LOCUS       40924_15505        5999 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDSG399, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5999)
  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 5999)
  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 5999)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5999)
  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..5999
                     /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..3248
                     /codon_start=1
                     /product="mutated Neae"
                     /label=mutated Neae
                     /note="removed restriction sites"
                     /protein_id="AXC07944.1"
                     /translation="MITHGCYTRTRHKHKLKKTLIMLSAGLGLFFYVNQNSFANGENYF
                     KLGSDSKLLTHDSYQNRLFYTLKTGETVADLSKSQDINLSTIWSLNKHLYSSESEMMKA
                     APGQQIILPLKKLPFEYSALPLLGSAPLVAAGGVAGHTNKLTKMSPDVTKSNMTDDKAL
                     NYAAQQAASLGSQLQSRSLNGDYAKDTALGIAGNQASSQLQAWLQHYGTAEVNLQSGNN
                     FDGSSLDFLLPFYDSEKMLAFGQVGARYIDSRFTANLGAGQRFFLPANMLGYNVFIDQD
                     FSGDNTRLGIGGEYWRDYFKSSVNGYFRMSGWHESYNKKDYDERPANGFDIRFNGYLPS
                     YPALGAKLIYEQYYGDNVALFNSDKLQSNPGAATVGVNYTPIPLVTMGIDYRHGTGNEN
                     DLLYSMQFRYQFDKSWSQQIEPQYVNELRTLSGSRYDLVQRNNNIILEYKKQDILSLNI
                     PHDINGTEHSTQKIQLIVKSKYGLDRIVWDDSALRSQGGQIQHSGSQSAQDYQAILPAY
                     VQGGSNIYKVTARAYDRNGNSSNNVQLTITVLSNGQVVDQVGVTDFTADKTSAKADNAD
                     TITYTATVKKNGVAQANVPVSFNIVSGTATLGANSAKTDANGKATVTLKSSTPGQVVVS
                     AKTAEMTSALNASAVIFFDGATRQVQLQESGGGLVQPGGSLRLSCAASGIIFSDRAVGW
                     YRQAAGKQRELVAAATSGGNIKYADFVKGRFTISRDDAKNTVYLQMNSLKPEDTAVYYC
                     RVGRHIRQYGYDYWGAGTQVTVSS"
     CDS             2889..3248
                     /codon_start=1
                     /product="N8-2_antiP53TA-R3P12"
                     /label=N8-2_antiP53TA-R3P12
                     /protein_id="AXC07942.1"
                     /translation="QVQLQESGGGLVQPGGSLRLSCAASGIIFSDRAVGWYRQAAGKQR
                     ELVAAATSGGNIKYADFVKGRFTISRDDAKNTVYLQMNSLKPEDTAVYYCRVGRHIRQY
                     GYDYWGAGTQVTVSS"
     misc_feature    3246..3251
                     /label=stop codons
                     /note="stop codons"
     misc_feature    3252..3272
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3273..3330
                     /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(3408..3425)
                     /label=VR primer site
                     /note="VR primer site"
     CDS             complement(3491..4303)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(4898..5442)
                     /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    5860..5880
                     /label=VF2 primer site
                     /note="VF2 primer site"
     terminator      complement(5932..5975)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    5978..5999
                     /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 ggtgcaactt caggagagcg gaggcgggct ggtacaacca ggaggatcct
     2941 tgcgtttatc atgcgcggcc tccggtatca tcttttctga tcgcgctgtg ggctggtatc
     3001 gccaagcagc tggcaagcaa cgtgagcttg ttgcggcagc tacaagcggg gggaacatca
     3061 agtacgcgga ttttgtcaaa ggacgtttca cgatttcccg cgacgacgcg aaaaacactg
     3121 tctacttaca gatgaatagt cttaaacctg aagatacagc tgtatattac tgccgtgtcg
     3181 gtcgtcatat ccgccaatat ggttatgact actggggagc aggtacacag gtcaccgttt
     3241 cctcataata atactagtag cggccgctgc agtccggcaa aaaagggcaa ggtgtcacca
     3301 ccctgccctt tttctttaaa accgaaaaga ttacttcgcg ttatgcaggc ttcctcgctc
     3361 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3421 gtaatctcga gtcccgtcaa gtcagcgtaa tgctctgcca gtgttacaac caattaacca
     3481 attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
     3541 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     3601 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     3661 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     3721 tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag acttgttcaa
     3781 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     3841 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     3901 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     3961 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
     4021 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     4081 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     4141 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     4201 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     4261 atcgcggcct cgagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac
     4321 tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt
     4381 aacatcagag attttgagac acaacgtggc tttgttgaat aaatcgaact tttgctgagt
     4441 tgaaggatca gatcacgcat cttcccgaca acgcagaccg ttccgtggca aagcaaaagt
     4501 tcaaaatcac caactggtcc acctacaaca aagctctcat caaccgtggc tccctcactt
     4561 tctggctgga tgatggggcg attcaggcct ggtatgagtc agcaacacct tcttcacgag
     4621 gcagacctca gcgctagcgg agtgtatact ggcttactat gttggcactg atgagggtgt
     4681 cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac cggtgcgtca gcagaatatg
     4741 tgatacagga tatattccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg
     4801 cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc caggaagata
     4861 cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc cgcccccctg
     4921 acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca ggactataaa
     4981 gataccaggc gtttcccctg gcggctccct cgtgcgctct cctgttcctg cctttcggtt
     5041 taccggtgtc attccgctgt tatggccgcg tttgtctcat tccacgcctg acactcagtt
     5101 ccgggtaggc agttcgctcc aagctggact gtatgcacga accccccgtt cagtccgacc
     5161 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac
     5221 cactggcagc agccactggt aattgattta gaggagttag tcttgaagtc atgcgccggt
     5281 taaggctaaa ctgaaaggac aagttttggt gactgcgctc ctccaagcca gttacctcgg
     5341 ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg
     5401 ttttcagagc aagagattac gcgcagacca aaacgatctc aagaagatca tcttattaag
     5461 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5521 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5581 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5641 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5701 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5761 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5821 cctgcaactt tatccgcctc catccagtct attccatggt gccacctgac gtctaagaaa
     5881 ccattattat catgacatta acctataaaa ataggcgtat cacgaggcag aatttcagat
     5941 aaaaaaaatc cttagctttc gctaaggatg atttctggaa ttcgcggccg cttctagag
//
