LOCUS       40924_15230        6748 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDSG289, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6748)
  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 6748)
  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 6748)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6748)
  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..6748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..36
                     /label=pLacIQ (BBa_I4032, 1nt deletion)
                     /note="pLacIQ (BBa_I4032, 1nt deletion)"
     misc_feature    45..56
                     /label=RBS (BBa_B0034)(1)
                     /note="RBS (BBa_B0034)(1)"
     misc_feature    45..56
                     /label=BBa_J04500(1)
                     /note="BBa_J04500(1)"
     RBS             45..56
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             63..683
                     /label=TetR
                     /note="tetracycline repressor TetR"
     terminator      706..777
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      793..820
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    835..853
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    860..878
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    897..908
                     /label=RBS (BBa_B0034)
                     /note="RBS (BBa_B0034)"
     misc_feature    897..908
                     /label=BBa_J04500
                     /note="BBa_J04500"
     RBS             897..908
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             915..3269
                     /codon_start=1
                     /product="mutated Neae"
                     /label=mutated Neae
                     /note="removed restriction sites"
                     /protein_id="AXC07651.1"
                     /translation="MITHGCYTRTRHKHKLKKTLIMLSAGLGLFFYVNQNSFANGENYF
                     KLGSDSKLLTHDSYQNRLFYTLKTGETVADLSKSQDINLSTIWSLNKHLYSSESEMMKA
                     APGQQIILPLKKLPFEYSALPLLGSAPLVAAGGVAGHTNKLTKMSPDVTKSNMTDDKAL
                     NYAAQQAASLGSQLQSRSLNGDYAKDTALGIAGNQASSQLQAWLQHYGTAEVNLQSGNN
                     FDGSSLDFLLPFYDSEKMLAFGQVGARYIDSRFTANLGAGQRFFLPANMLGYNVFIDQD
                     FSGDNTRLGIGGEYWRDYFKSSVNGYFRMSGWHESYNKKDYDERPANGFDIRFNGYLPS
                     YPALGAKLIYEQYYGDNVALFNSDKLQSNPGAATVGVNYTPIPLVTMGIDYRHGTGNEN
                     DLLYSMQFRYQFDKSWSQQIEPQYVNELRTLSGSRYDLVQRNNNIILEYKKQDILSLNI
                     PHDINGTEHSTQKIQLIVKSKYGLDRIVWDDSALRSQGGQIQHSGSQSAQDYQAILPAY
                     VQGGSNIYKVTARAYDRNGNSSNNVQLTITVLSNGQVVDQVGVTDFTADKTSAKADNAD
                     TITYTATVKKNGVAQANVPVSFNIVSGTATLGANSAKTDANGKATVTLKSSTPGQVVVS
                     AKTAEMTSALNASAVIFFDGATRQGQLVESGGGSVQAGGSLRLSCAASGIDSSSYCMGW
                     FRQRPGKEREGVARINGLGGVKTAYADSVKDRFTISRDNAENTVYLQMNSLKPEDTAIY
                     YCAAKFSPGYCGGSWSNFGYWGQGTQVTVSS"
     CDS             2889..3269
                     /codon_start=1
                     /product="EPEA tag nanobody protein"
                     /label=EPEA tag nanobody protein
                     /protein_id="AXC07652.1"
                     /translation="QGQLVESGGGSVQAGGSLRLSCAASGIDSSSYCMGWFRQRPGKER
                     EGVARINGLGGVKTAYADSVKDRFTISRDNAENTVYLQMNSLKPEDTAIYYCAAKFSPG
                     YCGGSWSNFGYWGQGTQVTVSS"
     misc_feature    3267..3272
                     /label=stop codons
                     /note="stop codons"
     terminator      complement(3292..3335)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3376..3403
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3410..3430
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3431..3490
                     /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(3568..3585)
                     /label=VR primer site
                     /note="VR primer site"
     CDS             complement(3796..4743)
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
     misc_feature    complement(4747..4759)
                     /label=RBS for repA
                     /note="RBS for repA"
     regulatory      complement(4764..4783)
                     /label=repA promoter
                     /note="repA promoter"
                     /regulatory_class="promoter"
