LOCUS       40924_15375        5981 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDSG373, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5981)
  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 5981)
  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 5981)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5981)
  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..5981
                     /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..3230
                     /codon_start=1
                     /product="mutated Neae"
                     /label=mutated Neae
                     /note="removed restriction sites"
                     /protein_id="AXC07748.1"
                     /translation="MITHGCYTRTRHKHKLKKTLIMLSAGLGLFFYVNQNSFANGENYF
                     KLGSDSKLLTHDSYQNRLFYTLKTGETVADLSKSQDINLSTIWSLNKHLYSSESEMMKA
                     APGQQIILPLKKLPFEYSALPLLGSAPLVAAGGVAGHTNKLTKMSPDVTKSNMTDDKAL
                     NYAAQQAASLGSQLQSRSLNGDYAKDTALGIAGNQASSQLQAWLQHYGTAEVNLQSGNN
                     FDGSSLDFLLPFYDSEKMLAFGQVGARYIDSRFTANLGAGQRFFLPANMLGYNVFIDQD
                     FSGDNTRLGIGGEYWRDYFKSSVNGYFRMSGWHESYNKKDYDERPANGFDIRFNGYLPS
                     YPALGAKLIYEQYYGDNVALFNSDKLQSNPGAATVGVNYTPIPLVTMGIDYRHGTGNEN
                     DLLYSMQFRYQFDKSWSQQIEPQYVNELRTLSGSRYDLVQRNNNIILEYKKQDILSLNI
                     PHDINGTEHSTQKIQLIVKSKYGLDRIVWDDSALRSQGGQIQHSGSQSAQDYQAILPAY
                     VQGGSNIYKVTARAYDRNGNSSNNVQLTITVLSNGQVVDQVGVTDFTADKTSAKADNAD
                     TITYTATVKKNGVAQANVPVSFNIVSGTATLGANSAKTDANGKATVTLKSSTPGQVVVS
                     AKTAEMTSALNASAVIFFDGATRQVQLQESGGGLVQPGGSLRLSCAASGFTFSKYPMSW
                     VRQAPGKGLEWISTITSTGASTTYADSVSGRFTISRDNAKNTLYLQMNSLKPEDTAVYY
                     CRNSRGSRGQGTQVTVSS"
     CDS             2889..3230
                     /codon_start=1
                     /product="N3-2_antiAkt3PH-3AKH59"
                     /label=N3-2_antiAkt3PH-3AKH59
                     /protein_id="AXC07746.1"
                     /translation="QVQLQESGGGLVQPGGSLRLSCAASGFTFSKYPMSWVRQAPGKGL
                     EWISTITSTGASTTYADSVSGRFTISRDNAKNTLYLQMNSLKPEDTAVYYCRNSRGSRG
                     QGTQVTVSS"
     misc_feature    3228..3233
                     /label=stop codons
                     /note="stop codons"
     misc_feature    3234..3254
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3255..3312
                     /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(3390..3407)
                     /label=VR primer site
                     /note="VR primer site"
     CDS             complement(3473..4285)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(4880..5424)
                     /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    5842..5862
                     /label=VF2 primer site
                     /note="VF2 primer site"
     terminator      complement(5914..5957)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    5960..5981
                     /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 caagagtcag gcggtggctt ggtccagccc ggcggctccc
     2941 tgcgcttgag ctgtgcggca tcaggtttca catttagtaa atacccaatg tcatgggttc
     3001 gtcaagctcc gggtaaaggt ttagaatgga ttagtacgat cacttccacg ggtgcatcaa
     3061 ccacatatgc tgactctgta tctggacgct ttacgatctc tcgcgataat gccaagaaca
     3121 ctttatatct gcaaatgaac agccttaaac ctgaagacac agcggtctac tactgtcgta
     3181 acagtcgtgg ttctcgtgga cagggcacgc aggtgactgt gagttcttaa taatactagt
     3241 agcggccgct gcagtccggc aaaaaagggc aaggtgtcac caccctgccc tttttcttta
     3301 aaaccgaaaa gattacttcg cgttatgcag gcttcctcgc tcactgactc gctgcgctcg
     3361 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatctc gagtcccgtc
     3421 aagtcagcgt aatgctctgc cagtgttaca accaattaac caattctgat tagaaaaact
     3481 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     3541 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     3601 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     3661 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     3721 agaatggcaa aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     3781 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     3841 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     3901 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     3961 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     4021 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     4081 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     4141 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     4201 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgagcaag
     4261 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
     4321 gttttattgt tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag
     4381 acacaacgtg gctttgttga ataaatcgaa cttttgctga gttgaaggat cagatcacgc
     4441 atcttcccga caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc accaactggt
     4501 ccacctacaa caaagctctc atcaaccgtg gctccctcac tttctggctg gatgatgggg
     4561 cgattcaggc ctggtatgag tcagcaacac cttcttcacg aggcagacct cagcgctagc
     4621 ggagtgtata ctggcttact atgttggcac tgatgagggt gtcagtgaag tgcttcatgt
     4681 ggcaggagaa aaaaggctgc accggtgcgt cagcagaata tgtgatacag gatatattcc
     4741 gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct
     4801 tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag ggaagtgaga
     4861 gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat cacgaaatct
     4921 gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4981 tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg tcattccgct
     5041 gttatggccg cgtttgtctc attccacgcc tgacactcag ttccgggtag gcagttcgct
     5101 ccaagctgga ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc ttatccggta
     5161 actatcgtct tgagtccaac ccggaaagac atgcaaaagc accactggca gcagccactg
     5221 gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta aactgaaagg
     5281 acaagttttg gtgactgcgc tcctccaagc cagttacctc ggttcaaaga gttggtagct
     5341 cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga gcaagagatt
     5401 acgcgcagac caaaacgatc tcaagaagat catcttatta aggggtctga cgctcagtgg
     5461 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5521 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5581 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5641 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5701 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5761 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5821 tccatccagt ctattccatg gtgccacctg acgtctaaga aaccattatt atcatgacat
     5881 taacctataa aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt
     5941 tcgctaagga tgatttctgg aattcgcggc cgcttctaga g
//
