LOCUS       40924_15235        6701 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDSG290, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6701)
  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 6701)
  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 6701)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6701)
  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..6701
                     /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..3224
                     /codon_start=1
                     /product="mutated Neae"
                     /label=mutated Neae
                     /note="removed restriction sites"
                     /protein_id="AXC07642.1"
                     /translation="MITHGCYTRTRHKHKLKKTLIMLSAGLGLFFYVNQNSFANGENYF
                     KLGSDSKLLTHDSYQNRLFYTLKTGETVADLSKSQDINLSTIWSLNKHLYSSESEMMKA
                     APGQQIILPLKKLPFEYSALPLLGSAPLVAAGGVAGHTNKLTKMSPDVTKSNMTDDKAL
                     NYAAQQAASLGSQLQSRSLNGDYAKDTALGIAGNQASSQLQAWLQHYGTAEVNLQSGNN
                     FDGSSLDFLLPFYDSEKMLAFGQVGARYIDSRFTANLGAGQRFFLPANMLGYNVFIDQD
                     FSGDNTRLGIGGEYWRDYFKSSVNGYFRMSGWHESYNKKDYDERPANGFDIRFNGYLPS
                     YPALGAKLIYEQYYGDNVALFNSDKLQSNPGAATVGVNYTPIPLVTMGIDYRHGTGNEN
                     DLLYSMQFRYQFDKSWSQQIEPQYVNELRTLSGSRYDLVQRNNNIILEYKKQDILSLNI
                     PHDINGTEHSTQKIQLIVKSKYGLDRIVWDDSALRSQGGQIQHSGSQSAQDYQAILPAY
                     VQGGSNIYKVTARAYDRNGNSSNNVQLTITVLSNGQVVDQVGVTDFTADKTSAKADNAD
                     TITYTATVKKNGVAQANVPVSFNIVSGTATLGANSAKTDANGKATVTLKSSTPGQVVVS
                     AKTAEMTSALNASAVIFFDGATRQVQLQESGGGSVRPGESLRLSCIAAGTLFSGKAVGW
                     YRQAPGKDRELVAALSSGRETYYIGPVKGRFIISRDDNKDTLYLQMNNLKLEDTAIYTC
                     RLGLHWGQGTQVTVSS"
     CDS             2889..3224
                     /codon_start=1
                     /product="human P53 transactivation domain nanobody R4P43"
                     /label=human P53 transactivation domain nanobody R4P43
                     /protein_id="AXC07643.1"
                     /translation="QVQLQESGGGSVRPGESLRLSCIAAGTLFSGKAVGWYRQAPGKDR
                     ELVAALSSGRETYYIGPVKGRFIISRDDNKDTLYLQMNNLKLEDTAIYTCRLGLHWGQG
                     TQVTVSS"
     misc_feature    3222..3227
                     /label=stop codons
                     /note="stop codons"
     terminator      complement(3247..3290)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3331..3358
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3365..3385
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3386..3443
                     /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(3521..3538)
                     /label=VR primer site
                     /note="VR primer site"
     CDS             complement(3749..4696)
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
     misc_feature    complement(4700..4712)
                     /label=RBS for repA
                     /note="RBS for repA"
     regulatory      complement(4717..4736)
                     /label=repA promoter
                     /note="repA promoter"
                     /regulatory_class="promoter"
     rep_origin      complement(4744..4966)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5458..5552)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5585..6442)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6443..6547)
                     /label=AmpR promoter
     misc_feature    6562..6582
                     /label=VF2 primer site
                     /note="VF2 primer site"
     terminator      complement(6634..6677)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    6680..6701
                     /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 ggtgcagcta caggagtctg ggggaggctc ggtgcggcct ggggagtctc
     2941 ttcgactctc ctgtatagcc gctggaacct tgtttagtgg caaagccgtg ggctggtacc
     3001 gccaggctcc agggaaggac cgcgagctgg tcgcagctct tagtagtggt cgtgaaacat
     3061 actatatagg gcctgtgaag ggccgattca ttatctccag agacgataat aaagacacac
     3121 tgtatctgca aatgaacaac ctaaaacttg aggacacggc catctatacc tgtaggttag
