LOCUS       40924_15580        5855 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDSG420, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5855)
  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 5855)
  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 5855)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5855)
  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..5855
                     /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..3104
                     /codon_start=1
                     /product="mutated Neae"
                     /label=mutated Neae
                     /note="removed restriction sites"
                     /protein_id="AXC07716.1"
                     /translation="MITHGCYTRTRHKHKLKKTLIMLSAGLGLFFYVNQNSFANGENYF
                     KLGSDSKLLTHDSYQNRLFYTLKTGETVADLSKSQDINLSTIWSLNKHLYSSESEMMKA
                     APGQQIILPLKKLPFEYSALPLLGSAPLVAAGGVAGHTNKLTKMSPDVTKSNMTDDKAL
                     NYAAQQAASLGSQLQSRSLNGDYAKDTALGIAGNQASSQLQAWLQHYGTAEVNLQSGNN
                     FDGSSLDFLLPFYDSEKMLAFGQVGARYIDSRFTANLGAGQRFFLPANMLGYNVFIDQD
                     FSGDNTRLGIGGEYWRDYFKSSVNGYFRMSGWHESYNKKDYDERPANGFDIRFNGYLPS
                     YPALGAKLIYEQYYGDNVALFNSDKLQSNPGAATVGVNYTPIPLVTMGIDYRHGTGNEN
                     DLLYSMQFRYQFDKSWSQQIEPQYVNELRTLSGSRYDLVQRNNNIILEYKKQDILSLNI
                     PHDINGTEHSTQKIQLIVKSKYGLDRIVWDDSALRSQGGQIQHSGSQSAQDYQAILPAY
                     VQGGSNIYKVTARAYDRNGNSSNNVQLTITVLSNGQVVDQVGVTDFTADKTSAKADNAD
                     TITYTATVKKNGVAQANVPVSFNIVSGTATLGANSAKTDANGKATVTLKSSTPGQVVVS
                     AKTAEMTSALNASAVIFFDGATRATEVPVSWESFNNGNCFILDLGNNIHQWCGSNSNRY
                     ERLKATQVSKGIRDNERSGRARVHVSEEGTEPEAM"
     CDS             2889..3104
                     /codon_start=1
                     /product="A5_gelsolinFAF"
                     /label=A5_gelsolinFAF
                     /protein_id="AXC07714.1"
                     /translation="ATEVPVSWESFNNGNCFILDLGNNIHQWCGSNSNRYERLKATQVS
                     KGIRDNERSGRARVHVSEEGTEPEAM"
     misc_feature    3102..3107
                     /label=stop codons
                     /note="stop codons"
     misc_feature    3108..3128
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3129..3186
                     /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(3264..3281)
                     /label=VR primer site
                     /note="VR primer site"
     CDS             complement(3347..4159)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(4754..5298)
                     /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    5716..5736
                     /label=VF2 primer site
                     /note="VF2 primer site"
     terminator      complement(5788..5831)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    5834..5855
                     /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 cgactagagc gacagaggta cccgtatcct gggagtcatt caataatgga aattgcttca
     2941 tccttgattt gggaaataac attcatcagt ggtgtgggtc aaacagcaat cgctacgaac
     3001 gtcttaaagc gacacaggtt tctaagggaa tccgtgacaa cgagcgttcc ggtcgtgccc
     3061 gcgtccacgt aagtgaagag ggcacggagc cagaagcaat gtaataatac tagtagcggc
     3121 cgctgcagtc cggcaaaaaa gggcaaggtg tcaccaccct gccctttttc tttaaaaccg
     3181 aaaagattac ttcgcgttat gcaggcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3241 cggctgcggc gagcggtatc agctcactca aaggcggtaa tctcgagtcc cgtcaagtca
     3301 gcgtaatgct ctgccagtgt tacaaccaat taaccaattc tgattagaaa aactcatcga
     3361 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     3421 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     3481 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     3541 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     3601 gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     3661 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     3721 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     3781 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     3841 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     3901 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     3961 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     4021 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     4081 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgag caagacgttt
     4141 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     4201 ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa
     4261 cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagatc acgcatcttc
     4321 ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac tggtccacct
     4381 acaacaaagc tctcatcaac cgtggctccc tcactttctg gctggatgat ggggcgattc
     4441 aggcctggta tgagtcagca acaccttctt cacgaggcag acctcagcgc tagcggagtg
     4501 tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc atgtggcagg
     4561 agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata ttccgcttcc
     4621 tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa
     4681 cggggcggag atttcctgga agatgccagg aagatactta acagggaagt gagagggccg
     4741 cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct
     4801 caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt cccctggcgg
     4861 ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg
     4921 gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc
     4981 tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc
     5041 gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt
     5101 gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt
     5161 tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga
     5221 accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc
     5281 agaccaaaac gatctcaaga agatcatctt attaaggggt ctgacgctca gtggaacgaa
     5341 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5401 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5461 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5521 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5581 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5641 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5701 cagtctattc catggtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     5761 ataaaaatag gcgtatcacg aggcagaatt tcagataaaa aaaatcctta gctttcgcta
     5821 aggatgattt ctggaattcg cggccgcttc tagag
//
