LOCUS       40924_46698        6304 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWW3319, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6304)
  AUTHORS   Whitaker WR, Shepherd ES, Sonnenburg JL.
  TITLE     Tunable Expression Tools Enable Single-Cell Strain Distinction in 
            the Gut Microbiome
  JOURNAL   Cell 169 (3), 538-546 (2017)
  PUBMED    28431251
REFERENCE   2  (bases 1 to 6304)
  AUTHORS   Whitaker WR, Shepherd ES, Sonnenburg JL.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 6304)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6304)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell"; 
            date: "2017"; volume: "169"; issue: "3"; pages: "538-546"
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..6304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(29..417)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     oriT            588..697
                     /label=oriT
                     /note="incP origin of transfer"
     regulatory      841..933
                     /regulatory_class="promoter"
     terminator      1056..1127
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1143..1170
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             1364..2077
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
     CDS             2084..2107
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             2108..2131
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             2132..2155
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             2159..2176
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      2199..2226
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             complement(2458..3741)
                     /codon_start=1
                     /product="NBU integrase"
                     /label=NBU integrase
                     /protein_id="ARI71317.1"
                     /translation="MNIKRNIIFALESRKKNGVPIVENVPIRMRVIFASQRIEFTTGYR
                     IDVAKWDADKQRVKSGCTNKLKQSAAEINTDLLKYYAEIQNIFKEFEVQEVMPTTQQLK
                     EAFNMRMKDTSEEQPEEAPVSFWEVFDEFVKECGNQNNWTASTYEKFAAVRNHLKEFKE
                     DATFNYFNEFGLNEYVNFLRDTKDMRNSTIGKQMGFLKWFLRWSFKKGHHQNIAYDTFK
                     PKLKTTSKKVIFLTWDELNKLKDYQIPKDKQYLERVRDVFLFCCFTSLRYSDVRNLKRS
                     DVKSDHIEITTVKTADSLTIELNKYSKAILDKYKDIHFENYMALPVISNQKMNDYLKEL
                     GELAEINEPVRETYYKGNERIDEVTPKYALLSTHAGRRTFICNALALGIPAQVVMKWTG
                     HSDYKAMKPYIDIADDIKANAMNKFNQL"
     misc_feature    3901..3912
                     /label=integration site
                     /note="integration site"
     CDS             complement(4184..5041)
                     /label=AmpR
                     /note="beta-lactamase"
     CDS             5529..6263
                     /codon_start=1
                     /product="ErmG"
                     /label=ErmG
                     /protein_id="ARI71320.1"
                     /translation="MNKVNIKDSQNFITSKYHIEKIMNCISLDEKDNIFEIGAGKGHFT
                     AGLVKRCNFVTAIEIDSKLCEVTRNKLLNYPNYQIVNDDILKFTFPSHNPYKIFGSIPY
                     NISTNIIRKIVFESSATISYLIVEYGFAKMLLDTNRSLALLLMAEVDISILAKIPRYYF
                     HPKPKVDSTLIVLKRKPAKMAFKERKKYETFVMKWVNKEYEKLFTKNQFNKALKHARIY
                     DINNISFEQFVSLFNSYKIFNG"
ORIGIN
        1 ccgacaccgc ggtggagggg aattcccatg tcagccgtta agtgttcctg tgtcactcaa
       61 aattgctttg agaggctcta agggcttctc agtgcgttac atccctggct tgttgtccac
      121 aaccgttaaa ccttaaaagc tttaaaagcc ttatatattc ttttttttct tataaaactt
      181 aaaaccttag aggctattta agttgctgat ttatattaat tttattgttc aaacatgaga
      241 gcttagtacg tgaaacatga gagcttagta cgttagccat gagagcttag tacgttagcc