     3181 gcctacattg gggccagggg acccaggtca ccgtctcctc ataataatac tagagccagg
     3241 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     3301 tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt
     3361 tatatactag tagcggccgc tgcagtccgg caaaaaaggg caaggtgtca ccaccctgcc
     3421 ctttttcttt aaaaccgaaa agattacttc gcgttatgca ggcttcctcg ctcactgact
     3481 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatct
     3541 cgaggttaca ttgtcgatct gttcatggtg aacagcttta aatgcaccaa aaactcgtaa
     3601 aagctctgat gtatctatct tttttacacc gttttcatct gtgcatatgg acagttttcc
     3661 ctttgatatc taacggtgaa cagttgttct acttttgttt gttagtcttg atgcttcact
     3721 gatagataca agagccataa gaacctcaga tccttccgta tttagccagt atgttctcta
     3781 gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc attgagatca tgcttacttt
     3841 gcatgtcact caaaaatttt gcctcaaaac tggtgagctg aatttttgca gttaaagcat
     3901 cgtgtagtgt ttttcttagt ccgttacgta ggtaggaatc tgatgtaatg gttgttggta
     3961 ttttgtcacc attcattttt atctggttgt tctcaagttc ggttacgaga tccatttgtc
     4021 tatctagttc aacttggaaa atcaacgtat cagtcgggcg gcctcgctta tcaaccacca
     4081 atttcatatt gctgtaagtg tttaaatctt tacttattgg tttcaaaacc cattggttaa
     4141 gccttttaaa ctcatggtag ttattttcaa gcattaacat gaacttaaat tcatcaaggc
     4201 taatctctat atttgccttg tgagttttct tttgtgttag ttcttttaat aaccactcat
     4261 aaatcctcat agagtatttg ttttcaaaag acttaacatg ttccagatta tattttatga
     4321 atttttttaa ctggaaaaga taaggcaata tctcttcact aaaaactaat tctaattttt
     4381 cgcttgagaa cttggcatag tttgtccact ggaaaatctc aaagccttta accaaaggat
     4441 tcctgatttc cacagttctc gtcatcagct ctctggttgc tttagctaat acaccataag
     4501 cattttccct actgatgttc atcatctgag cgtattggtt ataagtgaac gataccgtcc
     4561 gttctttcct tgtagggttt tcaatcgtgg ggttgagtag tgccacacag cataaaatta
     4621 gcttggtttc atgctccgtt aagtcatagc gactaatcgc tagttcattt gctttgaaaa
     4681 caactaattc agacatacat ctcaattggt ctaggtgatt ttaatcacta taccaattga
     4741 gatgggctag tcaatgataa ttacatgtcc ttttcctttg agttgtgggt atctgtaaat
     4801 tctgctagac ctttgctgga aaacttgtaa attctgctag accctctgta aattccgcta
     4861 gacctttgtg tgtttttttt gtttatattc aagtggttat aatttataga ataaagaaag
     4921 aataaaaaaa gataaaaaga atagatccca gccctgtgta taactcacta ctttagtcag
     4981 ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt
     5041 aaaaccctaa aggcttaagt agcaccctcg caagctcggg caaatcgctg aatattcctt
     5101 ttgtctccga ccatcaggca cctgagtcgc tgtctttttc gtgacattca gttcgctgcg
     5161 ctcacggctc tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt atggattcat
     5221 gcaaggaaac tacccataat acaagaaaag cccgtcacgg gcttctcagg gcgttttatg
     5281 gcgggtctgc tatgtggtgc tatctgactt tttgctgttc agcagttcct gccctctgat
     5341 tttccagtct gaccacttcg gattatcccg tgacaggtca ttcagactgg ctaatgcacc
     5401 cagtaaggca gcggtatcat caacaggctt acccgtctta ctgtccctag tgcttggatt
     5461 ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc
     5521 tgaggtcatt actggatcta tcaacaggag tccaagcgag ctcgtaaact tggtctgaca
     5581 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5641 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5701 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5761 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5821 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5881 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5941 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6001 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6061 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6121 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6181 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6241 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6301 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6361 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6421 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6481 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6541 ttccgcgcac atttcccatg gtgccacctg acgtctaaga aaccattatt atcatgacat
     6601 taacctataa aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt
     6661 tcgctaagga tgatttctgg aattcgcggc cgcttctaga g
//
