 ctggccggct accgccggcg taacagatga gggcaagcgg atggctgatg aaaccaagcc
     2281 aaccaggaag ggcagcccac ctatcacgga attgatcccc ctcgaattgg aattcgttcc
     2341 gtctcgattc agatctaaag gatccaatct agatgatctg gaagaagcaa tgaaagctgc
     2401 tgttaagtct ccgaatcagg tattgttcct gacaggtgta ttcccatccg gtaaacgcgg
     2461 atactttgca gttgatctga ctcaggaata aattataaat taaggtaaga agattgtagg
     2521 ataagctaat gaaatagaaa aaggatgccg tcacacaact tgtcggcatt cttttttgtt
     2581 ttattagttg aaaatatagt gaaaaagttg cctaaatatg tatgttaaca aattatttgt
     2641 cgtaactttg cactccaaat ctgtttttaa catatggcac tagactgatc tatggattca
     2701 aaaaaattta aaataatgcg taaaggcgaa gagctgttca ctggtgtcgt ccctattctg
     2761 gtggaactgg atggtgatgt caacggtcat aagttttccg tgcgtggcga gggtgaaggt
     2821 gacgcaacta atggtaaact gacgctgaag ttcatctgta ctactggtaa actgccggtt
     2881 ccttggccga ctctggtaac gacgctgact tatggtgttc agtgctttgc tcgttatccg
     2941 gaccatatga agcagcatga cttcttcaag tccgccatgc cggaaggcta tgtgcaggaa
     3001 cgcacgattt cctttaagga tgacggcacg tacaaaacgc gtgcggaagt gaaatttgaa
     3061 ggcgataccc tggtaaaccg cattgagctg aaaggcattg actttaaaga agacggcaat
     3121 atcctgggcc ataagctgga atacaatttt aacagccaca atgtttacat caccgccgat
     3181 aaacaaaaaa atggcattaa agcgaatttt aaaattcgcc acaacgtgga ggatggcagc
     3241 gtgcagctgg ctgatcacta ccagcaaaac actccaatcg gtgatggtcc tgttctgctg
     3301 ccagacaatc actatctgag cacgcaaagc gttctgtcta aagatccgaa cgagaaacgc
     3361 gatcatatgg ttctgctgga gttcgtaacc gcagcgggca tcacgcatgg tatggatgaa
     3421 ctgtacaaag gatctgatta caaggacgat gatgataagg actacaaaga tgatgatgac
     3481 aaagattata aggacgatga tgataaaggt catcatcatc atcatcacta aatcctagct
     3541 gattagctcg agaaggccat cctgacggat ggcctttttt ttgtatcgag acgactagtg
     3601 taattgccta tcttccagtg atggaacagc atttgtgcat tggctgcaac aatcagcctt
     3661 acttgtgcct gttctatttc cgaaccgacc gcttgtatga atgcatcaaa attcgttttc
     3721 tctacgttgg attccttgtt gctcatattg tgatgataat ttctacaaat atagtcattg
     3781 gtaactatct atgaaactgt ttgatacttt tatagttgat taaacttgtt catggcattt
     3841 gccttaatat catccgctat gtcaatgtag ggtttcatag ctttgtagtc gctgtgtccc
     3901 gtccatttca tgaccacctg tgccgggatt ccgagagcca gcgcattgca gatgaatgtc
     3961 cttcttcctg catgggtact gagcaaagcg tatttgggtg tgacttcatc aatacgttca
     4021 tttcccttgt agtaggtttc ccgtacaggc tcgttgattt ctgccagttc gcccagctct
     4081 ttcaggtaat cgttcatctt ctggttgctg atgacgggca gagccatgta attctcgaaa
     4141 tggatgtcct tgtatttgtc cagtatggct ttgctgtatt tgttcagttc aatcgtcagg
     4201 ctgtcggcag tcttgactgt ggttatttcg atgtggtcgg acttcacatc gcttcttttc
     4261 agattgcgaa catccgaata ccgcaaactc gtaaagcagc agaacaggaa aacatcacgc
     4321 acacgttcca ggtattgctt atccttgggt atctggtagt ctttcagctt gttcagttca
     4381 tcccaagtca ggaagattac ttttttcgag gtggttttca gtttcggttt gaacgtatcg
     4441 tatgcaatgt tctgatgatg tcctttcttg aagctccagc gcaggaacca tttgaggaat
     4501 cccatttgct tgccgatggt gctgtttctc atatccttgg tgtcacgcag gaagttgacg
     4561 tattcgttca atccaaactc gttgaaatag ttgaacgttg catcctcctt gaactctttg
     4621 aggtggttcc tcactgctgc aaatttttca taggtggatg ccgtccagtt attctggtta
     4681 ccgcactctt ttacaaactc atcgaacacc tcccaaaagc tgacaggggc ttcttccggc
     4741 tgttcttcgc tggtgtcttt cattctcatg ttgaaagctt ccttcaactg ttgggtcgtt
     4801 ggcatgacct cctgcacctc aaattccttg aaaatattct ggatttcggc atagtatttc
     4861 agcaagtccg tattgatttc ggctgcactt tgctttagct tgttggtaca tccgctcttt
     4921 acccgctgct tatctgcatc ccatttggct acgtcaatcc ggtagcccgt tgtaaactcg
     4981 atgcgttggc tggcaaagat gacacgcata cggatgggta cgttctctac gattggcaca
     5041 ccgttctttt tccggctctc caatgcaaaa atgatgttgc gcttgatatt cataattggg
     5101 tgcgtttgaa attctacacc caaatataca cccaattatt gagatagcaa aagacattta
     5161 gaaacattta cttttactct atattgtaat ttacacttga ttatcagtcg tttgcagtct
     5221 tatgatattc tgtgaaagta taagttcgag agcctgtctc tccgcaaaaa acgctgaaaa
     5281 tcagcagatt gcaaaacaaa caccctgttt tacacccaag aatgtaaagt cggctgtttt
     5341 tgttttattt aagataatac aaccactaca taataaaaga gtagcgatat taaaagaatc
     5401 cgatgagaaa agactaatat ttatctatcc attcagtttg atttttcagg actttacatc
     5461 gtcctgaaag tatttgttgg taccggtacc gaggacgcga cagctgcaga cgcgttatga
     5521 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5581 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5641 gggcttacca tctggcccca gtgctgcaat gataccgcgg gacccacgct caccggctcc
     5701 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5761 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5821 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5881 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5941 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6001 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6061 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6121 tatgcggcga c
//
