LOCUS       40924_625        5040 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector DH6-64, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5040)
  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 5040)
  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 5040)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5040)
  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..5040
                     /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"
     CDS             835..867
                     /label=ssrA tag (LVA)
                     /note="C-terminal peptide that mediates degradation in
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
     5'UTR           880..920
     CDS             921..1541
                     /label=TetR
                     /note="tetracycline repressor TetR"
     CDS             1542..1574
                     /label=ssrA tag (LVA)
                     /note="C-terminal peptide that mediates degradation in
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
     terminator      1613..1684
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1700..1727
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2261..3208)
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
     rep_origin      complement(3256..3478)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3970..4064)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4098..4889)
                     /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 tggcgctgca
      841 aacgacgaaa actacgcttt agtagcttaa taaggatctc aagccgcagc taaaaggtag
      901 gatatacaag accaggatct atgtccagat tagataaaag taaagtgatt aacagcgcat
      961 tagagctgct taatgaggtc ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc
     1021 taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg
     1081 ccttagccat tgagatgtta gataggcacc atactcactt ttgcccttta gaaggggaaa
     1141 gctggcaaga ttttttacgt aataacgcta aaagttttag atgtgcttta ctaagtcatc
     1201 gcgatggagc aaaagtacat ttaggtacac ggcctacaga aaaacagtat gaaactctcg
     1261 aaaatcaatt agccttttta tgccaacaag gtttttcact agagaatgca ttatatgcac
     1321 tcagcgctgt ggggcatttt actttaggtt gcgtattgga agatcaagag catcaagtcg
     1381 ctaaagaaga aagggaaaca cctactactg atagtatgcc gccattatta cgacaagcta
     1441 tcgaattatt tgatcaccaa ggtgcagagc cagccttctt attcggcctt gaattgatca
     1501 tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc cgctgcaaac gacgaaaact
     1561 acgctttagt agcttaataa ggatcctaac tcgagtaagg atctccaggc atcaaataaa
     1621 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     1681 tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt atacctaggg
     1741 tacgggtttt gctgcccgca aacgggctgt tctggtgttg ctagtttgtt atcagaatcg
     1801 cagatccggc ttcagccggt ttgccggctg aaagcgctat ttcttccaga attgccatga
     1861 ttttttcccc acgggaggcg tcactggctc ccgtgttgtc ggcagctttg attcgataag
     1921 cagcatcgcc tgtttcaggc tgtctatgtg tgactgttga gctgtaacaa gttgtctcag
     1981 gtgttcaatt tcatgttcta gttgctttgt tttactggtt tcacctgttc tattaggtgt
     2041 tacatgctgt tcatctgtta cattgtcgat ctgttcatgg tgaacagctt tgaatgcacc
     2101 aaaaactcgt aaaagctctg atgtatctat cttttttaca ccgttttcat ctgtgcatat
     2161 ggacagtttt ccctttgata tgtaacggtg aacagttgtt ctacttttgt ttgttagtct
     2221 tgatgcttca ctgatagata caagagccat aagaacctca gatccttccg tatttagcca
     2281 gtatgttctc tagtgtggtt cgttgttttt gcgtgagcca tgagaacgaa ccattgagat
     2341 catacttact ttgcatgtca ctcaaaaatt ttgcctcaaa actggtgagc tgaatttttg
     2401 cagttaaagc atcgtgtagt gtttttctta gtccgttatg taggtaggaa tctgatgtaa
     2461 tggttgttgg tattttgtca ccattcattt ttatctggtt gttctcaagt tcggttacga
     2521 gatccatttg tctatctagt tcaacttgga aaatcaacgt atcagtcggg cggcctcgct
     2581 tatcaaccac caatttcata ttgctgtaag tgtttaaatc tttacttatt ggtttcaaaa
     2641 cccattggtt aagcctttta aactcatggt agttattttc aagcattaac atgaacttaa
     2701 attcatcaag gctaatctct atatttgcct tgtgagtttt cttttgtgtt agttctttta
     2761 ataaccactc ataaatcctc atagagtatt tgttttcaaa agacttaaca tgttccagat
     2821 tatattttat gaattttttt aactggaaaa gataaggcaa tatctcttca ctaaaaacta
     2881 attctaattt ttcgcttgag aacttggcat agtttgtcca ctggaaaatc tcaaagcctt
     2941 taaccaaagg attcctgatt tccacagttc tcgtcatcag ctctctggtt gctttagcta
     3001 atacaccata agcattttcc ctactgatgt tcatcatctg agcgtattgg ttataagtga
     3061 acgataccgt ccgttctttc cttgtagggt tttcaatcgt ggggttgagt agtgccacac
     3121 agcataaaat tagcttggtt tcatgctccg ttaagtcata gcgactaatc gctagttcat
     3181 ttgctttgaa aacaactaat tcagacatac atctcaattg gtctaggtga ttttaatcac
     3241 tataccaatt gagatgggct agtcaatgat aattactagt ccttttcccg ggtgatctgg
     3301 gtatctgtaa attctgctag acctttgctg gaaaacttgt aaattctgct agaccctctg
     3361 taaattccgc tagacctttg tgtgtttttt ttgtttatat tcaagtggtt ataatttata
     3421 gaataaagaa agaataaaaa aagataaaaa gaatagatcc cagccctgtg tataactcac
     3481 tactttagtc agttccgcag tattacaaaa ggatgtcgca aacgctgttt gctcctctac
     3541 aaaacagacc ttaaaaccct aaaggcttaa gtagcaccct cgcaagctcg ggcaaatcgc
     3601 tgaatattcc ttttgtctcc gaccatcagg cacctgagtc gctgtctttt tcgtgacatt
     3661 cagttcgctg cgctcacggc tctggcagtg aatgggggta aatggcacta caggcgcctt
     3721 ttatggattc atgcaaggaa actacccata atacaagaaa agcccgtcac gggcttctca
     3781 gggcgtttta tggcgggtct gctatgtggt gctatctgac tttttgctgt tcagcagttc
     3841 ctgccctctg attttccagt ctgaccactt cggattatcc cgtgacaggt cattcagact
     3901 ggctaatgca cccagtaagg cagcggtatc atcaacaggc ttacccgtct tactgtccct
     3961 agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc
     4021 cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg agctctcgaa
     4081 ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa
     4141 tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct
     4201 tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg
     4261 ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca
     4321 tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag cctggcgaac
     4381 agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg
     4441 gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag
     4501 gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg
     4561 gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag
     4621 tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc
     4681 agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga caggtcggtc
     4741 ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag
     4801 ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa
     4861 cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatca
     4921 gatcatgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca gtttactttg
     4981 cagggcttcc caaccttacc agagggcgcc ccagctggca attccgaatt catgagatct
//
