LOCUS       40924_16790        5887 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pECJ3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5887)
  AUTHORS   Cameron DE, Collins JJ.
  TITLE     Tunable protein degradation in bacteria
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5887)
  AUTHORS   Cameron DE, Collins JJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-SEP-2014) Biomedical Engineering, Boston University, 
            44 Cummington St., Boston, MA 02215, USA
REFERENCE   3  (bases 1 to 5887)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5887)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-SEP-2014) Biomedical Engineering, Boston University, 44 
            Cummington St., Boston, MA 02215, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    49..54
                     /label=PvuII
                     /note="PvuII"
     CDS             complement(58..771)
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
     misc_feature    772..777
                     /label=ApaLI
                     /note="ApaLI"
     misc_feature    817..822
                     /label=SphI
                     /note="SphI"
     regulatory      complement(823..896)
                     /label=PLtetO promoter
                     /note="PLtetO promoter"
                     /regulatory_class="promoter"
     promoter        complement(823..896)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
     misc_feature    842
                     /label=transcription start
                     /note="transcription start"
     protein_bind    853..871
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    878..896
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    897..902
                     /label=AclI
                     /note="AclI"
     misc_feature    933..938
                     /label=SacII
                     /note="SacII"
     misc_feature    complement(939..1022)
                     /label=pdt#3
                     /note="pdt#3"
     CDS             complement(1023..2102)
                     /label=lacI
                     /note="lac repressor"
     misc_feature    2112..2117
                     /label=BsrGI
                     /note="BsrGI"
     misc_feature    2142..2147
                     /label=BamHI
                     /note="BamHI"
     promoter        complement(2148..2221)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
     misc_feature    2222..2227
                     /label=EagI
                     /note="EagI"
     misc_feature    2234..2239
                     /label=NcoI
                     /note="NcoI"
     promoter        2253..2282
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     misc_feature    2288
                     /label=transcription start
                     /note="transcription start"
     protein_bind    2290..2306
                     /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)."
     misc_feature    2315..2320
                     /label=SalI
                     /note="SalI"
     regulatory      2337..2340
                     /label=RBS8
                     /note="RBS8"
                     /regulatory_class="ribosome_binding_site"
     misc_feature    2347..2352
                     /label=NheI
                     /note="NheI"
     CDS             2353..2973
                     /label=TetR
                     /note="tetracycline repressor TetR"
     misc_feature    2977..2982
                     /label=SacI
                     /note="SacI"
     misc_feature    3001..3006
                     /label=NdeI
                     /note="NdeI"
     misc_feature    3013..3018
                     /label=XmaI
                     /note="XmaI"
     regulatory      3019..3093
                     /label=Ptrc-2 promoter
                     /note="Ptrc-2 promoter"
                     /regulatory_class="promoter"
     promoter        3032..3061
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    3069..3085
                     /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)."
     misc_feature    3069
                     /label=transcription start
                     /note="transcription start"
     misc_feature    3094..3099
                     /label=MfeI
                     /note="MfeI"
     misc_feature    3129..3134
                     /label=SfoI
                     /note="SfoI"
     CDS             3136..3843
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     misc_feature    3847..3852
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    3859..3864
                     /label=AvrII
                     /note="AvrII"
     misc_feature    3871..3876
                     /label=XhoI
                     /note="XhoI"
     terminator      3906..3992
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     rep_origin      complement(4156..4744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4832..4926)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4960..5751)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 gacgtctgtg caagtactac tgttctgcag tcacttgaat tcgataccca gctgttattt
       61 gtatagttca tccatgccat gtgtaatccc agcagcggtt acaaactcaa gaaggaccat
      121 gtggtctctc ttttcgttgg gatctttcga aagggcagat tgtgtggaca ggtaatggtt
      181 gtctggtaaa aggacagggc catcgccaat cggagtattt tgttgataat ggtctgctag
      241 ttgaacgctt ccatcttcaa tgttgtgtct aattttgaag ttaactttga ttccattctt
      301 ttgtttgtct gccatgatgt atacattgtg tgagttatag ttgtattcca atttgtgtcc
      361 aagaatgttt ccatcttctt taaaatcaat accttttaac tcgattctat taacaagggt
      421 atcaccttca aacttgactt cagcacgtgt cttgtagttc ccgtcatctt tgaaaaatat
      481 agttctttcc tgtacgtaac cttcgggcat ggcactcttg aaaaagtcat gctgtttcat
      541 gtgatctggg tatctcgcaa agcattgaac accataaccg aaagtagtga caagtgttgg
      601 ccacggaaca ggtagttttc cagtagtgca aataaattta agggtaagtt ttccgtatgt
      661 tgcatcacct tcaccctctc cactgacaga aaatttgtgc ccattaacat caccatctaa
