LOCUS       40924_15220        6382 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDSG287, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6382)
  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 6382)
  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 6382)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6382)
  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..6382
                     /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..2903
                     /codon_start=1
                     /product="mutated Neae"
                     /label=mutated Neae
                     /note="removed restriction sites"
                     /protein_id="AXC07661.1"
                     /translation="MITHGCYTRTRHKHKLKKTLIMLSAGLGLFFYVNQNSFANGENYF
                     KLGSDSKLLTHDSYQNRLFYTLKTGETVADLSKSQDINLSTIWSLNKHLYSSESEMMKA
                     APGQQIILPLKKLPFEYSALPLLGSAPLVAAGGVAGHTNKLTKMSPDVTKSNMTDDKAL
                     NYAAQQAASLGSQLQSRSLNGDYAKDTALGIAGNQASSQLQAWLQHYGTAEVNLQSGNN
                     FDGSSLDFLLPFYDSEKMLAFGQVGARYIDSRFTANLGAGQRFFLPANMLGYNVFIDQD
                     FSGDNTRLGIGGEYWRDYFKSSVNGYFRMSGWHESYNKKDYDERPANGFDIRFNGYLPS
                     YPALGAKLIYEQYYGDNVALFNSDKLQSNPGAATVGVNYTPIPLVTMGIDYRHGTGNEN
                     DLLYSMQFRYQFDKSWSQQIEPQYVNELRTLSGSRYDLVQRNNNIILEYKKQDILSLNI
                     PHDINGTEHSTQKIQLIVKSKYGLDRIVWDDSALRSQGGQIQHSGSQSAQDYQAILPAY
                     VQGGSNIYKVTARAYDRNGNSSNNVQLTITVLSNGQVVDQVGVTDFTADKTSAKADNAD
                     TITYTATVKKNGVAQANVPVSFNIVSGTATLGANSAKTDANGKATVTLKSSTPGQVVVS
                     AKTAEMTSALNASAVIFFDGATREPEA"
     CDS             2889..2903
                     /codon_start=1
                     /product="EPEA peptide protein"
                     /label=EPEA peptide protein
                     /note="C-terminus of alpha synuclein"
                     /protein_id="AXC07662.1"
                     /translation="EPEA"
     misc_feature    2901..2906
                     /label=stop codons
                     /note="stop codons"
     terminator      complement(2926..2969)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3010..3037
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3044..3064
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3065..3124
                     /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(3202..3219)
                     /label=VR primer site
                     /note="VR primer site"
     CDS             complement(3430..4377)
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
     misc_feature    complement(4381..4393)
                     /label=RBS for repA
                     /note="RBS for repA"
     regulatory      complement(4398..4417)
                     /label=repA promoter
                     /note="repA promoter"
                     /regulatory_class="promoter"
     rep_origin      complement(4425..4647)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5139..5233)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5266..6123)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6124..6228)
                     /label=AmpR promoter
     misc_feature    6243..6263
                     /label=VF2 primer site
                     /note="VF2 primer site"
     terminator      complement(6315..6358)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    6361..6382
                     /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 cgactagaga accagaggct taataatact agagccaggc atcaaataaa acgaaaggct
     2941 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag
     3001 agtcacactg gctcaccttc gggtgggcct ttctgcgttt atatactagt agcggccgct
     3061 gcagtccggc aaaaaaacgg gcaaggtgtc accaccctgc cctttttctt taaaaccgaa
