LOCUS       40924_46858        6320 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWW3872, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6320)
  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 6320)
  AUTHORS   Whitaker WR, Shepherd ES, Sonnenburg JL.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 6320)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6320)
  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..6320
                     /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             2905..3618
                     /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             3625..3648
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             3649..3672
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             3673..3696
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             3700..3717
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      3740..3767
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             complement(join(5731..6320,1..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 actttccctg gggtggctag cacaatgatg
     2401 gcttgccctg atatggaagt ggaaggtaag attctgaaag caatgaatga agtgaagtcg
     2461 ttcgataaac tttccggtgg tggaatcggt ttgtatgacg caaataatgc gttggtgatt
     2521 gtacttgaaa agtaagatat acgtattata ttaaataggt atgcctgtgt cgcacataac
     2581 gatacaggca tattttttat tcctttatga tctattcttt tgctttattg aaccagagct
     2641 gtttccattt gtttaatgag tcgatagctg tctggcttct atgtgacttg acagcagtca
     2701 tttcgattta aaactgtcgt tttggcagaa aatatgtcac gtattcgtcc gatttctttt
     2761 tggcacatgg tttgtttttt aatcagcgtc cgtttgaaag cccttacaaa gagaaagaaa
     2821 acaaccggac aaaaagaaat tgaataataa taaataaaaa gaataacgat cgactgatct
     2881 atggattcaa aaaaatttaa aataatgcgt aaaggcgaag agctgttcac tggtgtcgtc
     2941 cctattctgg tggaactgga tggtgatgtc aacggtcata agttttccgt gcgtggcgag
     3001 ggtgaaggtg acgcaactaa tggtaaactg acgctgaagt tcatctgtac tactggtaaa
     3061 ctgccggttc cttggccgac tctggtaacg acgctgactt atggtgttca gtgctttgct
     3121 cgttatccgg accatatgaa gcagcatgac ttcttcaagt ccgccatgcc ggaaggctat
     3181 gtgcaggaac gcacgatttc ctttaaggat gacggcacgt acaaaacgcg tgcggaagtg
     3241 aaatttgaag gcgataccct ggtaaaccgc attgagctga aaggcattga ctttaaagaa
     3301 gacggcaata tcctgggcca taagctggaa tacaatttta acagccacaa tgtttacatc
     3361 accgccgata aacaaaaaaa tggcattaaa gcgaatttta aaattcgcca caacgtggag
     3421 gatggcagcg tgcagctggc tgatcactac cagcaaaaca ctccaatcgg tgatggtcct
     3481 gttctgctgc cagacaatca ctatctgagc acgcaaagcg ttctgtctaa agatccgaac
     3541 gagaaacgcg atcatatggt tctgctggag ttcgtaaccg cagcgggcat cacgcatggt
     3601 atggatgaac tgtacaaagg atctgattac aaggacgatg atgataagga ctacaaagat
     3661 gatgatgaca aagattataa ggacgatgat gataaaggtc atcatcatca tcatcactaa
     3721 atcctagctg attagctcga gaaggccatc ctgacggatg gccttttttt tgtatcgaga
     3781 cgactagtgt aattgcctat cttccagtga tggaacagca tttgtgcatt ggctgcaaca
     3841 atcagcctta cttgtgcctg ttctatttcc gaaccgaccg cttgtatgaa tgcatcaaaa
     3901 ttcgttttct ctacgttgga ttccttgttg ctcatattgt gatgataatt tctacaaata
     3961 tagtcattgg taactatcta tgaaactgtt tgatactttt atagttgatt aaacttgttc
     4021 atggcatttg ccttaatatc atccgctatg tcaatgtagg gtttcatagc tttgtagtcg
     4081 ctgtgtcccg tccatttcat gaccacctgt gccgggattc cgagagccag cgcattgcag
     4141 atgaatgtcc ttcttcctgc atgggtactg agcaaagcgt atttgggtgt gacttcatca
     4201 atacgttcat ttcccttgta gtaggtttcc cgtacaggct cgttgatttc tgccagttcg
     4261 cccagctctt tcaggtaatc gttcatcttc tggttgctga tgacgggcag agccatgtaa
     4321 ttctcgaaat ggatgtcctt gtatttgtcc agtatggctt tgctgtattt gttcagttca
     4381 atcgtcaggc tgtcggcagt cttgactgtg gttatttcga tgtggtcgga cttcacatcg
     4441 cttcttttca gattgcgaac atccgaatac cgcaaactcg taaagcagca gaacaggaaa
     4501 acatcacgca cacgttccag gtattgctta tccttgggta tctggtagtc tttcagcttg
     4561 ttcagttcat cccaagtcag gaagattact tttttcgagg tggttttcag tttcggtttg
     4621 aacgtatcgt atgcaatgtt ctgatgatgt cctttcttga agctccagcg caggaaccat
     4681 ttgaggaatc ccatttgctt gccgatggtg ctgtttctca tatccttggt gtcacgcagg
     4741 aagttgacgt attcgttcaa tccaaactcg ttgaaatagt tgaacgttgc atcctccttg
     4801 aactctttga ggtggttcct cactgctgca aatttttcat aggtggatgc cgtccagtta
     4861 ttctggttac cgcactcttt tacaaactca tcgaacacct cccaaaagct gacaggggct
     4921 tcttccggct gttcttcgct ggtgtctttc attctcatgt tgaaagcttc cttcaactgt
     4981 tgggtcgttg gcatgacctc ctgcacctca aattccttga aaatattctg gatttcggca
     5041 tagtatttca gcaagtccgt attgatttcg gctgcacttt gctttagctt gttggtacat
     5101 ccgctcttta cccgctgctt atctgcatcc catttggcta cgtcaatccg gtagcccgtt
     5161 gtaaactcga tgcgttggct ggcaaagatg acacgcatac ggatgggtac gttctctacg
     5221 attggcacac cgttcttttt ccggctctcc aatgcaaaaa tgatgttgcg cttgatattc
     5281 ataattgggt gcgtttgaaa ttctacaccc aaatatacac ccaattattg agatagcaaa
     5341 agacatttag aaacatttac ttttactcta tattgtaatt tacacttgat tatcagtcgt
     5401 ttgcagtctt atgatattct gtgaaagtat aagttcgaga gcctgtctct ccgcaaaaaa
     5461 cgctgaaaat cagcagattg caaaacaaac accctgtttt acacccaaga atgtaaagtc
     5521 ggctgttttt gttttattta agataataca accactacat aataaaagag tagcgatatt
     5581 aaaagaatcc gatgagaaaa gactaatatt tatctatcca ttcagtttga tttttcagga
     5641 ctttacatcg tcctgaaagt atttgttggt accggtaccg aggacgcgac agctgcagac
     5701 gcgttatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5761 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5821 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcggg acccacgctc
     5881 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5941 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6001 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6061 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6121 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6181 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6241 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6301 agaatagtgt atgcggcgac
//