      721 ttcaacaaga attgggacaa ctccagtgaa aagttcttct cctttacgca tgtgcactac
      781 ctccttggtt gtctatgcta tgctgatcta caactggcat gcggtcagtg cgtcctgctg
      841 atgtgctcag tatctctatc actgataggg atgtcaatct ctatcactga tagggaaacg
      901 tttcgcagaa gcttccgcaa ggtaccactt tgccgcggtt aaactcgtct tctgttcgcc
      961 tggcccgcgt tcagcataaa ggtcggcact tcgttggtgt tttcttcgtt tttgttcgcc
     1021 gcctgcccgc tttccagtcg ggaaacctgt cgtgccagtt gcattaatga atcggccaac
     1081 gcgcggggag aggcggtttg cgtattggga gccagggtgg tttttctttt caccagtgag
     1141 acgggcaaca gttgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     1201 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     1261 catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc
     1321 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     1381 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     1441 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     1501 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     1561 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     1621 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     1681 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     1741 ccactgacgc gctgcgcgag aagattgtga accgctgctt tacaggcttc gacgccgctt
     1801 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     1861 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     1921 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     1981 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     2041 gaaacggtct gataagagac accggcatac tctgcgacat cgtacagcgt tactggtttc
     2101 acattcacca ttgtacaccc tctccttata ttgggaccta tggatccggt cagtgcgtcc
     2161 tgctgatgtg ctcagtatct ctatcactga tagggatgtc aatctctatc actgataggg
     2221 acggccgccc gttccatggc tgaaatgagc tgttgacaat taatcatccg gctcgtataa
     2281 tgtgtggaat tgtgagcgga taacaatttc acacgtcgac cgccttcggc gaagctccac
     2341 acgagagcta gcatgtctcg tttagataaa agtaaagtga ttaacagcgc attagagctg
     2401 cttaatgagg tcggaatcga aggtttaaca acccgtaaac tcgcccagaa gctaggtgta
     2461 gagcagccta cattgtattg gcatgtaaaa aataagcggg ctttgctcga cgccttagcc
     2521 attgagatgt tagataggca ccatactcac ttttgccctt tagaagggga aagctggcaa
     2581 gattttttac gtaataacgc taaaagtttt agatgtgctt tactaagtca tcgcgatgga
     2641 gcaaaagtac atttaggtac acggcctaca gaaaaacagt atgaaactct cgaaaatcaa
     2701 ttagcctttt tatgccaaca aggtttttca ctagagaatg cattatatgc actcagcgct
     2761 gtggggcatt ttactttagg ttgcgtattg gaagatcaag agcatcaagt cgctaaagaa
     2821 gaaagggaaa cacctactac tgatagtatg ccgccattat tacgacaagc tatcgaatta
     2881 tttgatcacc aaggtgcaga gccagccttc ttattcggcc ttgaattgat catctgcgga
     2941 ttagaaaaac aacttaaatg tgaaagtggg tcttgagagc tcggactgct taagtcgctc
     3001 catatgctcg ttcccgggct gaaatgagct gttgacaatt aatcatccgg ctcgtataat
     3061 gtgtggaatt gtgagcggat aacaatttca caccaattga gtattaacta tcgttcaact
     3121 gatagggagg cgccgatggt gagcaagggc gaggaggata acatggccat catcaaggag
     3181 ttcatgcgct tcaaggttca catggagggc tccgtgaacg gccacgagtt cgagatcgag
     3241 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     3301 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
     3361 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc
     3421 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
     3481 tcctccctgc aagacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc
     3541 tccgacggcc ccgtaatgca gaagaagact atgggctggg aggcctcctc cgagcggatg
     3601 taccccgagg acggcgcgct gaagggcgag atcaagcaga ggctgaagct gaaggacggc
     3661 ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt gcaactgccc
     3721 ggcgcgtaca acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc
     3781 gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtat
     3841 aagtaagata tctatcgccc tagggaccgt ctcgagagaa tcaatattaa tccaacgcgt
     3901 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     3961 tgtcggtgaa cgctctcctg agtaggacaa atccgccgcc ctagacttag gcgttcggct
     4021 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4081 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4141 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4201 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4261 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4321 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4381 gtaggtcgtt cgctccaagc tgggctgtct gcacgaaccc cccgttcagc ccgaccgctg
     4441 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4501 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4561 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     4621 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4681 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4741 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4801 ttaagggatt ttggtcatgg ctagtgcttg gattctcacc aataaaaaac gcccggcggc
     4861 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca
     4921 ggagtccaag ccaattctcg aaccccagag tcccgctcag aagaactcgt caagaaggcg
     4981 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     5041 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     5101 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     5161 catgatattc ggcaagcagg catcgccgtg ggtcacgacg agatcctcgc cgtcgggcat
     5221 acgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     5281 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     5341 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     5401 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     5461 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagccgc
     5521 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt
     5581 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgaccctgcg ctgacagccg
     5641 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     5701 ctccacccaa gccgccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga
     5761 tcctcatcct gtctcttgat cagatcttga tcccctgcgc catcagatcc ttggcggcaa
     5821 gaaagccatc cagtttactt tgcagggctt cccaacctta ccagagggcg gcccaactgg
     5881 caattcc
//
