LOCUS       40924_605        4234 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector DH6-1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4234)
  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 4234)
  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 4234)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4234)
  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..4234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(15..36)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(52..1131)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(1132..1209)
                     /label=lacI promoter
     protein_bind    1502..1518
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    1525..1541
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     5'UTR           1569..1603
     CDS             1604..2314
                     /label=lambda repressor
                     /note="phage lambda repressor"
     CDS             2315..2347
                     /label=ssrA tag (LVA)
                     /note="C-terminal peptide that mediates degradation in
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
     terminator      2386..2457
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2473..2500
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2658..3246)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3334..3428)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3452..4108)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(4109..4211)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
ORIGIN
        1 gacgtcggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
       61 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      121 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
      181 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
      241 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
      301 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt
      361 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
      421 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
      481 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
      541 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
      601 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
      661 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
      721 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
      781 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
      841 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
      901 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
      961 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     1021 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     1081 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     1141 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     1201 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     1261 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     1321 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     1381 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     1441 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     1501 attgtgagcg gataacaatt gacattgtga gcggataaca agatactgag cacgaattca
     1561 aaagatctaa tccaacaata tagcatactc tcgcgacggc ctcatgagca caaaaaagaa
     1621 accattaaca caagagcagc ttgaggacgc acgtcgcctt aaagcaattt atgaaaaaaa
     1681 gaaaaatgaa cttggcttat cccaggaatc tgtcgcagac aagatgggga tggggcagtc
     1741 aggcgttggt gctttattta atggcatcaa tgcattaaat gcttataacg ccgcattgct
     1801 tgcaaaaatt ctcaaagtta gcgttgaaga atttagccct tcaatcgcca gagaaatcta
     1861 cgagatgtat gaagcggtta gtatgcagcc gtcacttaga agtgagtatg agtaccctgt
     1921 tttttctcat gttcaggcag ggatgttctc acctgagctt agaaccttta ccaaaggtga
     1981 tgcggagaga tgggtaagca caaccaaaaa agccagtgat tctgcattct ggcttgaggt
     2041 tgaaggtaat tccatgaccg caccaacagg ctccaagcca agctttcctg acggaatgtt
     2101 aattctcgtt gaccctgagc aggctgttga gccaggtgat ttctgcatag ccagacttgg
     2161 gggtgatgag tttaccttca agaaactgat cagggatagc ggtcaggtgt ttttacaacc
     2221 actaaaccca cagtacccaa tgatcccatg caatgagagt tgttccgttg tggggaaagt
     2281 tatcgctagt cagtggcctg aagagacgtt tggcgctgca aacgacgaaa actacgcttt
     2341 agtagcttaa taaggatcca aactcgagta aggatctcca ggcatcaaat aaaacgaaag
     2401 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac
     2461 tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttataccta gggcgttcgg
     2521 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2581 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2641 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2701 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2761 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2821 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2881 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2941 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3001 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3061 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     3121 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3181 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3241 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3301 cgttaaggga ttttggtcat gactagtgct tggattctca ccaataaaaa acgcccggcg
     3361 gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa
     3421 caggagtcca agcgagctcg atatcaaatt acgccccgcc ctgccactca tcgcagtact
     3481 gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct
     3541 gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa
     3601 cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc
     3661 agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt
     3721 tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt
     3781 ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag
     3841 ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacga aattccggat
     3901 gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt
     3961 tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt
     4021 gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg
     4081 tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata
     4141 actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta
     4201 cgtgccgatc aacgtctcat tttcgccaga tatc
//
