LOCUS       40924_15250        6342 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDSG312, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6342)
  AUTHORS   Glass DS, Riedel-Kruse IH.
  TITLE     A synthetic bacterial cell-cell adhesion toolbox for programming 
            multicellular morphologies and patterns
  JOURNAL   Cell (2018) In press
REFERENCE   2  (bases 1 to 6342)
  AUTHORS   Glass DS.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-JUN-2018) Bioengineering, Stanford University, 318 
            Campus Drive, Clark Center E350, Stanford, CA 94305, USA
REFERENCE   3  (bases 1 to 6342)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6342)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell (2018)
            In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (14-JUN-2018) Bioengineering, Stanford University, 318 Campus Drive,
            Clark Center E350, Stanford, CA 94305, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6342
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4..879)
                     /label=araC
                     /note="L-arabinose regulatory protein"
     promoter        906..1190
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     misc_feature    1219..1230
                     /label=RBS (BBa_B0034)
                     /note="RBS (BBa_B0034)"
     misc_feature    1219..1230
                     /label=BBa_J04500
                     /note="BBa_J04500"
     RBS             1219..1230
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1237..3591
                     /codon_start=1
                     /product="mutated Neae"
                     /label=mutated Neae
                     /note="removed restriction sites"
                     /protein_id="AXC07666.1"
                     /translation="MITHGCYTRTRHKHKLKKTLIMLSAGLGLFFYVNQNSFANGENYF
                     KLGSDSKLLTHDSYQNRLFYTLKTGETVADLSKSQDINLSTIWSLNKHLYSSESEMMKA
                     APGQQIILPLKKLPFEYSALPLLGSAPLVAAGGVAGHTNKLTKMSPDVTKSNMTDDKAL
                     NYAAQQAASLGSQLQSRSLNGDYAKDTALGIAGNQASSQLQAWLQHYGTAEVNLQSGNN
                     FDGSSLDFLLPFYDSEKMLAFGQVGARYIDSRFTANLGAGQRFFLPANMLGYNVFIDQD
                     FSGDNTRLGIGGEYWRDYFKSSVNGYFRMSGWHESYNKKDYDERPANGFDIRFNGYLPS
                     YPALGAKLIYEQYYGDNVALFNSDKLQSNPGAATVGVNYTPIPLVTMGIDYRHGTGNEN
                     DLLYSMQFRYQFDKSWSQQIEPQYVNELRTLSGSRYDLVQRNNNIILEYKKQDILSLNI
                     PHDINGTEHSTQKIQLIVKSKYGLDRIVWDDSALRSQGGQIQHSGSQSAQDYQAILPAY
                     VQGGSNIYKVTARAYDRNGNSSNNVQLTITVLSNGQVVDQVGVTDFTADKTSAKADNAD
                     TITYTATVKKNGVAQANVPVSFNIVSGTATLGANSAKTDANGKATVTLKSSTPGQVVVS
                     AKTAEMTSALNASAVIFFDGATRQGQLVESGGGSVQAGGSLRLSCAASGIDSSSYCMGW
                     FRQRPGKEREGVARINGLGGVKTAYADSVKDRFTISRDNAENTVYLQMNSLKPEDTAIY
                     YCAAKFSPGYCGGSWSNFGYWGQGTQVTVSS"
     CDS             3211..3591
                     /codon_start=1
                     /product="EPEA tag nanobody protein"
                     /label=EPEA tag nanobody protein
                     /protein_id="AXC07668.1"
                     /translation="QGQLVESGGGSVQAGGSLRLSCAASGIDSSSYCMGWFRQRPGKER
                     EGVARINGLGGVKTAYADSVKDRFTISRDNAENTVYLQMNSLKPEDTAIYYCAAKFSPG
                     YCGGSWSNFGYWGQGTQVTVSS"
     misc_feature    3589..3594
                     /label=stop codons
                     /note="stop codons"
     misc_feature    3595..3615
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3616..3673
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E.
