LOCUS       40924_600        5088 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector DH4-27, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5088)
  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 5088)
  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 5088)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5088)
  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..5088
                     /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"
     misc_feature    835..915
                     /gene="cI with mf degradation tag"
                     /label=mf-ssrA tag (LVA)
                     /note="mf-ssrA tag (LVA)"
     5'UTR           928..968
     CDS             969..1589
                     /label=TetR
                     /note="tetracycline repressor TetR"
     CDS             1590..1622
                     /label=ssrA tag (LVA)
                     /note="C-terminal peptide that mediates degradation in
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
     terminator      1661..1732
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1748..1775
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2309..3256)
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
     rep_origin      complement(3304..3526)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4018..4112)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4146..4937)
                     /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 tggcgcggcg
      841 aataaaaatg aagaaaatac caatgaagtg ccgaccttta tgctgaatgc gggccaggcg
      901 aattatgcgt ttgcgtaata aggatctcaa gccgcagcta aaaggtagga tatacaagac
      961 caggatctat gtccagatta gataaaagta aagtgattaa cagcgcatta gagctgctta
     1021 atgaggtcgg aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc
     1081 agcctacatt gtattggcat gtaaaaaata agcgggcttt gctcgacgcc ttagccattg
     1141 agatgttaga taggcaccat actcactttt gccctttaga aggggaaagc tggcaagatt
     1201 ttttacgtaa taacgctaaa agttttagat gtgctttact aagtcatcgc gatggagcaa
     1261 aagtacattt aggtacacgg cctacagaaa aacagtatga aactctcgaa aatcaattag
     1321 cctttttatg ccaacaaggt ttttcactag agaatgcatt atatgcactc agcgctgtgg
     1381 ggcattttac tttaggttgc gtattggaag atcaagagca tcaagtcgct aaagaagaaa
     1441 gggaaacacc tactactgat agtatgccgc cattattacg acaagctatc gaattatttg
     1501 atcaccaagg tgcagagcca gccttcttat tcggccttga attgatcata tgcggattag
     1561 aaaaacaact taaatgtgaa agtgggtccg ctgcaaacga cgaaaactac gctttagtag
     1621 cttaataagg atcctaactc gagtaaggat ctccaggcat caaataaaac gaaaggctca
     1681 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag
     1741 tcacactggc tcaccttcgg gtgggccttt ctgcgtttat acctagggta cgggttttgc
     1801 tgcccgcaaa cgggctgttc tggtgttgct agtttgttat cagaatcgca gatccggctt
     1861 cagccggttt gccggctgaa agcgctattt cttccagaat tgccatgatt ttttccccac
     1921 gggaggcgtc actggctccc gtgttgtcgg cagctttgat tcgataagca gcatcgcctg
     1981 tttcaggctg tctatgtgtg actgttgagc tgtaacaagt tgtctcaggt gttcaatttc
     2041 atgttctagt tgctttgttt tactggtttc acctgttcta ttaggtgtta catgctgttc
     2101 atctgttaca ttgtcgatct gttcatggtg aacagctttg aatgcaccaa aaactcgtaa
     2161 aagctctgat gtatctatct tttttacacc gttttcatct gtgcatatgg acagttttcc
     2221 ctttgatatg taacggtgaa cagttgttct acttttgttt gttagtcttg atgcttcact
     2281 gatagataca agagccataa gaacctcaga tccttccgta tttagccagt atgttctcta
     2341 gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc attgagatca tacttacttt
     2401 gcatgtcact caaaaatttt gcctcaaaac tggtgagctg aatttttgca gttaaagcat
     2461 cgtgtagtgt ttttcttagt ccgttatgta ggtaggaatc tgatgtaatg gttgttggta
     2521 ttttgtcacc attcattttt atctggttgt tctcaagttc ggttacgaga tccatttgtc
     2581 tatctagttc aacttggaaa atcaacgtat cagtcgggcg gcctcgctta tcaaccacca
     2641 atttcatatt gctgtaagtg tttaaatctt tacttattgg tttcaaaacc cattggttaa
     2701 gccttttaaa ctcatggtag ttattttcaa gcattaacat gaacttaaat tcatcaaggc
     2761 taatctctat atttgccttg tgagttttct tttgtgttag ttcttttaat aaccactcat
     2821 aaatcctcat agagtatttg ttttcaaaag acttaacatg ttccagatta tattttatga
     2881 atttttttaa ctggaaaaga taaggcaata tctcttcact aaaaactaat tctaattttt
     2941 cgcttgagaa cttggcatag tttgtccact ggaaaatctc aaagccttta accaaaggat
     3001 tcctgatttc cacagttctc gtcatcagct ctctggttgc tttagctaat acaccataag
     3061 cattttccct actgatgttc atcatctgag cgtattggtt ataagtgaac gataccgtcc
     3121 gttctttcct tgtagggttt tcaatcgtgg ggttgagtag tgccacacag cataaaatta
     3181 gcttggtttc atgctccgtt aagtcatagc gactaatcgc tagttcattt gctttgaaaa
     3241 caactaattc agacatacat ctcaattggt ctaggtgatt ttaatcacta taccaattga
     3301 gatgggctag tcaatgataa ttactagtcc ttttcccggg tgatctgggt atctgtaaat
     3361 tctgctagac ctttgctgga aaacttgtaa attctgctag accctctgta aattccgcta
     3421 gacctttgtg tgtttttttt gtttatattc aagtggttat aatttataga ataaagaaag
     3481 aataaaaaaa gataaaaaga atagatccca gccctgtgta taactcacta ctttagtcag
     3541 ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt
     3601 aaaaccctaa aggcttaagt agcaccctcg caagctcggg caaatcgctg aatattcctt
     3661 ttgtctccga ccatcaggca cctgagtcgc tgtctttttc gtgacattca gttcgctgcg
     3721 ctcacggctc tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt atggattcat
     3781 gcaaggaaac tacccataat acaagaaaag cccgtcacgg gcttctcagg gcgttttatg
     3841 gcgggtctgc tatgtggtgc tatctgactt tttgctgttc agcagttcct gccctctgat
     3901 tttccagtct gaccacttcg gattatcccg tgacaggtca ttcagactgg ctaatgcacc
     3961 cagtaaggca gcggtatcat caacaggctt acccgtctta ctgtccctag tgcttggatt
     4021 ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc
     4081 tgaggtcatt actggatcta tcaacaggag tccaagcgag ctctcgaacc ccagagtccc
     4141 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
     4201 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
     4261 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg
     4321 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
     4381 acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc
     4441 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     4501 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     4561 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     4621 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     4681 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     4741 cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     4801 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     4861 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     4921 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcaga tcatgatccc
     4981 ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca
     5041 accttaccag agggcgcccc agctggcaat tccgaattca tgagatct
//
