LOCUS       40924_630        5007 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector DH6-78, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5007)
  AUTHORS   Huang DC, Holtz WJ, Maharbiz MM.
  TITLE     A genetic bistable switch utilizing nonlinear protein degradation
  JOURNAL   J Biol Eng 6 (1), 9 (2012)
  PUBMED    22776405
REFERENCE   2  (bases 1 to 5007)
  AUTHORS   Huang DC, Holtz WJ, Maharbiz MM.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-JUN-2012) Electrical Engineering and Computer 
            Sciences, University of California, Berkeley, 656 Sutardja Dai Hall,
            Berkeley, CA 94720, USA
REFERENCE   3  (bases 1 to 5007)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5007)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Eng"; date: "2012"; volume: "6"; issue: "1"; pages: "9"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-JUN-2012) Electrical Engineering and Computer Sciences, 
            University of California, Berkeley, 656 Sutardja Dai Hall, Berkeley,
            CA 94720, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: GENtle v. 1.9.4
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..82
                     /label=pRM
                     /note="pRM"
                     /regulatory_class="promoter"
     misc_feature    9..26
                     /label=oR1
                     /note="oR1"
     misc_feature    33..50
                     /label=oR2
                     /note="oR2"
     misc_feature    59..65
                     /label=oR3mut
                     /note="oR3mut"
     5'UTR           89..123
     CDS             124..834
                     /label=lambda repressor
                     /note="phage lambda repressor"
     5'UTR           847..887
     CDS             888..1508
                     /label=TetR
                     /note="tetracycline repressor TetR"
     CDS             1509..1541
                     /label=ssrA tag (LVA)
                     /note="C-terminal peptide that mediates degradation in
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
     terminator      1580..1651
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1667..1694
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2228..3175)
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
     rep_origin      complement(3223..3445)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3937..4031)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4065..4856)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 gcaaccatta tcaccgccag aggtaaaata gtcaacacgc acggtgttag atatttataa
       61 atagtggtga tagatttaac gtggatctaa tccaacaata tagcatactc tcgcgacggc
      121 ctcatgagca caaaaaagaa accattaaca caagagcagc ttgaggacgc acgtcgcctt
      181 aaagcaattt atgaaaaaaa gaaaaatgaa cttggcttat cccaggaatc tgtcgcagac
      241 aagatgggga tggggcagtc aggcgttggt gctttattta atggcatcaa tgcattaaat
      301 gcttataacg ccgcattgct tgcaaaaatt ctcaaagtta gcgttgaaga atttagccct
      361 tcaatcgcca gagaaatcta cgagatgtat gaagcggtta gtatgcagcc gtcacttaga
      421 agtgagtatg agtaccctgt tttttctcat gttcaggcag ggatgttctc acctgagctt
      481 agaaccttta ccaaaggtga tgcggagaga tgggtaagca caaccaaaaa agccagtgat
      541 tctgcattct ggcttgaggt tgaaggtaat tccatgaccg caccaacagg ctccaagcca
      601 agctttcctg acggaatgtt aattctcgtt gaccctgagc aggctgttga gccaggtgat
      661 ttctgcatag ccagacttgg gggtgatgag tttaccttca agaaactgat cagggatagc
      721 ggtcaggtgt ttttacaacc actaaaccca cagtacccaa tgatcccatg caatgagagt
      781 tgttccgttg tggggaaagt tatcgctagt cagtggcctg aagagacgtt tggctaataa
      841 ggatctcaag ccgcagctaa aaggtaggat atacaagacc aggatctatg tccagattag
      901 ataaaagtaa agtgattaac agcgcattag agctgcttaa tgaggtcgga atcgaaggtt
      961 taacaacccg taaactcgcc cagaagctag gtgtagagca gcctacattg tattggcatg
     1021 taaaaaataa gcgggctttg ctcgacgcct tagccattga gatgttagat aggcaccata
     1081 ctcacttttg ccctttagaa ggggaaagct ggcaagattt tttacgtaat aacgctaaaa
     1141 gttttagatg tgctttacta agtcatcgcg atggagcaaa agtacattta ggtacacggc
     1201 ctacagaaaa acagtatgaa actctcgaaa atcaattagc ctttttatgc caacaaggtt
     1261 tttcactaga gaatgcatta tatgcactca gcgctgtggg gcattttact ttaggttgcg
     1321 tattggaaga tcaagagcat caagtcgcta aagaagaaag ggaaacacct actactgata
     1381 gtatgccgcc attattacga caagctatcg aattatttga tcaccaaggt gcagagccag