      301 atgagggttt agttcgttaa acatgagagc ttagtacgtt aaacatgaga gcttagtacg
      361 tgaaacatga gagcttagta cgtactatca acaggttgaa ctgctgatct tcagatcctc
      421 tacgccggac gcatcgtggc cggatcaatt ccgttttccg ctgcataacc ctgcttcggg
      481 gtcattatag cgattttttc ggtatatcca tcctttttcg cacgatatac aggattttgc
      541 caaagggttc gtgtagactt tccttggtgt atccaacggc gtcagccggg caggataggt
      601 gaagtaggcc cacccgcgag cgggtgttcc ttcttcactg tcccttattc gcacctggcg
      661 gtgctcaacg ggaatcctgc tctgcgaggc tggccggcta ccgccggcgt aacagatgag
      721 ggcaagcgga tggctgatga aaccaagcca accaggaagg gcagcccacc tatcacggaa
      781 ttgatccccc tcgaattgga attcgttccg tctcgattca gatctaaagg atccaatcta
      841 gagtttgcaa tggttaatct attgttaaaa tttaaagttt cacttgaact ttcaaataat
      901 gttcttatat ttgcagtgtc gaaagaaaca aaggacttat gccaacacaa ttaacatctc
      961 aatcaaggta aatgcttttt gctttttttg ctcggccggc ttgtcgacga cggccgtctc
     1021 cgtcgtcagg atcatccggg ccatatgcca ggcatcaaat aaaacgaaag gctcagtcga
     1081 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca
     1141 ctggctcacc ttcgggtggg cctttctgcg tttataatgc atgggtttgt accgtacacc
     1201 actgagacgg cggtggttga ccagacaaac cacgaaaagg agttttttag ttaccttaat
     1261 tgaaataaac gaaataaaaa ctcgcacaca acatttaaaa aataacataa tgaaagcaat
     1321 tttcgtactg aaacatctta atcatgcaca ggagattaat tcaatgcgta aaggcgaaga
     1381 gctgttcact ggtgtcgtcc ctattctggt ggaactggat ggtgatgtca acggtcataa
     1441 gttttccgtg cgtggcgagg gtgaaggtga cgcaactaat ggtaaactga cgctgaagtt
     1501 catctgtact actggtaaac tgccggttcc ttggccgact ctggtaacga cgctgactta
     1561 tggtgttcag tgctttgctc gttatccgga ccatatgaag cagcatgact tcttcaagtc
     1621 cgccatgccg gaaggctatg tgcaggaacg cacgatttcc tttaaggatg acggcacgta
     1681 caaaacgcgt gcggaagtga aatttgaagg cgataccctg gtaaaccgca ttgagctgaa
     1741 aggcattgac tttaaagaag acggcaatat cctgggccat aagctggaat acaattttaa
     1801 cagccacaat gtttacatca ccgccgataa acaaaaaaat ggcattaaag cgaattttaa
     1861 aattcgccac aacgtggagg atggcagcgt gcagctggct gatcactacc agcaaaacac
     1921 tccaatcggt gatggtcctg ttctgctgcc agacaatcac tatctgagca cgcaaagcgt
     1981 tctgtctaaa gatccgaacg agaaacgcga tcatatggtt ctgctggagt tcgtaaccgc
     2041 agcgggcatc acgcatggta tggatgaact gtacaaagga tctgattaca aggacgatga
     2101 tgataaggac tacaaagatg atgatgacaa agattataag gacgatgatg ataaaggtca
     2161 tcatcatcat catcactaaa tcctagctga ttagctcgag aaggccatcc tgacggatgg
     2221 cctttttttt gtatcgagac gactagtgta attgcctatc ttccagtgat ggaacagcat
     2281 ttgtgcattg gctgcaacaa tcagccttac ttgtgcctgt tctatttccg aaccgaccgc
     2341 ttgtatgaat ccatcaaaat tcgttttctc tatgttggat tccttgttgc tcatattgtg
     2401 atgataattt ctacaaatat agtcattggt aactatctat gaaactgttt gatactttta
     2461 tagttgatta aacttgttca tggcatttgc cttaatatca tccgctatgt caatgtaggg
     2521 tttcatagct ttgtagtcgc tgtgtcccgt ccatttcatg accacctgtg ccgggattcc
     2581 gagagccagc gcattgcaga tgaatgtcct tcttcctgca tgggtactga gcaaagcgta
     2641 tttgggtgtg acttcatcaa tacgttcatt tcccttgtag taggtttccc gtacaggctc
     2701 gttgatttct gccagttcgc ccagctcttt caggtaatcg ttcatcttct ggttgctgat
     2761 gacgggcaga gccatgtaat tctcgaaatg gatgtccttg tatttgtcca gtatggcttt
     2821 gctgtatttg ttcagttcaa tcgtcaggct gtcggcagtc ttgactgtgg ttatttcgat
     2881 gtggtcggac ttcacatcgc ttcttttcag attgcgaaca tccgaatacc gcaaactcgt
     2941 aaagcagcag aacaggaaaa catcacgcac acgttccagg tattgcttat ccttgggtat
     3001 ctggtagtct ttcagcttgt tcagttcatc ccaagtcagg aagattactt ttttcgaggt
     3061 ggttttcagt ttcggtttga acgtatcgta tgcaatgttc tgatgatgtc ctttcttgaa
     3121 gctccagcgc aggaaccatt tgaggaatcc catttgcttg ccgatggtgc tgtttctcat
