LOCUS       Exported                6701 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDSG290, complete sequence.
ACCESSION   MH492375
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6701
                     /organism="Cloning vector pDSG290"
                     /mol_type="other DNA"
                     /db_xref="taxon:2268482"
     misc_feature    1..689
                     /label=BBa_I732084
                     /note="BBa_I732084"
     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..686
                     /codon_start=1
                     /product="TetR"
                     /label=TetR
                     /note="BBa_I732080"
                     /protein_id="AXC07644.1"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     regulatory      698..826
                     /regulatory_class="terminator"
                     /note="double terminator (BBa_B0015)"
     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"
     misc_feature    835..896
                     /label=BBa_J13002
                     /note="BBa_J13002"
     misc_feature    835..888
                     /label=pTet (BBa_R0040)
                     /note="pTet (BBa_R0040)"
     protein_bind    835..853
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /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"
     regulatory      3236..3364
                     /regulatory_class="terminator"
                     /note="terminator B0015"
     terminator      3244..3315
                     /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="biobrick suffix"
     regulatory      3386..3457
                     /regulatory_class="terminator"
                     /note="his operon terminator"
     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(3746..4696)
                     /codon_start=1
                     /product="repA"
                     /label=repA
                     /protein_id="AXC07645.1"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     misc_feature    complement(4700..4712)
                     /label=RBS for repA
                     /note="RBS for repA"
     regulatory      complement(4717..4736)
                     /regulatory_class="promoter"
                     /note="repA promoter"
     rep_origin      4744..4966
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      5458..5552
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5582..6442)
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="AXC07646.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6443..6547)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(6443..6521)
                     /regulatory_class="promoter"
                     /note="promoter/RBS for AmpR"
     misc_feature    6562..6582
                     /label=VF2 primer site
                     /note="VF2 primer site"
     terminator      6634..6677
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    6680..6701
                     /label=biobricks prefix
                     /note="biobricks prefix"