LOCUS       40924_15380        6020 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDSG374, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6020)
  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 6020)
  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 6020)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6020)
  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..6020
                     /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..3269
                     /codon_start=1
                     /product="mutated Neae"
                     /label=mutated Neae
                     /note="removed restriction sites"
                     /protein_id="AXC07740.1"
                     /translation="MITHGCYTRTRHKHKLKKTLIMLSAGLGLFFYVNQNSFANGENYF
                     KLGSDSKLLTHDSYQNRLFYTLKTGETVADLSKSQDINLSTIWSLNKHLYSSESEMMKA
                     APGQQIILPLKKLPFEYSALPLLGSAPLVAAGGVAGHTNKLTKMSPDVTKSNMTDDKAL
                     NYAAQQAASLGSQLQSRSLNGDYAKDTALGIAGNQASSQLQAWLQHYGTAEVNLQSGNN
                     FDGSSLDFLLPFYDSEKMLAFGQVGARYIDSRFTANLGAGQRFFLPANMLGYNVFIDQD
                     FSGDNTRLGIGGEYWRDYFKSSVNGYFRMSGWHESYNKKDYDERPANGFDIRFNGYLPS
                     YPALGAKLIYEQYYGDNVALFNSDKLQSNPGAATVGVNYTPIPLVTMGIDYRHGTGNEN
                     DLLYSMQFRYQFDKSWSQQIEPQYVNELRTLSGSRYDLVQRNNNIILEYKKQDILSLNI
                     PHDINGTEHSTQKIQLIVKSKYGLDRIVWDDSALRSQGGQIQHSGSQSAQDYQAILPAY
                     VQGGSNIYKVTARAYDRNGNSSNNVQLTITVLSNGQVVDQVGVTDFTADKTSAKADNAD
                     TITYTATVKKNGVAQANVPVSFNIVSGTATLGANSAKTDANGKATVTLKSSTPGQVVVS
                     AKTAEMTSALNASAVIFFDGATRQVQLQESGGGLVQAGGSLRLSCAASGRTVSSYAMGW
                     FRQAPGKEREFVAAISWSGSTTYYSDSVKGRFTFSRDKPKNTVYLQMNSLKPEDTAVYY
                     CAASRIITSQPMTTMGEYHYWGQGTQVTVSS"
     CDS             2889..3269
                     /codon_start=1
                     /product="N3-3_antiAkt3PH-4AKH8"
                     /label=N3-3_antiAkt3PH-4AKH8
                     /protein_id="AXC07738.1"
                     /translation="QVQLQESGGGLVQAGGSLRLSCAASGRTVSSYAMGWFRQAPGKER
                     EFVAAISWSGSTTYYSDSVKGRFTFSRDKPKNTVYLQMNSLKPEDTAVYYCAASRIITS
                     QPMTTMGEYHYWGQGTQVTVSS"
     misc_feature    3267..3272
                     /label=stop codons
                     /note="stop codons"
     misc_feature    3273..3293
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3294..3351
                     /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(3429..3446)
                     /label=VR primer site
                     /note="VR primer site"
     CDS             complement(3512..4324)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(4919..5463)
                     /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    5881..5901
                     /label=VF2 primer site
                     /note="VF2 primer site"
     terminator      complement(5953..5996)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    5999..6020
                     /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 agtccaactt caagagagcg ggggtggttt agtgcaggcc ggggggtcgc
     2941 ttcgcttgtc atgtgctgca agcgggcgca cagtttcatc ttatgctatg ggttggttcc
     3001 gccaagctcc tggtaaggaa cgtgagtttg tagccgccat ctcatggagt ggatccacaa
     3061 cctactatag cgattcagtg aagggccgct tcacattttc gcgcgataaa ccgaagaaca
     3121 ctgtatactt gcaaatgaac agtctgaaac ctgaggacac agccgtttac tactgcgctg
     3181 ccagtcgcat tattacatcg cagccaatga caacgatggg ggaatatcat tactgggggc
     3241 agggtactca ggtgacggtt tcgtcttaat aatactagta gcggccgctg cagtccggca
     3301 aaaaagggca aggtgtcacc accctgccct ttttctttaa aaccgaaaag attacttcgc
     3361 gttatgcagg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3421 gtatcagctc actcaaaggc ggtaatctcg agtcccgtca agtcagcgta atgctctgcc
     3481 agtgttacaa ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact
     3541 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     3601 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     3661 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     3721 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca
     3781 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     3841 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt
     3901 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     3961 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg
     4021 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     4081 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     4141 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     4201 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     4261 cagcatccat gttggaattt aatcgcggcc tcgagcaaga cgtttcccgt tgaatatggc
     4321 tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata
     4381 tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa
     4441 taaatcgaac ttttgctgag ttgaaggatc agatcacgca tcttcccgac aacgcagacc
     4501 gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc cacctacaac aaagctctca
     4561 tcaaccgtgg ctccctcact ttctggctgg atgatggggc gattcaggcc tggtatgagt
     4621 cagcaacacc ttcttcacga ggcagacctc agcgctagcg gagtgtatac tggcttacta
     4681 tgttggcact gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca
     4741 ccggtgcgtc agcagaatat gtgatacagg atatattccg cttcctcgct cactgactcg
     4801 ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc
     4861 ctggaagatg ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt
     4921 tccataggct ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc
     4981 gaaacccgac aggactataa agataccagg cgtttcccct ggcggctccc tcgtgcgctc
     5041 tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca
     5101 ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg
     5161 aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5221 cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta
     5281 gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct
     5341 cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg
     5401 ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct
     5461 caagaagatc atcttattaa ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5521 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5581 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5641 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5701 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5761 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5821 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattccatgg
     5881 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     5941 tcacgaggca gaatttcaga taaaaaaaat ccttagcttt cgctaaggat gatttctgga
     6001 attcgcggcc gcttctagag
//