     1441 ccttcttatt cggccttgaa ttgatcatat gcggattaga aaaacaactt aaatgtgaaa
     1501 gtgggtccgc tgcaaacgac gaaaactacg ctttagtagc ttaataagga tcctaactcg
     1561 agtaaggatc tccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     1621 ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg
     1681 tgggcctttc tgcgtttata cctagggtac gggttttgct gcccgcaaac gggctgttct
     1741 ggtgttgcta gtttgttatc agaatcgcag atccggcttc agccggtttg ccggctgaaa
     1801 gcgctatttc ttccagaatt gccatgattt tttccccacg ggaggcgtca ctggctcccg
     1861 tgttgtcggc agctttgatt cgataagcag catcgcctgt ttcaggctgt ctatgtgtga
     1921 ctgttgagct gtaacaagtt gtctcaggtg ttcaatttca tgttctagtt gctttgtttt
     1981 actggtttca cctgttctat taggtgttac atgctgttca tctgttacat tgtcgatctg
     2041 ttcatggtga acagctttga atgcaccaaa aactcgtaaa agctctgatg tatctatctt
     2101 ttttacaccg ttttcatctg tgcatatgga cagttttccc tttgatatgt aacggtgaac
     2161 agttgttcta cttttgtttg ttagtcttga tgcttcactg atagatacaa gagccataag
     2221 aacctcagat ccttccgtat ttagccagta tgttctctag tgtggttcgt tgtttttgcg
     2281 tgagccatga gaacgaacca ttgagatcat acttactttg catgtcactc aaaaattttg
     2341 cctcaaaact ggtgagctga atttttgcag ttaaagcatc gtgtagtgtt tttcttagtc
     2401 cgttatgtag gtaggaatct gatgtaatgg ttgttggtat tttgtcacca ttcattttta
     2461 tctggttgtt ctcaagttcg gttacgagat ccatttgtct atctagttca acttggaaaa
     2521 tcaacgtatc agtcgggcgg cctcgcttat caaccaccaa tttcatattg ctgtaagtgt
     2581 ttaaatcttt acttattggt ttcaaaaccc attggttaag ccttttaaac tcatggtagt
     2641 tattttcaag cattaacatg aacttaaatt catcaaggct aatctctata tttgccttgt
     2701 gagttttctt ttgtgttagt tcttttaata accactcata aatcctcata gagtatttgt
     2761 tttcaaaaga cttaacatgt tccagattat attttatgaa tttttttaac tggaaaagat
     2821 aaggcaatat ctcttcacta aaaactaatt ctaatttttc gcttgagaac ttggcatagt
     2881 ttgtccactg gaaaatctca aagcctttaa ccaaaggatt cctgatttcc acagttctcg
     2941 tcatcagctc tctggttgct ttagctaata caccataagc attttcccta ctgatgttca
     3001 tcatctgagc gtattggtta taagtgaacg ataccgtccg ttctttcctt gtagggtttt
     3061 caatcgtggg gttgagtagt gccacacagc ataaaattag cttggtttca tgctccgtta
     3121 agtcatagcg actaatcgct agttcatttg ctttgaaaac aactaattca gacatacatc
     3181 tcaattggtc taggtgattt taatcactat accaattgag atgggctagt caatgataat
     3241 tactagtcct tttcccgggt gatctgggta tctgtaaatt ctgctagacc tttgctggaa
     3301 aacttgtaaa ttctgctaga ccctctgtaa attccgctag acctttgtgt gttttttttg
     3361 tttatattca agtggttata atttatagaa taaagaaaga ataaaaaaag ataaaaagaa
     3421 tagatcccag ccctgtgtat aactcactac tttagtcagt tccgcagtat tacaaaagga
     3481 tgtcgcaaac gctgtttgct cctctacaaa acagacctta aaaccctaaa ggcttaagta
     3541 gcaccctcgc aagctcgggc aaatcgctga atattccttt tgtctccgac catcaggcac
     3601 ctgagtcgct gtctttttcg tgacattcag ttcgctgcgc tcacggctct ggcagtgaat
     3661 gggggtaaat ggcactacag gcgcctttta tggattcatg caaggaaact acccataata
     3721 caagaaaagc ccgtcacggg cttctcaggg cgttttatgg cgggtctgct atgtggtgct
     3781 atctgacttt ttgctgttca gcagttcctg ccctctgatt ttccagtctg accacttcgg
     3841 attatcccgt gacaggtcat tcagactggc taatgcaccc agtaaggcag cggtatcatc
     3901 aacaggctta cccgtcttac tgtccctagt gcttggattc tcaccaataa aaaacgcccg
     3961 gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat
     4021 caacaggagt ccaagcgagc tctcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga
     4081 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
     4141 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
     4201 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
     4261 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg
     4321 ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
     4381 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
     4441 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
     4501 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
     4561 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
     4621 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt
     4681 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     4741 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     4801 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga
     4861 aacgatcctc atcctgtctc ttgatcagat catgatcccc tgcgccatca gatccttggc
     4921 ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca
     4981 gctggcaatt ccgaattcat gagatct
//