     rep_origin      complement(4791..5013)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5505..5599)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5632..6489)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6490..6594)
                     /label=AmpR promoter
     misc_feature    6609..6629
                     /label=VF2 primer site
                     /note="VF2 primer site"
     terminator      complement(6681..6724)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    6727..6748
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
ORIGIN
        1 tggtgcaaaa cctttgcggt atggcatgat agcgcctact agagaaagag gagaaatact
       61 agatgtccag attagataaa agtaaagtga ttaacagcgc attagagctg cttaatgagg
      121 tcggaatcga aggtttaaca acccgtaaac tcgcccagaa gctaggtgta gagcagccta
      181 cattgtattg gcatgtaaaa aataagcggg ctttgctcga cgccttagcc attgagatgt
      241 tagataggca ccatactcac ttttgccctt tagaagggga aagctggcaa gattttttac
      301 gtaataacgc taaaagtttt agatgtgctt tactaagtca tcgcgatgga gcaaaagtac
      361 atttaggtac acggcctaca gaaaaacagt atgaaactct cgaaaatcaa ttagcctttt
      421 tatgccaaca aggtttttca ctagagaatg cattatatgc actcagcgct gtggggcatt
      481 ttactttagg ttgcgtattg gaagatcaag agcatcaagt cgctaaagaa gaaagggaaa
      541 cacctactac tgatagtatg ccgccattat tacgacaagc tatcgaatta tttgatcacc
      601 aaggtgcaga gccagccttc ttattcggcc ttgaattgat catatgcgga ttagaaaaac
      661 aacttaaatg tgaaagtggg tcctaataat actagagcca ggcatcaaat aaaacgaaag
      721 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac
      781 tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttatatact agagtcccta
      841 tcagtgatag agattgacat ccctatcagt gatagagata ctgagcacta ctagagaaag
      901 aggagaaata ctagatgatt actcatggtt gttatacccg gacccggcac aagcataagc
      961 taaaaaaaac attgattatg cttagtgctg gtttaggatt gtttttttat gttaatcaga
     1021 actcatttgc aaatggtgaa aattatttta aattgggttc ggattcaaaa ctgttaactc
     1081 atgatagcta tcagaatcgc cttttttata cgttgaaaac tggtgaaact gttgccgatc
     1141 tttctaaatc gcaagatatt aatttatcga cgatttggtc gttgaataag catttataca
     1201 gttctgaaag cgaaatgatg aaggccgcgc ctggtcagca gatcattttg ccactcaaaa
     1261 aacttccctt tgaatacagt gcactaccac ttttaggttc ggcacctctt gttgctgcgg
     1321 gtggtgttgc tggtcacacg aataaactga ctaaaatgtc cccggacgtg accaaaagca
     1381 acatgaccga tgacaaggca ttaaattatg cggcacaaca ggcggcgagt ctcggtagcc
     1441 agcttcagtc gcgatctctg aacggcgatt acgcgaaaga taccgctctt ggtatcgctg
     1501 gtaaccaggc ttcgtcacag ttgcaggcct ggttacaaca ttatggaacg gcagaggtta
     1561 atctgcaaag tggtaataac tttgacggta gttcactgga cttcttatta ccgttctatg
     1621 attccgaaaa aatgctggca tttggtcagg tcggagcgcg ttacattgac tcccgcttta
     1681 cggcaaattt aggtgcgggt cagcgttttt tccttcctgc aaacatgttg ggctataacg
     1741 tcttcattga tcaggatttt tctggtgata atacccgttt aggtattggt ggcgaatact
     1801 ggcgagacta tttcaaaagt agcgttaacg gctatttccg catgagcggc tggcatgagt
     1861 catacaataa gaaagactat gatgagcgcc cagcaaatgg cttcgatatc cgttttaatg
     1921 gctatctacc gtcatatccg gcattaggcg ccaagctgat atatgagcag tattatggtg
     1981 ataatgttgc tttgtttaat tctgataagc tgcaatcgaa tcctggtgcg gcgaccgttg
     2041 gtgtaaacta tactccgatt cctctggtga cgatggggat cgattaccgt catggtacgg
     2101 gtaatgaaaa tgatctcctt tactcaatgc agttccgtta tcagtttgat aaatcgtggt
     2161 ctcagcaaat tgaaccacag tatgttaacg agttaagaac attatcaggc agccgttacg
     2221 atctggttca gcgtaataac aatattattc tggagtacaa gaagcaggat attctttctc
     2281 tgaatattcc gcatgatatt aatggtactg aacacagtac gcagaagatt cagttgatcg
     2341 ttaagagcaa atacggtctg gatcgtatcg tctgggatga tagtgcatta cgcagtcagg
     2401 gcggtcagat tcagcatagc ggaagccaaa gcgcacaaga ctaccaggct attttgcctg
     2461 cttatgtgca aggtggcagc aatatttata aagtgacggc tcgcgcctat gaccgtaatg
     2521 gcaatagctc taacaatgta cagcttacta ttaccgttct gtcgaatggt caagttgtcg
     2581 accaggttgg ggtaacggac tttacggcgg ataagacttc ggctaaagcg gataacgccg
     2641 ataccattac ttataccgcg acggtgaaaa agaatggggt agctcaggct aatgtccctg
     2701 tttcatttaa tattgtttca ggaactgcaa ctcttggggc aaatagtgcc aaaacggatg
     2761 ctaacggtaa ggcaaccgta acgttgaagt cgagtacgcc aggacaggtc gtcgtgtctg
     2821 ctaaaaccgc ggagatgact tcagcactta atgccagtgc ggttatattt tttgatggtg
     2881 cgactagaca agggcagctt gttgagagcg gggggggaag cgtacaagcc gggggatcgc
     2941 tgcgcttgtc gtgtgcagct tccggtattg attccagctc atactgtatg ggttggtttc
     3001 gtcagcgtcc gggaaaggag cgcgagggtg tcgcacgtat caacgggttg ggcggcgtta
