LOCUS       40924_46793        5978 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWW3837, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5978)
  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 5978)
  AUTHORS   Whitaker WR, Shepherd ES, Sonnenburg JL.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 5978)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5978)
  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..5978
                     /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"
     terminator      complement(2373..2416)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             2569..3282
                     /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             3289..3312
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             3313..3336
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             3337..3360
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             3364..3381
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      3404..3431
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             complement(join(5389..5978,1..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 agataaaacg aaaggctcag tcgaaagact
     2401 gggcctttcg ttttacaatt gggctacctt ttttttgttt tgtttgcaat ggttaatcta
     2461 ttgttaaaat ttaaagtttc acttgaactt tcaaataatg ttcttatatt tgcagtgtcg
     2521 aaagaaacaa agtaggactg atctatggat tcaaaaaaat ttaaaataat gcgtaaaggc
     2581 gaagagctgt tcactggtgt cgtccctatt ctggtggaac tggatggtga tgtcaacggt
     2641 cataagtttt ccgtgcgtgg cgagggtgaa ggtgacgcaa ctaatggtaa actgacgctg
     2701 aagttcatct gtactactgg taaactgccg gttccttggc cgactctggt aacgacgctg
     2761 acttatggtg ttcagtgctt tgctcgttat ccggaccata tgaagcagca tgacttcttc
     2821 aagtccgcca tgccggaagg ctatgtgcag gaacgcacga tttcctttaa ggatgacggc
     2881 acgtacaaaa cgcgtgcgga agtgaaattt gaaggcgata ccctggtaaa ccgcattgag
     2941 ctgaaaggca ttgactttaa agaagacggc aatatcctgg gccataagct ggaatacaat
     3001 tttaacagcc acaatgttta catcaccgcc gataaacaaa aaaatggcat taaagcgaat
     3061 tttaaaattc gccacaacgt ggaggatggc agcgtgcagc tggctgatca ctaccagcaa
     3121 aacactccaa tcggtgatgg tcctgttctg ctgccagaca atcactatct gagcacgcaa
     3181 agcgttctgt ctaaagatcc gaacgagaaa cgcgatcata tggttctgct ggagttcgta
     3241 accgcagcgg gcatcacgca tggtatggat gaactgtaca aaggatctga ttacaaggac
     3301 gatgatgata aggactacaa agatgatgat gacaaagatt ataaggacga tgatgataaa
     3361 ggtcatcatc atcatcatca ctaaatccta gctgattagc tcgagaaggc catcctgacg
     3421 gatggccttt tttttgtatc gagacgacta gtgtaattgc ctatcttcca gtgatggaac
     3481 agcatttgtg cattggctgc aacaatcagc cttacttgtg cctgttctat ttccgaaccg
     3541 accgcttgta tgaatccatc aaaattcgtt ttctctatgt tggattcctt gttgctcata
     3601 ttgtgatgat aatttctaca aatatagtca ttggtaacta tctatgaaac tgtttgatac
     3661 ttttatagtt gattaaactt gttcatggca tttgccttaa tatcatccgc tatgtcaatg
     3721 tagggtttca tagctttgta gtcgctgtgt cccgtccatt tcatgaccac ctgtgccggg
     3781 attccgagag ccagcgcatt gcagatgaat gtccttcttc ctgcatgggt actgagcaaa
     3841 gcgtatttgg gtgtgacttc atcaatacgt tcatttccct tgtagtaggt ttcccgtaca
     3901 ggctcgttga tttctgccag ttcgcccagc tctttcaggt aatcgttcat cttctggttg
     3961 ctgatgacgg gcagagccat gtaattctcg aaatggatgt ccttgtattt gtccagtatg
     4021 gctttgctgt atttgttcag ttcaatcgtc aggctgtcgg cagtcttgac tgtggttatt
     4081 tcgatgtggt cggacttcac atcgcttctt ttcagattgc gaacatccga ataccgcaaa
     4141 ctcgtaaagc agcagaacag gaaaacatca cgcacacgtt ccaggtattg cttatccttg
     4201 ggtatctggt agtctttcag cttgttcagt tcatcccaag tcaggaagat tacttttttc
     4261 gaggtggttt tcagtttcgg tttgaacgta tcgtatgcaa tgttctgatg atgtcctttc
     4321 ttgaagctcc agcgcaggaa ccatttgagg aatcccattt gcttgccgat ggtgctgttt
     4381 ctcatatcct tggtgtcacg caggaagttg acgtattcgt tcaatccaaa ctcgttgaaa
     4441 tagttgaacg ttgcatcctc cttgaactct ttgaggtggt tcctcactgc tgcaaatttt
     4501 tcataggtgg atgccgtcca gttattctgg ttaccgcact cttttacaaa ctcatcgaac
     4561 acctcccaaa agctgacagg ggcttcttcc ggctgttctt cgctggtgtc tttcattctc
     4621 atgttgaaag cttccttcaa ctgttgggtc gttggcatga cctcctgcac ctcaaattcc
     4681 ttgaaaatat tctggatttc ggcatagtat ttcagcaagt ccgtattgat ttcggctgca
     4741 ctttgcttta gcttgttggt acatccgctc tttacccgct gcttatctgc atcccatttg
     4801 gctacgtcaa tccggtagcc cgttgtaaac tcgatgcgtt ggctggcaaa gatgacacgc
     4861 atacggatgg gtacgttctc tacgattggc acaccgttct ttttccggct ctccaatgca
     4921 aaaatgatgt tgcgcttgat attcataatt gggtgcgttt gaaattctac acccaaatat
     4981 acacccaatt attgagatag caaaagacat ttagaaacat ttacttttac tctatattgt
     5041 aatttacact tgattatcag tcgtttgcag tcttatgata ttctgtgaaa gtataagttc
     5101 gagagcctgt ctctccgcaa aaaacgctga aaatcagcag attgcaaaac aaacaccctg
     5161 ttttacaccc aagaatgtaa agtcggctgt ttttgtttta tttaagataa tacaaccact
     5221 acataataaa agagtagcga tattaaaaga atccgatgag aaaagactaa tatttatcta
     5281 tccattcagt ttgatttttc aggactttac atcgtcctga aagtatttgt tggtaccgag
     5341 gacgcgacag ctgcagacgc gttatgagta aacttggtct gacagttacc aatgcttaat
     5401 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5461 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5521 accgcgggac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5581 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5641 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5701 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5761 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5821 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5881 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5941 ctcaaccaag tcattctgag aatagtgtat gcggcgac
//