     3181 atccttggtg tcacgcagga agttgacgta ttcgttcaat ccaaactcgt tgaaatagtt
     3241 gaacgttgca tcctccttga actctttgag gtggttcctc actgctgcaa atttttcata
     3301 ggtggatgcc gtccagttat tctggttacc gcactctttt acaaactcat cgaacacctc
     3361 ccaaaagctg acaggggctt cttccggctg ttcttcgctg gtgtctttca ttctcatgtt
     3421 gaaagcttcc ttcaactgtt gggtcgttgg catgacctcc tgcacctcaa attccttgaa
     3481 aatattctgg atttcggcat agtatttcag caagtccgta ttgatttcgg ctgcactttg
     3541 ctttagcttg ttggtacatc cgctctttac ccgctgctta tctgcatccc atttggctac
     3601 gtcaatccgg tagcccgttg taaactcgat gcgttggctg gcaaagatga cacgcatacg
     3661 gatgggtacg ttctctacga ttggcacacc gttctttttc cggctctcca atgcaaaaat
     3721 gatgttgcgc ttgatattca taattgggtg cgtttgaaat tctacaccca aatatacacc
     3781 caattattga gatagcaaaa gacatttaga aacatttact tttactctat attgtaattt
     3841 acacttgatt atcagtcgtt tgcagtctta tgatattctg tgaaagtata agttcgagag
     3901 cctgtctctc cgcaaaaaac gctgaaaatc agcagattgc aaaacaaaca ccctgtttta
     3961 cacccaagaa tgtaaagtcg gctgtttttg ttttatttaa gataatacaa ccactacata
     4021 ataaaagagt agcgatatta aaagaatccg atgagaaaag actaatattt atctatccat
     4081 tcagtttgat ttttcaggac tttacatcgt cctgaaagta tttgttggta ccgaggacgc
     4141 gacagctgca gacgcgttat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4201 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4261 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4321 gggacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4381 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4441 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4501 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4561 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4621 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4681 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4741 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4801 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4861 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4921 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4981 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5041 tactcttcct ttttcaatca tggccgcggg attaaaagtc ggggattggt gaacaaaaag
     5101 gtgtttctct ctttaagaga aatatcgttt tgctaaacag ttgatattga ggtatcattt
     5161 tatcgtaaaa gacatttttg ctcaacaatt gcttgacgga aatcaacaaa ttttagcatt
     5221 ttgtaaaaaa gtcgctatat aatttggtga attggagtta ttttcatatt tttgcatccc
     5281 gaagagtttc tcttaaagag agaaacatct tttgcatacc ttttccgacc gaatttttat
     5341 gtcgtaaaga ggggctttgc agggggtgga ctcagaaaga tgagaataga tgactattgt
     5401 agttgaaaca catagaaagt tgctgatata cagaccgata cgcatatcgg gatgaaccat
     5461 gagtacgttc ttttctcaaa aaacataaat attcgaaaag agatgcaata aattaaggag
     5521 aggttataat gaacaaagta aatataaaag atagtcaaaa ttttattact tcaaaatatc
     5581 acatagaaaa aataatgaat tgcataagtt tagatgaaaa agataacatc tttgaaatag
     5641 gtgcagggaa aggtcatttt actgctggat tggtaaagag atgtaatttt gtaacggcga
     5701 tagaaattga ttctaaatta tgtgaggtaa ctcgtaataa gctcttaaat tatcctaact
     5761 atcaaatagt aaatgatgat atactgaaat ttacatttcc tagccacaat ccatataaaa
     5821 tatttggcag cataccttac aacataagca caaatataat tcgaaaaatt gtttttgaaa
     5881 gttcagccac aataagttat ttaatagtgg aatatggttt tgctaaaatg ttattagata
     5941 caaacagatc actagcattg ctgttaatgg cagaggtaga tatttctata ttagcaaaaa
     6001 ttcctaggta ttatttccat ccaaaaccta aagtggatag cacattaatt gtattaaaaa
     6061 gaaagccagc aaaaatggca tttaaagaga gaaaaaaata tgaaactttt gtaatgaaat
     6121 gggttaacaa agagtacgaa aaactgttta caaaaaatca atttaataaa gctttaaaac
     6181 atgcgagaat atatgatata aacaatatta gtttcgaaca atttgtatcg ctatttaata
     6241 gttataaaat atttaacggc taaaaacaat aggccacatg caactgtaaa tgtttacgcg
     6301 ggta
//
