LOCUS       40924_46738        5855 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWW3515, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5855)
  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 5855)
  AUTHORS   Whitaker WR, Shepherd ES, Sonnenburg JL.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 5855)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5855)
  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..5855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      33..125
                     /regulatory_class="promoter"
     regulatory      130..214
                     /regulatory_class="ribosome_binding_site"
     CDS             212..919
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     terminator      942..969
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             complement(1201..2484)
                     /codon_start=1
                     /product="NBU integrase"
                     /label=NBU integrase
                     /protein_id="ARI71350.1"
                     /translation="MNIKRNIIFALESRKKNGVPIVENVPIRMRVIFASQRIEFTTGYR
                     IDVAKWDADKQRVKSGCTNKLKQSAAEINTDLLKYYAEIQNIFKEFEVQEVMPTTQQLK
                     EAFNMRMKDTSEEQPEEAPVSFWEVFDEFVKECGNQNNWTASTYEKFAAVRNHLKEFKE
                     DATFNYFNEFGLNEYVNFLRDTKDMRNSTIGKQMGFLKWFLRWSFKKGHHQNIAYDTFK
                     PKLKTTSKKVIFLTWDELNKLKDYQIPKDKQYLERVRDVFLFCCFTSLRYSDVRNLKRS
                     DVKSDHIEITTVKTADSLTIELNKYSKAILDKYKDIHFENYMALPVISNQKMNDYLKEL
                     GELAEINEPVRETYYKGNERIDEVTPKYALLSTHAGRRTFICNALALGIPAQVVMKWTG
                     HSDYKAMKPYIDIADDIKANAMNKFNQL"
     misc_feature    2644..2655
                     /label=integration site
                     /note="integration site"
     CDS             complement(2927..3784)
                     /label=AmpR
                     /note="beta-lactamase"
     CDS             4272..5006
                     /codon_start=1
                     /product="ErmG"
                     /label=ErmG
                     /protein_id="ARI71352.1"
                     /translation="MNKVNIKDSQNFITSKYHIEKIMNCISLDEKDNIFEIGAGKGHFT
                     AGLVKRCNFVTAIEIDSKLCEVTRNKLLNYPNYQIVNDDILKFTFPSHNPYKIFGSIPY
                     NISTNIIRKIVFESSATISYLIVEYGFAKMLLDTNRSLALLLMAEVDISILAKIPRYYF
                     HPKPKVDSTLIVLKRKPAKMAFKERKKYETFVMKWVNKEYEKLFTKNQFNKALKHARIY
                     DINNISFEQFVSLFNSYKIFNG"
     rep_origin      complement(5076..5464)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     oriT            5635..5744
                     /label=oriT
                     /note="incP origin of transfer"
ORIGIN
        1 cgtctcgatt cagatctaaa ggatccaatc tagagtttgc aatggttaat ctattgttaa
       61 aatttaaagt ttcacttgaa ctttcaaata atgttcttat atttgcagtg tcgaaagaaa
      121 caaaggactg atcacacaac atttaaaaaa taacattatg aaagcactcg aaaagagatt
      181 caaaggtaat attaacaata atttattttc aatggtttcg aaaggggaag aagataacat
      241 ggctatcatt aaggagttta tgcgcttcaa ggtgcacatg gagggctctg tcaacggcca
      301 cgagttcgaa atcgaaggtg aaggtgaagg tcgtccgtac gaaggtactc agaccgcaaa
      361 gttgaaagtt acaaaaggag gtccgttgcc tttcgcttgg gacattctga gcccgcaatt
      421 tatgtacggg agcaaagcct atgttaaaca tccggcggat attcccgatt acctgaaact