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     misc_feature    complement(3751..3768)
                     /label=VR primer site
                     /note="VR primer site"
     CDS             complement(3834..4646)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(5241..5785)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     misc_feature    6203..6223
                     /label=VF2 primer site
                     /note="VF2 primer site"
     terminator      complement(6275..6318)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    6321..6342
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
ORIGIN
        1 ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac ggaactcgct
       61 cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc
      121 aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg
      181 gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct ggcggaaaag
      241 atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga tatcaaaatt
      301 gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat tatccatcgg
      361 tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct caagcagatt
      421 tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga tttgcccaaa
      481 caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg tattggcaaa
      541 tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt aaacccactg
      601 gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc ctggcgggaa
      661 cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg
      721 accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt cgataaaaaa
      781 atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg cattaaacga
      841 gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac tcccgccatt
      901 cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg tcttttactg
      961 gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt aacaaagcgg
     1021 gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca
     1081 cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat ccataagatt
     1141 agcggatcct acctgacgct ttttatcgca actctctact gtttctccat acccgttttt
     1201 ttgggctagc tactagagaa agaggagaaa tactagatga ttactcatgg ttgttatacc
     1261 cggacccggc acaagcataa gctaaaaaaa acattgatta tgcttagtgc tggtttagga
     1321 ttgttttttt atgttaatca gaactcattt gcaaatggtg aaaattattt taaattgggt
     1381 tcggattcaa aactgttaac tcatgatagc tatcagaatc gcctttttta tacgttgaaa
     1441 actggtgaaa ctgttgccga tctttctaaa tcgcaagata ttaatttatc gacgatttgg
     1501 tcgttgaata agcatttata cagttctgaa agcgaaatga tgaaggccgc gcctggtcag
     1561 cagatcattt tgccactcaa aaaacttccc tttgaataca gtgcactacc acttttaggt
     1621 tcggcacctc ttgttgctgc gggtggtgtt gctggtcaca cgaataaact gactaaaatg
     1681 tccccggacg tgaccaaaag caacatgacc gatgacaagg cattaaatta tgcggcacaa
     1741 caggcggcga gtctcggtag ccagcttcag tcgcgatctc tgaacggcga ttacgcgaaa
     1801 gataccgctc ttggtatcgc tggtaaccag gcttcgtcac agttgcaggc ctggttacaa
     1861 cattatggaa cggcagaggt taatctgcaa agtggtaata actttgacgg tagttcactg
     1921 gacttcttat taccgttcta tgattccgaa aaaatgctgg catttggtca ggtcggagcg
     1981 cgttacattg actcccgctt tacggcaaat ttaggtgcgg gtcagcgttt tttccttcct
     2041 gcaaacatgt tgggctataa cgtcttcatt gatcaggatt tttctggtga taatacccgt
     2101 ttaggtattg gtggcgaata ctggcgagac tatttcaaaa gtagcgttaa cggctatttc
     2161 cgcatgagcg gctggcatga gtcatacaat aagaaagact atgatgagcg cccagcaaat
     2221 ggcttcgata tccgttttaa tggctatcta ccgtcatatc cggcattagg cgccaagctg
     2281 atatatgagc agtattatgg tgataatgtt gctttgttta attctgataa gctgcaatcg
     2341 aatcctggtg cggcgaccgt tggtgtaaac tatactccga ttcctctggt gacgatgggg
     2401 atcgattacc gtcatggtac gggtaatgaa aatgatctcc tttactcaat gcagttccgt
     2461 tatcagtttg ataaatcgtg gtctcagcaa attgaaccac agtatgttaa cgagttaaga
     2521 acattatcag gcagccgtta cgatctggtt cagcgtaata acaatattat tctggagtac
     2581 aagaagcagg atattctttc tctgaatatt ccgcatgata ttaatggtac tgaacacagt
     2641 acgcagaaga ttcagttgat cgttaagagc aaatacggtc tggatcgtat cgtctgggat
     2701 gatagtgcat tacgcagtca gggcggtcag attcagcata gcggaagcca aagcgcacaa
     2761 gactaccagg ctattttgcc tgcttatgtg caaggtggca gcaatattta taaagtgacg
