LOCUS       40924_15330        5993 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDSG364, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5993)
  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 5993)
  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 5993)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5993)
  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..5993
                     /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..3242
                     /codon_start=1
                     /product="mutated Neae"
                     /label=mutated Neae
                     /note="removed restriction sites"
                     /protein_id="AXC07728.1"
                     /translation="MITHGCYTRTRHKHKLKKTLIMLSAGLGLFFYVNQNSFANGENYF
                     KLGSDSKLLTHDSYQNRLFYTLKTGETVADLSKSQDINLSTIWSLNKHLYSSESEMMKA
                     APGQQIILPLKKLPFEYSALPLLGSAPLVAAGGVAGHTNKLTKMSPDVTKSNMTDDKAL
                     NYAAQQAASLGSQLQSRSLNGDYAKDTALGIAGNQASSQLQAWLQHYGTAEVNLQSGNN
                     FDGSSLDFLLPFYDSEKMLAFGQVGARYIDSRFTANLGAGQRFFLPANMLGYNVFIDQD
                     FSGDNTRLGIGGEYWRDYFKSSVNGYFRMSGWHESYNKKDYDERPANGFDIRFNGYLPS
                     YPALGAKLIYEQYYGDNVALFNSDKLQSNPGAATVGVNYTPIPLVTMGIDYRHGTGNEN
                     DLLYSMQFRYQFDKSWSQQIEPQYVNELRTLSGSRYDLVQRNNNIILEYKKQDILSLNI
                     PHDINGTEHSTQKIQLIVKSKYGLDRIVWDDSALRSQGGQIQHSGSQSAQDYQAILPAY
                     VQGGSNIYKVTARAYDRNGNSSNNVQLTITVLSNGQVVDQVGVTDFTADKTSAKADNAD
                     TITYTATVKKNGVAQANVPVSFNIVSGTATLGANSAKTDANGKATVTLKSSTPGQVVVS
                     AKTAEMTSALNASAVIFFDGATRQVQLQESGGGLVQAGGSLRLSCAASGFSVGNYDMSW
                     IRQARGKGPEGVSGIVSGGVTTWYADSVKGRFTISRDNAENMLYLQMNNLKPEDTAVYY
                     CAPASGHLRGVRGLGTQVTVSS"
     CDS             2889..3242
                     /codon_start=1
                     /product="N2-1_antiAkt1PHMUT-R2AKM3"
                     /label=N2-1_antiAkt1PHMUT-R2AKM3
                     /protein_id="AXC07726.1"
                     /translation="QVQLQESGGGLVQAGGSLRLSCAASGFSVGNYDMSWIRQARGKGP
                     EGVSGIVSGGVTTWYADSVKGRFTISRDNAENMLYLQMNNLKPEDTAVYYCAPASGHLR
                     GVRGLGTQVTVSS"
     misc_feature    3240..3245
                     /label=stop codons
                     /note="stop codons"
     misc_feature    3246..3266
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3267..3324
                     /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(3402..3419)
                     /label=VR primer site
                     /note="VR primer site"
     CDS             complement(3485..4297)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(4892..5436)
                     /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    5854..5874
                     /label=VF2 primer site
                     /note="VF2 primer site"
     terminator      complement(5926..5969)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    5972..5993
                     /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 ggttcaatta caagaatctg gtggtggact ggtgcaagca ggaggttctt
     2941 tacgtctgtc ttgtgcagca tcaggatttt ctgttggtaa ctatgatatg tcgtggatcc
     3001 gtcaggcgcg tggcaaaggt cccgaaggcg tctcgggcat tgttagcgga ggggtaacta
     3061 catggtacgc ggatagcgta aaaggacgtt tcacgattag tcgtgataat gcagagaaca
     3121 tgctttattt gcaaatgaac aaccttaaac ccgaggatac tgctgtctat tattgtgctc
     3181 ctgcgagcgg acatctgcgt ggtgtgcgcg gtctgggtac tcaggtgaca gtgtcaagtt
     3241 aataatacta gtagcggccg ctgcagtccg gcaaaaaagg gcaaggtgtc accaccctgc
     3301 cctttttctt taaaaccgaa aagattactt cgcgttatgc aggcttcctc gctcactgac
     3361 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaatc
     3421 tcgagtcccg tcaagtcagc gtaatgctct gccagtgtta caaccaatta accaattctg
     3481 attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     3541 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     3601 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     3661 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     3721 ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt tcaacaggcc
     3781 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     3841 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
     3901 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     3961 attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat
     4021 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     4081 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     4141 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga
     4201 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     4261 gcctcgagca agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta
     4321 tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc
     4381 agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct gagttgaagg
     4441 atcagatcac gcatcttccc gacaacgcag accgttccgt ggcaaagcaa aagttcaaaa
     4501 tcaccaactg gtccacctac aacaaagctc tcatcaaccg tggctccctc actttctggc
     4561 tggatgatgg ggcgattcag gcctggtatg agtcagcaac accttcttca cgaggcagac
     4621 ctcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg gtgtcagtga
     4681 agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac
     4741 aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga
     4801 gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac
     4861 agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc
     4921 atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc
     4981 aggcgtttcc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg
     5041 tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt
     5101 aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg
     5161 ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg
     5221 cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc
     5281 taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa
     5341 gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca
     5401 gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat taaggggtct
     5461 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5521 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5581 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5641 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5701 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5761 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5821 actttatccg cctccatcca gtctattcca tggtgccacc tgacgtctaa gaaaccatta
     5881 ttatcatgac attaacctat aaaaataggc gtatcacgag gcagaatttc agataaaaaa
     5941 aatccttagc tttcgctaag gatgatttct ggaattcgcg gccgcttcta gag
//