      481 gtcattcccg gaggggttca agtgggaacg cgtcatgaac tttgaagacg gtggcgtggt
      541 tacagtgact caggattcgt cccttcaaga tggcgaattt atctacaaag tcaaactgcg
      601 cgggacaaat tttccgagtg acggtcctgt tatgcagaag aagaccatgg ggtgggaagc
      661 aagctccgaa cgcatgtacc ccgaggacgg tgcactcaaa ggagaaatca agcagcgtct
      721 gaaactgaaa gatggcgggc actacgatgc agaagtgaaa accacctata aggcgaaaaa
      781 gcctgtacaa ctgccgggtg cctataatgt gaacattaaa ctggatataa cgagtcacaa
      841 cgaagactat acaatcgtcg agcagtatga gcgtgcggaa ggacgtcatt ccactggtgg
      901 aatggatgaa ctgtataagt aaatcctagc tgattagctc gagaaggcca tcctgacgga
      961 tggccttttt tttgtatcga gacgactagt gtaattgcct atcttccagt gatggaacag
     1021 catttgtgca ttggctgcaa caatcagcct tacttgtgcc tgttctattt ccgaaccgac
     1081 cgcttgtatg aatccatcaa aattcgtttt ctctatgttg gattccttgt tgctcatatt
     1141 gtgatgataa tttctacaaa tatagtcatt ggtaactatc tatgaaactg tttgatactt
     1201 ttatagttga ttaaacttgt tcatggcatt tgccttaata tcatccgcta tgtcaatgta
     1261 gggtttcata gctttgtagt cgctgtgtcc cgtccatttc atgaccacct gtgccgggat
     1321 tccgagagcc agcgcattgc agatgaatgt ccttcttcct gcatgggtac tgagcaaagc
     1381 gtatttgggt gtgacttcat caatacgttc atttcccttg tagtaggttt cccgtacagg
     1441 ctcgttgatt tctgccagtt cgcccagctc tttcaggtaa tcgttcatct tctggttgct
     1501 gatgacgggc agagccatgt aattctcgaa atggatgtcc ttgtatttgt ccagtatggc
     1561 tttgctgtat ttgttcagtt caatcgtcag gctgtcggca gtcttgactg tggttatttc
     1621 gatgtggtcg gacttcacat cgcttctttt cagattgcga acatccgaat accgcaaact
     1681 cgtaaagcag cagaacagga aaacatcacg cacacgttcc aggtattgct tatccttggg
     1741 tatctggtag tctttcagct tgttcagttc atcccaagtc aggaagatta cttttttcga
     1801 ggtggttttc agtttcggtt tgaacgtatc gtatgcaatg ttctgatgat gtcctttctt
     1861 gaagctccag cgcaggaacc atttgaggaa tcccatttgc ttgccgatgg tgctgtttct
     1921 catatccttg gtgtcacgca ggaagttgac gtattcgttc aatccaaact cgttgaaata
     1981 gttgaacgtt gcatcctcct tgaactcttt gaggtggttc ctcactgctg caaatttttc
     2041 ataggtggat gccgtccagt tattctggtt accgcactct tttacaaact catcgaacac
     2101 ctcccaaaag ctgacagggg cttcttccgg ctgttcttcg ctggtgtctt tcattctcat
     2161 gttgaaagct tccttcaact gttgggtcgt tggcatgacc tcctgcacct caaattcctt
     2221 gaaaatattc tggatttcgg catagtattt cagcaagtcc gtattgattt cggctgcact
     2281 ttgctttagc ttgttggtac atccgctctt tacccgctgc ttatctgcat cccatttggc
     2341 tacgtcaatc cggtagcccg ttgtaaactc gatgcgttgg ctggcaaaga tgacacgcat
     2401 acggatgggt acgttctcta cgattggcac accgttcttt ttccggctct ccaatgcaaa
     2461 aatgatgttg cgcttgatat tcataattgg gtgcgtttga aattctacac ccaaatatac
     2521 acccaattat tgagatagca aaagacattt agaaacattt acttttactc tatattgtaa
     2581 tttacacttg attatcagtc gtttgcagtc ttatgatatt ctgtgaaagt ataagttcga
     2641 gagcctgtct ctccgcaaaa aacgctgaaa atcagcagat tgcaaaacaa acaccctgtt
     2701 ttacacccaa gaatgtaaag tcggctgttt ttgttttatt taagataata caaccactac
     2761 ataataaaag agtagcgata ttaaaagaat ccgatgagaa aagactaata tttatctatc
     2821 cattcagttt gatttttcag gactttacat cgtcctgaaa gtatttgttg gtaccgagga
