LOCUS       40924_15275        5635 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDSG323, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5635)
  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 5635)
  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 5635)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5635)
  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..5635
                     /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..2891
                     /codon_start=1
                     /product="mutated Neae"
                     /label=mutated Neae
                     /note="removed restriction sites"
                     /protein_id="AXC07640.1"
                     /translation="MITHGCYTRTRHKHKLKKTLIMLSAGLGLFFYVNQNSFANGENYF
                     KLGSDSKLLTHDSYQNRLFYTLKTGETVADLSKSQDINLSTIWSLNKHLYSSESEMMKA
                     APGQQIILPLKKLPFEYSALPLLGSAPLVAAGGVAGHTNKLTKMSPDVTKSNMTDDKAL
                     NYAAQQAASLGSQLQSRSLNGDYAKDTALGIAGNQASSQLQAWLQHYGTAEVNLQSGNN
                     FDGSSLDFLLPFYDSEKMLAFGQVGARYIDSRFTANLGAGQRFFLPANMLGYNVFIDQD
                     FSGDNTRLGIGGEYWRDYFKSSVNGYFRMSGWHESYNKKDYDERPANGFDIRFNGYLPS
                     YPALGAKLIYEQYYGDNVALFNSDKLQSNPGAATVGVNYTPIPLVTMGIDYRHGTGNEN
                     DLLYSMQFRYQFDKSWSQQIEPQYVNELRTLSGSRYDLVQRNNNIILEYKKQDILSLNI
                     PHDINGTEHSTQKIQLIVKSKYGLDRIVWDDSALRSQGGQIQHSGSQSAQDYQAILPAY
                     VQGGSNIYKVTARAYDRNGNSSNNVQLTITVLSNGQVVDQVGVTDFTADKTSAKADNAD
                     TITYTATVKKNGVAQANVPVSFNIVSGTATLGANSAKTDANGKATVTLKSSTPGQVVVS
                     AKTAEMTSALNASAVIFFDGATS"
     misc_feature    2883..2908
                     /label=RFC23 biobrick suffix
                     /note="RFC23 biobrick suffix"
     terminator      2909..2966
                     /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(3044..3061)
                     /label=VR primer site
                     /note="VR primer site"
     CDS             complement(3127..3939)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(4534..5078)
                     /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    5496..5516
                     /label=VF2 primer site
                     /note="VF2 primer site"
     terminator      complement(5568..5611)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    5614..5635
                     /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 cgactagtta ataaagcggc cgctgcagtc cggcaaaaaa gggcaaggtg tcaccaccct
     2941 gccctttttc tttaaaaccg aaaagattac ttcgcgttat gcaggcttcc tcgctcactg
     3001 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3061 tctcgagtcc cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc
     3121 tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     3181 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     3241 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     3301 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     3361 gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg
     3421 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     3481 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     3541 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     3601 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     3661 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     3721 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     3781 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     3841 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     3901 cggcctcgag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     3961 tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca
     4021 tcagagattt tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa
     4081 ggatcagatc acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa
     4141 aatcaccaac tggtccacct acaacaaagc tctcatcaac cgtggctccc tcactttctg
     4201 gctggatgat ggggcgattc aggcctggta tgagtcagca acaccttctt cacgaggcag
     4261 acctcagcgc tagcggagtg tatactggct tactatgttg gcactgatga gggtgtcagt
     4321 gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat
     4381 acaggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc
     4441 gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta
     4501 acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa
     4561 gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata
     4621 ccaggcgttt cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc
     4681 ggtgtcattc cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagttccgg
     4741 gtaggcagtt cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg
     4801 cgccttatcc ggtaactatc gtcttgagtc caacccggaa agacatgcaa aagcaccact
     4861 ggcagcagcc actggtaatt gatttagagg agttagtctt gaagtcatgc gccggttaag
     4921 gctaaactga aaggacaagt tttggtgact gcgctcctcc aagccagtta cctcggttca
     4981 aagagttggt agctcagaga accttcgaaa aaccgccctg caaggcggtt ttttcgtttt
     5041 cagagcaaga gattacgcgc agaccaaaac gatctcaaga agatcatctt attaaggggt
     5101 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5161 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5221 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5281 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5341 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5401 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5461 caactttatc cgcctccatc cagtctattc catggtgcca cctgacgtct aagaaaccat
     5521 tattatcatg acattaacct ataaaaatag gcgtatcacg aggcagaatt tcagataaaa
     5581 aaaatcctta gctttcgcta aggatgattt ctggaattcg cggccgcttc tagag
//
