LOCUS       40924_46783        5119 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWW3827, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5119)
  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 5119)
  AUTHORS   Whitaker WR, Shepherd ES, Sonnenburg JL.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 5119)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5119)
  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..5119
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             18..731
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
     CDS             738..761
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             762..785
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             786..809
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             813..830
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      853..880
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             complement(1112..2395)
                     /codon_start=1
                     /product="NBU integrase"
                     /label=NBU integrase
                     /protein_id="ARI71377.1"
                     /translation="MNIKRNIIFALESRKKNGVPIVENVPIRMRVIFASQRIEFTTGYR
                     IDVAKWDADKQRVKSGCTNKLKQSAAEINTDLLKYYAEIQNIFKEFEVQEVMPTTQQLK
                     EAFNMRMKDTSEEQPEEAPVSFWEVFDEFVKECGNQNNWTASTYEKFAAVRNHLKEFKE
                     DATFNYFNEFGLNEYVNFLRDTKDMRNSTIGKQMGFLKWFLRWSFKKGHHQNIAYDTFK
                     PKLKTTSKKVIFLTWDELNKLKDYQIPKDKQYLERVRDVFLFCCFTSLRYSDVRNLKRS
                     DVKSDHIEITTVKTADSLTIELNKYSKAILDKYKDIHFENYMALPVISNQKMNDYLKEL
                     GELAEINEPVRETYYKGNERIDEVTPKYALLSTHAGRRTFICNALALGIPAQVVMKWTG
                     HSDYKAMKPYIDIADDIKANAMNKFNQL"
     misc_feature    2555..2566
                     /label=integration site
                     /note="integration site"
     misc_feature    2834..3431
                     /label=split antibiotic resistance marker
                     /note="split antibiotic resistance marker"
     terminator      3447..3473
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     regulatory      3448..3477
                     /regulatory_class="terminator"
     rep_origin      complement(3579..4167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        4245..4348
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             4349..5005
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     terminator      5079..5113
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 gcggccgctg gtctccaatg cgtaaaggcg aagagctgtt cactggtgtc gtccctattc
       61 tggtggaact ggatggtgat gtcaacggtc ataagttttc cgtgcgtggc gagggtgaag
      121 gtgacgcaac taatggtaaa ctgacgctga agttcatctg tactactggt aaactgccgg
      181 ttccttggcc gactctggta acgacgctga cttatggtgt tcagtgcttt gctcgttatc
      241 cggaccatat gaagcagcat gacttcttca agtccgccat gccggaaggc tatgtgcagg
      301 aacgcacgat ttcctttaag gatgacggca cgtacaaaac gcgtgcggaa gtgaaatttg
      361 aaggcgatac cctggtaaac cgcattgagc tgaaaggcat tgactttaaa gaagacggca
      421 atatcctggg ccataagctg gaatacaatt ttaacagcca caatgtttac atcaccgccg
      481 ataaacaaaa aaatggcatt aaagcgaatt ttaaaattcg ccacaacgtg gaggatggca
      541 gcgtgcagct ggctgatcac taccagcaaa acactccaat cggtgatggt cctgttctgc
      601 tgccagacaa tcactatctg agcacgcaaa gcgttctgtc taaagatccg aacgagaaac
      661 gcgatcatat ggttctgctg gagttcgtaa ccgcagcggg catcacgcat ggtatggatg
      721 aactgtacaa aggatctgat tacaaggacg atgatgataa ggactacaaa gatgatgatg
      781 acaaagatta taaggacgat gatgataaag gtcatcatca tcatcatcac taaatcctag
      841 ctgattagct cgagaaggcc atcctgacgg atggcctttt ttttgtatcg agacgactag
      901 tgtaattgcc tatcttccag tgatggaaca gcatttgtgc attggctgca acaatcagcc
      961 ttacttgtgc ctgttctatt tccgaaccga ccgcttgtat gaatccatca aaattcgttt
     1021 tctctatgtt ggattccttg ttgctcatat tgtgatgata atttctacaa atatagtcat
     1081 tggtaactat ctatgaaact gtttgatact tttatagttg attaaacttg ttcatggcat
     1141 ttgccttaat atcatccgct atgtcaatgt agggtttcat agctttgtag tcgctgtgtc
     1201 ccgtccattt catgaccacc tgtgccggga ttccgagagc cagcgcattg cagatgaatg
     1261 tccttcttcc tgcatgggta ctgagcaaag cgtatttggg tgtgacttca tcaatacgtt
     1321 catttccctt gtagtaggtt tcccgtacag gctcgttgat ttctgccagt tcgcccagct