     2881 cgcgacagct gcagacgcgt tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     2941 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3001 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3061 cgcgggaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3121 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3181 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3241 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3301 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3361 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3421 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3481 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     3541 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3601 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     3661 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     3721 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     3781 tcatactctt cctttttcaa tcatggccgc gggattaaaa gtcggggatt ggtgaacaaa
     3841 aaggtgtttc tctctttaag agaaatatcg ttttgctaaa cagttgatat tgaggtatca
     3901 ttttatcgta aaagacattt ttgctcaaca attgcttgac ggaaatcaac aaattttagc
     3961 attttgtaaa aaagtcgcta tataatttgg tgaattggag ttattttcat atttttgcat
     4021 cccgaagagt ttctcttaaa gagagaaaca tcttttgcat accttttccg accgaatttt
     4081 tatgtcgtaa agaggggctt tgcagggggt ggactcagaa agatgagaat agatgactat
     4141 tgtagttgaa acacatagaa agttgctgat atacagaccg atacgcatat cgggatgaac
     4201 catgagtacg ttcttttctc aaaaaacata aatattcgaa aagagatgca ataaattaag
     4261 gagaggttat aatgaacaaa gtaaatataa aagatagtca aaattttatt acttcaaaat
     4321 atcacataga aaaaataatg aattgcataa gtttagatga aaaagataac atctttgaaa
     4381 taggtgcagg gaaaggtcat tttactgctg gattggtaaa gagatgtaat tttgtaacgg
     4441 cgatagaaat tgattctaaa ttatgtgagg taactcgtaa taagctctta aattatccta
     4501 actatcaaat agtaaatgat gatatactga aatttacatt tcctagccac aatccatata
     4561 aaatatttgg cagcatacct tacaacataa gcacaaatat aattcgaaaa attgtttttg
     4621 aaagttcagc cacaataagt tatttaatag tggaatatgg ttttgctaaa atgttattag
     4681 atacaaacag atcactagca ttgctgttaa tggcagaggt agatatttct atattagcaa
     4741 aaattcctag gtattatttc catccaaaac ctaaagtgga tagcacatta attgtattaa
     4801 aaagaaagcc agcaaaaatg gcatttaaag agagaaaaaa atatgaaact tttgtaatga
     4861 aatgggttaa caaagagtac gaaaaactgt ttacaaaaaa tcaatttaat aaagctttaa
     4921 aacatgcgag aatatatgat ataaacaata ttagtttcga acaatttgta tcgctattta
     4981 atagttataa aatatttaac ggctaaaaac aataggccac atgcaactgt aaatgtttac
     5041 gcgggtaccg acaccgcggt ggaggggaat tcccatgtca gccgttaagt gttcctgtgt
     5101 cactcaaaat tgctttgaga ggctctaagg gcttctcagt gcgttacatc cctggcttgt
     5161 tgtccacaac cgttaaacct taaaagcttt aaaagcctta tatattcttt tttttcttat
     5221 aaaacttaaa accttagagg ctatttaagt tgctgattta tattaatttt attgttcaaa
     5281 catgagagct tagtacgtga aacatgagag cttagtacgt tagccatgag agcttagtac
     5341 gttagccatg agggtttagt tcgttaaaca tgagagctta gtacgttaaa catgagagct
     5401 tagtacgtga aacatgagag cttagtacgt actatcaaca ggttgaactg ctgatcttca
     5461 gatcctctac gccggacgca tcgtggccgg atcaattccg ttttccgctg cataaccctg
     5521 cttcggggtc attatagcga ttttttcggt atatccatcc tttttcgcac gatatacagg
     5581 attttgccaa agggttcgtg tagactttcc ttggtgtatc caacggcgtc agccgggcag
     5641 gataggtgaa gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca
     5701 cctggcggtg ctcaacggga atcctgctct gcgaggctgg ccggctaccg ccggcgtaac
     5761 agatgagggc aagcggatgg ctgatgaaac caagccaacc aggaagggca gcccacctat
     5821 cacggaattg atccccctcg aattggaatt cgttc
//
