LOCUS       40924_46843        6335 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWW3866, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6335)
  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 6335)
  AUTHORS   Whitaker WR, Shepherd ES, Sonnenburg JL.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 6335)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6335)
  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..6335
                     /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             2920..3633
                     /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             3640..3663
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             3664..3687
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             3688..3711
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             3715..3732
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      3755..3782
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             complement(join(5746..6335,1..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 aggctacttt tgcacccgct ttccaagaga
     2401 agaaagcctt gataaattga cttagtgtaa aagcaagtgt ctgcttaacc ataagaacaa
     2461 aaaaacttcc gataaagttt ggaagataaa gctaaaagtt cttatctttg cagtccgatt
     2521 cgcaaagaaa aggtgttacg cttttcttct ttaccttctt tccctttcgc taagagagcc
     2581 tgaaaaacga tagaaaaaga aaaacgaaaa aaaaacttcc gaaaatattt ggtagttaaa
     2641 ataaaacctc ttacctttgc acccgctttt aaaacgaaag caagatgttc tttgaaatat
     2701 tgataaacaa tacaagtagt acaagaaaaa aatagaaccg tcaatacttg tcttatatgt
     2761 agtaatatgt atgagtcata aggtattaat gaagtcaata aattgtacgg catcctgaac
     2821 agagcaaaaa tcagctttat gctgactaac aatactttta caatgaagag tttgatcctg
     2881 gctcaggact gatctatgga ttcaaaaaaa tttaaaataa tgcgtaaagg cgaagagctg
     2941 ttcactggtg tcgtccctat tctggtggaa ctggatggtg atgtcaacgg tcataagttt
     3001 tccgtgcgtg gcgagggtga aggtgacgca actaatggta aactgacgct gaagttcatc
     3061 tgtactactg gtaaactgcc ggttccttgg ccgactctgg taacgacgct gacttatggt
     3121 gttcagtgct ttgctcgtta tccggaccat atgaagcagc atgacttctt caagtccgcc
     3181 atgccggaag gctatgtgca ggaacgcacg atttccttta aggatgacgg cacgtacaaa
     3241 acgcgtgcgg aagtgaaatt tgaaggcgat accctggtaa accgcattga gctgaaaggc
     3301 attgacttta aagaagacgg caatatcctg ggccataagc tggaatacaa ttttaacagc
     3361 cacaatgttt acatcaccgc cgataaacaa aaaaatggca ttaaagcgaa ttttaaaatt
     3421 cgccacaacg tggaggatgg cagcgtgcag ctggctgatc actaccagca aaacactcca
     3481 atcggtgatg gtcctgttct gctgccagac aatcactatc tgagcacgca aagcgttctg
     3541 tctaaagatc cgaacgagaa acgcgatcat atggttctgc tggagttcgt aaccgcagcg
     3601 ggcatcacgc atggtatgga tgaactgtac aaaggatctg attacaagga cgatgatgat
     3661 aaggactaca aagatgatga tgacaaagat tataaggacg atgatgataa aggtcatcat
     3721 catcatcatc actaaatcct agctgattag ctcgagaagg ccatcctgac ggatggcctt
     3781 ttttttgtat cgagacgact agtgtaattg cctatcttcc agtgatggaa cagcatttgt
     3841 gcattggctg caacaatcag ccttacttgt gcctgttcta tttccgaacc gaccgcttgt
     3901 atgaatgcat caaaattcgt tttctctacg ttggattcct tgttgctcat attgtgatga
     3961 taatttctac aaatatagtc attggtaact atctatgaaa ctgtttgata cttttatagt
     4021 tgattaaact tgttcatggc atttgcctta atatcatccg ctatgtcaat gtagggtttc
     4081 atagctttgt agtcgctgtg tcccgtccat ttcatgacca cctgtgccgg gattccgaga
     4141 gccagcgcat tgcagatgaa tgtccttctt cctgcatggg tactgagcaa agcgtatttg
     4201 ggtgtgactt catcaatacg ttcatttccc ttgtagtagg tttcccgtac aggctcgttg
     4261 atttctgcca gttcgcccag ctctttcagg taatcgttca tcttctggtt gctgatgacg
     4321 ggcagagcca tgtaattctc gaaatggatg tccttgtatt tgtccagtat ggctttgctg
     4381 tatttgttca gttcaatcgt caggctgtcg gcagtcttga ctgtggttat ttcgatgtgg
     4441 tcggacttca catcgcttct tttcagattg cgaacatccg aataccgcaa actcgtaaag
     4501 cagcagaaca ggaaaacatc acgcacacgt tccaggtatt gcttatcctt gggtatctgg
     4561 tagtctttca gcttgttcag ttcatcccaa gtcaggaaga ttactttttt cgaggtggtt
     4621 ttcagtttcg gtttgaacgt atcgtatgca atgttctgat gatgtccttt cttgaagctc
     4681 cagcgcagga accatttgag gaatcccatt tgcttgccga tggtgctgtt tctcatatcc
     4741 ttggtgtcac gcaggaagtt gacgtattcg ttcaatccaa actcgttgaa atagttgaac
     4801 gttgcatcct ccttgaactc tttgaggtgg ttcctcactg ctgcaaattt ttcataggtg
     4861 gatgccgtcc agttattctg gttaccgcac tcttttacaa actcatcgaa cacctcccaa
     4921 aagctgacag gggcttcttc cggctgttct tcgctggtgt ctttcattct catgttgaaa
     4981 gcttccttca actgttgggt cgttggcatg acctcctgca cctcaaattc cttgaaaata
     5041 ttctggattt cggcatagta tttcagcaag tccgtattga tttcggctgc actttgcttt
     5101 agcttgttgg tacatccgct ctttacccgc tgcttatctg catcccattt ggctacgtca
     5161 atccggtagc ccgttgtaaa ctcgatgcgt tggctggcaa agatgacacg catacggatg
     5221 ggtacgttct ctacgattgg cacaccgttc tttttccggc tctccaatgc aaaaatgatg
     5281 ttgcgcttga tattcataat tgggtgcgtt tgaaattcta cacccaaata tacacccaat
     5341 tattgagata gcaaaagaca tttagaaaca tttactttta ctctatattg taatttacac
     5401 ttgattatca gtcgtttgca gtcttatgat attctgtgaa agtataagtt cgagagcctg
     5461 tctctccgca aaaaacgctg aaaatcagca gattgcaaaa caaacaccct gttttacacc
     5521 caagaatgta aagtcggctg tttttgtttt atttaagata atacaaccac tacataataa
     5581 aagagtagcg atattaaaag aatccgatga gaaaagacta atatttatct atccattcag
     5641 tttgattttt caggacttta catcgtcctg aaagtatttg ttggtaccgg taccgaggac
     5701 gcgacagctg cagacgcgtt atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5761 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5821 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5881 gcgggaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5941 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6001 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6061 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6121 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6181 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6241 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6301 aaccaagtca ttctgagaat agtgtatgcg gcgac
//