     3061 aaacagctta cgctgactct gttaaagatc gtttcaccat tagtcgtgat aatgcagaaa
     3121 atactgttta tttgcaaatg aactccttga aacccgaaga tacagctatt tattattgtg
     3181 ctgccaagtt tagtccaggt tattgcggag gatcgtggag caatttcgga tactggggtc
     3241 aaggaacgca agtgaccgta tcttcgtaat aatactagag ccaggcatca aataaaacga
     3301 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     3361 tactagagtc acactggctc accttcgggt gggcctttct gcgtttatat actagtagcg
     3421 gccgctgcag tccggcaaaa aaacgggcaa ggtgtcacca ccctgccctt tttctttaaa
     3481 accgaaaaga ttacttcgcg ttatgcaggc ttcctcgctc actgactcgc tgcgctcggt
     3541 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatctcga ggttacattg
     3601 tcgatctgtt catggtgaac agctttaaat gcaccaaaaa ctcgtaaaag ctctgatgta
     3661 tctatctttt ttacaccgtt ttcatctgtg catatggaca gttttccctt tgatatctaa
     3721 cggtgaacag ttgttctact tttgtttgtt agtcttgatg cttcactgat agatacaaga
     3781 gccataagaa cctcagatcc ttccgtattt agccagtatg ttctctagtg tggttcgttg
     3841 tttttgcgtg agccatgaga acgaaccatt gagatcatgc ttactttgca tgtcactcaa
     3901 aaattttgcc tcaaaactgg tgagctgaat ttttgcagtt aaagcatcgt gtagtgtttt
     3961 tcttagtccg ttacgtaggt aggaatctga tgtaatggtt gttggtattt tgtcaccatt
     4021 catttttatc tggttgttct caagttcggt tacgagatcc atttgtctat ctagttcaac
     4081 ttggaaaatc aacgtatcag tcgggcggcc tcgcttatca accaccaatt tcatattgct
     4141 gtaagtgttt aaatctttac ttattggttt caaaacccat tggttaagcc ttttaaactc
     4201 atggtagtta ttttcaagca ttaacatgaa cttaaattca tcaaggctaa tctctatatt
     4261 tgccttgtga gttttctttt gtgttagttc ttttaataac cactcataaa tcctcataga
     4321 gtatttgttt tcaaaagact taacatgttc cagattatat tttatgaatt tttttaactg
     4381 gaaaagataa ggcaatatct cttcactaaa aactaattct aatttttcgc ttgagaactt
     4441 ggcatagttt gtccactgga aaatctcaaa gcctttaacc aaaggattcc tgatttccac
     4501 agttctcgtc atcagctctc tggttgcttt agctaataca ccataagcat tttccctact
     4561 gatgttcatc atctgagcgt attggttata agtgaacgat accgtccgtt ctttccttgt
     4621 agggttttca atcgtggggt tgagtagtgc cacacagcat aaaattagct tggtttcatg
     4681 ctccgttaag tcatagcgac taatcgctag ttcatttgct ttgaaaacaa ctaattcaga
     4741 catacatctc aattggtcta ggtgatttta atcactatac caattgagat gggctagtca
     4801 atgataatta catgtccttt tcctttgagt tgtgggtatc tgtaaattct gctagacctt
     4861 tgctggaaaa cttgtaaatt ctgctagacc ctctgtaaat tccgctagac ctttgtgtgt
     4921 tttttttgtt tatattcaag tggttataat ttatagaata aagaaagaat aaaaaaagat
     4981 aaaaagaata gatcccagcc ctgtgtataa ctcactactt tagtcagttc cgcagtatta
     5041 caaaaggatg tcgcaaacgc tgtttgctcc tctacaaaac agaccttaaa accctaaagg
     5101 cttaagtagc accctcgcaa gctcgggcaa atcgctgaat attccttttg tctccgacca
     5161 tcaggcacct gagtcgctgt ctttttcgtg acattcagtt cgctgcgctc acggctctgg
     5221 cagtgaatgg gggtaaatgg cactacaggc gccttttatg gattcatgca aggaaactac
     5281 ccataataca agaaaagccc gtcacgggct tctcagggcg ttttatggcg ggtctgctat
     5341 gtggtgctat ctgacttttt gctgttcagc agttcctgcc ctctgatttt ccagtctgac
     5401 cacttcggat tatcccgtga caggtcattc agactggcta atgcacccag taaggcagcg
     5461 gtatcatcaa caggcttacc cgtcttactg tccctagtgc ttggattctc accaataaaa
     5521 aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact
     5581 ggatctatca acaggagtcc aagcgagctc gtaaacttgg tctgacagtt accaatgctt
     5641 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5701 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5761 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5821 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5881 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5941 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6001 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6061 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6121 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6181 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6241 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6301 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6361 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6421 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6481 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6541 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6601 tcccatggtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     6661 taggcgtatc acgaggcaga atttcagata aaaaaaatcc ttagctttcg ctaaggatga
     6721 tttctggaat tcgcggccgc ttctagag
//