     1381 ctttcaggta atcgttcatc ttctggttgc tgatgacggg cagagccatg taattctcga
     1441 aatggatgtc cttgtatttg tccagtatgg ctttgctgta tttgttcagt tcaatcgtca
     1501 ggctgtcggc agtcttgact gtggttattt cgatgtggtc ggacttcaca tcgcttcttt
     1561 tcagattgcg aacatccgaa taccgcaaac tcgtaaagca gcagaacagg aaaacatcac
     1621 gcacacgttc caggtattgc ttatccttgg gtatctggta gtctttcagc ttgttcagtt
     1681 catcccaagt caggaagatt acttttttcg aggtggtttt cagtttcggt ttgaacgtat
     1741 cgtatgcaat gttctgatga tgtcctttct tgaagctcca gcgcaggaac catttgagga
     1801 atcccatttg cttgccgatg gtgctgtttc tcatatcctt ggtgtcacgc aggaagttga
     1861 cgtattcgtt caatccaaac tcgttgaaat agttgaacgt tgcatcctcc ttgaactctt
     1921 tgaggtggtt cctcactgct gcaaattttt cataggtgga tgccgtccag ttattctggt
     1981 taccgcactc ttttacaaac tcatcgaaca cctcccaaaa gctgacaggg gcttcttccg
     2041 gctgttcttc gctggtgtct ttcattctca tgttgaaagc ttccttcaac tgttgggtcg
     2101 ttggcatgac ctcctgcacc tcaaattcct tgaaaatatt ctggatttcg gcatagtatt
     2161 tcagcaagtc cgtattgatt tcggctgcac tttgctttag cttgttggta catccgctct
     2221 ttacccgctg cttatctgca tcccatttgg ctacgtcaat ccggtagccc gttgtaaact
     2281 cgatgcgttg gctggcaaag atgacacgca tacggatggg tacgttctct acgattggca
     2341 caccgttctt tttccggctc tccaatgcaa aaatgatgtt gcgcttgata ttcataattg
     2401 ggtgcgtttg aaattctaca cccaaatata cacccaatta ttgagatagc aaaagacatt
     2461 tagaaacatt tacttttact ctatattgta atttacactt gattatcagt cgtttgcagt
     2521 cttatgatat tctgtgaaag tataagttcg agagcctgtc tctccgcaaa aaacgctgaa
     2581 aatcagcaga ttgcaaaaca aacaccctgt tttacaccca agaatgtaaa gtcggctgtt
     2641 tttgttttat ttaagataat acaaccacta cataataaaa gagtagcgat attaaaagaa
     2701 tccgatgaga aaagactaat atttatctat ccattcagtt tgatttttca ggactttaca
     2761 tcgtcctgaa agtatttgtt ggtaccgagg acgcgacagc tgcagacgcg ttatgagtaa
     2821 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2881 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     2941 ttaccatctg gccccagtgc tgcaatgata ccgcgggacc cacgctcacc ggctccagat
     3001 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3061 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3121 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3181 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3241 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3301 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3361 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3421 cggcgaccga gtgagaccac ctgcaggaag gccatcctga cggatggcct tttttttaga
     3481 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3541 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3601 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3661 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3721 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3781 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3841 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3901 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3961 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4021 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4081 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4141 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4201 gctcagtgga acgaaaactc acgttaaggg attttggtca tgattgatcg ggcacgtaag
     4261 aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt ttgagttatc
     4321 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
     4381 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
     4441 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
     4501 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
     4561 ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
     4621 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
     4681 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
     4741 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtttcagcca atccctgggt
     4801 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
     4861 caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
     4921 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
     4981 ctgcgatgag tggcagggcg gggcgtaatt tgatatcgag ctcgcttgga ctcctgttga
     5041 tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc
     5101 gggcgttttt tattggtag
//
