LOCUS       40924_15255        5791 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDSG318, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5791)
  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 5791)
  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 5791)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5791)
  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..5791
                     /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)
                     /note="RBS (BBa_B0034)"
     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)(1)
                     /note="RBS (BBa_B0034)(1)"
     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="AXC07695.1"
                     /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="AXC07696.1"
                     /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..3122
                     /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(3200..3217)
                     /label=VR primer site
                     /note="VR primer site"
     CDS             complement(3283..4095)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(4690..5234)
                     /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    5652..5672
                     /label=VF2 primer site
                     /note="VF2 primer site"
     terminator      complement(5724..5767)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    5770..5791
                     /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 aaaaaagggc aaggtgtcac caccctgccc tttttcttta aaaccgaaaa
     3121 gattacttcg cgttatgcag gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3181 tgcggcgagc ggtatcagct cactcaaagg cggtaatctc gagtcccgtc aagtcagcgt
     3241 aatgctctgc cagtgttaca accaattaac caattctgat tagaaaaact catcgagcat
     3301 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
     3361 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
     3421 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
     3481 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
     3541 aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     3601 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac
     3661 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
     3721 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
     3781 tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
     3841 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
     3901 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     3961 cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     4021 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgagcaag acgtttcccg
     4081 ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
     4141 tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg
     4201 gctttgttga ataaatcgaa cttttgctga gttgaaggat cagatcacgc atcttcccga
     4261 caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc accaactggt ccacctacaa
     4321 caaagctctc atcaaccgtg gctccctcac tttctggctg gatgatgggg cgattcaggc
     4381 ctggtatgag tcagcaacac cttcttcacg aggcagacct cagcgctagc ggagtgtata
     4441 ctggcttact atgttggcac tgatgagggt gtcagtgaag tgcttcatgt ggcaggagaa
     4501 aaaaggctgc accggtgcgt cagcagaata tgtgatacag gatatattcc gcttcctcgc
     4561 tcactgactc gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg
     4621 gcggagattt cctggaagat gccaggaaga tacttaacag ggaagtgaga gggccgcggc
     4681 aaagccgttt ttccataggc tccgcccccc tgacaagcat cacgaaatct gacgctcaaa
     4741 tcagtggtgg cgaaacccga caggactata aagataccag gcgtttcccc tggcggctcc
     4801 ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg tcattccgct gttatggccg
     4861 cgtttgtctc attccacgcc tgacactcag ttccgggtag gcagttcgct ccaagctgga
     4921 ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc ttatccggta actatcgtct
     4981 tgagtccaac ccggaaagac atgcaaaagc accactggca gcagccactg gtaattgatt
     5041 tagaggagtt agtcttgaag tcatgcgccg gttaaggcta aactgaaagg acaagttttg
     5101 gtgactgcgc tcctccaagc cagttacctc ggttcaaaga gttggtagct cagagaacct
     5161 tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga gcaagagatt acgcgcagac
     5221 caaaacgatc tcaagaagat catcttatta aggggtctga cgctcagtgg aacgaaaact
     5281 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5341 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5401 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5461 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5521 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5581 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5641 ctattccatg gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     5701 aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt tcgctaagga
     5761 tgatttctgg aattcgcggc cgcttctaga g
//