     3121 aagattactt cgcgttatgc aggcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3181 gctgcggcga gcggtatcag ctcactcaaa ggcggtaatc tcgaggttac attgtcgatc
     3241 tgttcatggt gaacagcttt aaatgcacca aaaactcgta aaagctctga tgtatctatc
     3301 ttttttacac cgttttcatc tgtgcatatg gacagttttc cctttgatat ctaacggtga
     3361 acagttgttc tacttttgtt tgttagtctt gatgcttcac tgatagatac aagagccata
     3421 agaacctcag atccttccgt atttagccag tatgttctct agtgtggttc gttgtttttg
     3481 cgtgagccat gagaacgaac cattgagatc atgcttactt tgcatgtcac tcaaaaattt
     3541 tgcctcaaaa ctggtgagct gaatttttgc agttaaagca tcgtgtagtg tttttcttag
     3601 tccgttacgt aggtaggaat ctgatgtaat ggttgttggt attttgtcac cattcatttt
     3661 tatctggttg ttctcaagtt cggttacgag atccatttgt ctatctagtt caacttggaa
     3721 aatcaacgta tcagtcgggc ggcctcgctt atcaaccacc aatttcatat tgctgtaagt
     3781 gtttaaatct ttacttattg gtttcaaaac ccattggtta agccttttaa actcatggta
     3841 gttattttca agcattaaca tgaacttaaa ttcatcaagg ctaatctcta tatttgcctt
     3901 gtgagttttc ttttgtgtta gttcttttaa taaccactca taaatcctca tagagtattt
     3961 gttttcaaaa gacttaacat gttccagatt atattttatg aattttttta actggaaaag
     4021 ataaggcaat atctcttcac taaaaactaa ttctaatttt tcgcttgaga acttggcata
     4081 gtttgtccac tggaaaatct caaagccttt aaccaaagga ttcctgattt ccacagttct
     4141 cgtcatcagc tctctggttg ctttagctaa tacaccataa gcattttccc tactgatgtt
     4201 catcatctga gcgtattggt tataagtgaa cgataccgtc cgttctttcc ttgtagggtt
     4261 ttcaatcgtg gggttgagta gtgccacaca gcataaaatt agcttggttt catgctccgt
     4321 taagtcatag cgactaatcg ctagttcatt tgctttgaaa acaactaatt cagacataca
     4381 tctcaattgg tctaggtgat tttaatcact ataccaattg agatgggcta gtcaatgata
     4441 attacatgtc cttttccttt gagttgtggg tatctgtaaa ttctgctaga cctttgctgg
     4501 aaaacttgta aattctgcta gaccctctgt aaattccgct agacctttgt gtgttttttt
     4561 tgtttatatt caagtggtta taatttatag aataaagaaa gaataaaaaa agataaaaag
     4621 aatagatccc agccctgtgt ataactcact actttagtca gttccgcagt attacaaaag
     4681 gatgtcgcaa acgctgtttg ctcctctaca aaacagacct taaaacccta aaggcttaag
     4741 tagcaccctc gcaagctcgg gcaaatcgct gaatattcct tttgtctccg accatcaggc
     4801 acctgagtcg ctgtcttttt cgtgacattc agttcgctgc gctcacggct ctggcagtga
     4861 atgggggtaa atggcactac aggcgccttt tatggattca tgcaaggaaa ctacccataa
     4921 tacaagaaaa gcccgtcacg ggcttctcag ggcgttttat ggcgggtctg ctatgtggtg
     4981 ctatctgact ttttgctgtt cagcagttcc tgccctctga ttttccagtc tgaccacttc
     5041 ggattatccc gtgacaggtc attcagactg gctaatgcac ccagtaaggc agcggtatca
     5101 tcaacaggct tacccgtctt actgtcccta gtgcttggat tctcaccaat aaaaaacgcc
     5161 cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct
     5221 atcaacagga gtccaagcga gctcgtaaac ttggtctgac agttaccaat gcttaatcag
     5281 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5341 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5401 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5461 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5521 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5581 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5641 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5701 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5761 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5821 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5881 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5941 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6001 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6061 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6121 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6181 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttcccat
     6241 ggtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     6301 tatcacgagg cagaatttca gataaaaaaa atccttagct ttcgctaagg atgatttctg
     6361 gaattcgcgg ccgcttctag ag
//