     2821 gctcgcgcct atgaccgtaa tggcaatagc tctaacaatg tacagcttac tattaccgtt
     2881 ctgtcgaatg gtcaagttgt cgaccaggtt ggggtaacgg actttacggc ggataagact
     2941 tcggctaaag cggataacgc cgataccatt acttataccg cgacggtgaa aaagaatggg
     3001 gtagctcagg ctaatgtccc tgtttcattt aatattgttt caggaactgc aactcttggg
     3061 gcaaatagtg ccaaaacgga tgctaacggt aaggcaaccg taacgttgaa gtcgagtacg
     3121 ccaggacagg tcgtcgtgtc tgctaaaacc gcggagatga cttcagcact taatgccagt
     3181 gcggttatat tttttgatgg tgcgactaga caagggcagc ttgttgagag cgggggggga
     3241 agcgtacaag ccgggggatc gctgcgcttg tcgtgtgcag cttccggtat tgattccagc
     3301 tcatactgta tgggttggtt tcgtcagcgt ccgggaaagg agcgcgaggg tgtcgcacgt
     3361 atcaacgggt tgggcggcgt taaaacagct tacgctgact ctgttaaaga tcgtttcacc
     3421 attagtcgtg ataatgcaga aaatactgtt tatttgcaaa tgaactcctt gaaacccgaa
     3481 gatacagcta tttattattg tgctgccaag tttagtccag gttattgcgg aggatcgtgg
     3541 agcaatttcg gatactgggg tcaaggaacg caagtgaccg tatcttcgta ataatactag
     3601 tagcggccgc tgcagtccgg caaaaaaggg caaggtgtca ccaccctgcc ctttttcttt
     3661 aaaaccgaaa agattacttc gcgttatgca ggcttcctcg ctcactgact cgctgcgctc
     3721 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatct cgagtcccgt
     3781 caagtcagcg taatgctctg ccagtgttac aaccaattaa ccaattctga ttagaaaaac
     3841 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     3901 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     3961 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     4021 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     4081 gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc
     4141 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     4201 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     4261 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     4321 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     4381 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     4441 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     4501 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     4561 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa
     4621 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     4681 agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga
     4741 gacacaacgt ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg
     4801 catcttcccg acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg
     4861 tccacctaca acaaagctct catcaaccgt ggctccctca ctttctggct ggatgatggg
     4921 gcgattcagg cctggtatga gtcagcaaca ccttcttcac gaggcagacc tcagcgctag
     4981 cggagtgtat actggcttac tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg
     5041 tggcaggaga aaaaaggctg caccggtgcg tcagcagaat atgtgataca ggatatattc
     5101 cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc
     5161 ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag
     5221 agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc
     5281 tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5341 ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt gtcattccgc
     5401 tgttatggcc gcgtttgtct cattccacgc ctgacactca gttccgggta ggcagttcgc
     5461 tccaagctgg actgtatgca cgaacccccc gttcagtccg accgctgcgc cttatccggt
     5521 aactatcgtc ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact
     5581 ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag
     5641 gacaagtttt ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc
     5701 tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagagat
     5761 tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt aaggggtctg acgctcagtg
     5821 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5881 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5941 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6001 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6061 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6121 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6181 ctccatccag tctattccat ggtgccacct gacgtctaag aaaccattat tatcatgaca
     6241 ttaacctata aaaataggcg tatcacgagg cagaatttca gataaaaaaa atccttagct
     6301 ttcgctaagg atgatttctg gaattcgcgg ccgcttctag ag
//
