LOCUS       40924_16795       10199 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pECL275, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10199)
  AUTHORS   Cameron DE, Collins JJ.
  TITLE     Tunable protein degradation in bacteria
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10199)
  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 10199)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10199)
  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            "Unpublished"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..10199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             7..177
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
     terminator      455..482
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        604..677
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
     RBS             688..696
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             700..3063
                     /codon_start=1
                     /product="mf-lon"
                     /label=mf-lon
                     /protein_id="AIU94520.1"
                     /translation="MSKKIKLPIFQIRGSFIVPGIKENLEVGRKNTLASVNYAIKNSNN
                     QMIAIPQIDASVEKPEFSDLHEFGILIDFEVIKEWKDNSLTISTNPIQRCKVISFFENE
                     DQVPYAEVELIESINDFSDEELKELIEKISDAIKTKASLVTKQIKQLISGESDDLSLAF
                     DSIMFKLAPSKILTNPEYITSPSLKTRWSIIEKIIFAEDGIITRNAESIDAARQKNEIE
                     QELNHKLKEKMDKQQKEYYLREKMRIIKDELEDEDDSDDSSLEKYKERLAKEPFPEEVK
                     RKIMASIKRVEALQSGTPEWNTEKNYIDWMMSIPWWEETEDLTDLKYAKKILDKHHYGM
                     KKVKERIIEYLAVKTKTKSLKAPIITLVGPPGVGKTSLAKSIAEAVGKNFVKVSLGGVK
                     DESEIRGHRKTYVGSMPGRIIQTMKRAKVKNPLFLLDEIDKMASDHRGDPASAMLEVLD
                     PEQNKEFSDHYIEEPYDLSQVMFIATANYPEDIPEALYDRMEIINLSSYTEIEKVKIAQ
                     DYLVPKAIEQHELTSEEISFTEGAINEIIKYYTREAGVRQLERHINSIIRKYIVKNLNG
                     EMDKIVIDEKQVNDLLGKRIFDHTEKQEESQIGVVTGLAYTQFGGDILPIEVSLYPGKG
                     NLILTGKLGEVMKESATIALTYVKSNFEKFGVDKKVFEENDIHVHVPEGAVPKDGPSAG
                     ITITTALISALSDKPVSKEIGMTGEITLRGNVLPIGGLREKSISASRSGLKTIIIPKKN
                     ERDLDEIPDEVKAKLKIIPAEKYEEVFAIVFKTK"
     terminator      3106..3192
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    3208..3621
                     /note="lacZ; 3' region"
     rep_origin      3922..4310
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     promoter        4420..4524
                     /label=AmpR promoter
     CDS             4525..5382
                     /label=AmpR
                     /note="beta-lactamase"
     CDS             complement(6955..8373)
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     promoter        complement(8374..8819)
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     misc_feature    9851..10094
                     /note="oriT; RP4"
     oriT            9986..10095
                     /label=incP origin of transfer
                     /note="incP origin of transfer"
ORIGIN
        1 actagtacca tgattacgga ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
       61 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
      121 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
      181 cgctttgcct ggtttccggc accagaagcg gtgccggaaa gctggctgga gtgcgatctt
      241 cctgaggccg atactgtcgt cgtcccctca aactggcaga tgcacggtta cgatgcgccc
      301 atctacacca acgtgaccta tcccattacg gtcaatccgc cgtttgttcc cacggagaat
      361 ccgacgggtt gttactcgct cacatttaat gttgatgaaa gctggctaca ggaaggccag
      421 acgcggctag ccaaatccgc cgctctagct aagcagaagg ccatcctgac ggatggcctt
      481 tttgcgtttc tacaaactct tgttggggat ccgcctgcag cctacgtcta agaaaccatt
      541 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcacctc
      601 gagtccctat cagtgataga gattgacatc cctatcagtg atagagatac tgagcacatc
      661 agcaggacgc actgaccgaa ttcgcattaa ggaggtacca tgagtaaaaa aatcaaacta
      721 cctattttcc aaatccgtgg ttctttcatc gttccgggta tcaaagaaaa cctggaagtt
      781 ggtcgtaaaa acaccctggc ttctgttaac tacgctatca aaaactctaa caaccagatg
      841 atcgctatcc cgcagatcga cgcttctgtt gaaaaaccgg agttctctga cctgcacgag
      901 ttcggtatcc tgatcgactt cgaagttatc aaagaatgga aagacaactc tctgaccatc
      961 tctaccaacc cgatccagcg ttgcaaagtt atctctttct tcgaaaacga agaccaggtt
     1021 ccgtacgctg aagttgaact gatcgaatct atcaacgact tctctgacga agaactgaaa
     1081 gaactgatcg aaaaaatctc tgacgctatc aaaaccaagg cttctctggt taccaaacag
     1141 atcaaacaac tgatctctgg tgaatctgac gacctgtctc tggctttcga ctctatcatg
     1201 ttcaaactgg ctccgtctaa aatcctgacc aacccggaat acatcacctc tccgtctctg
     1261 aaaacccgtt ggtctatcat cgaaaaaatc atcttcgctg aagacggtat catcacccgt
     1321 aacgctgaat ctatcgacgc tgctcgtcag aaaaacgaaa tcgaacagga actgaaccac
     1381 aaactgaaag aaaaaatgga caaacagcag aaagaatact acctgcgtga aaaaatgcgt
     1441 atcatcaaag acgaactgga agacgaagac gactctgacg actcttctct ggaaaaatac
     1501 aaagaacgtc tggctaaaga accgttcccg gaagaagtta aacgtaaaat catggcttct
     1561 atcaaacgtg ttgaagctct gcaatctgga accccggaat ggaacaccga aaaaaactac
     1621 atcgactgga tgatgtctat cccgtggtgg gaagaaaccg aagacctgac cgacctgaaa
     1681 tacgctaaaa aaatcctgga caaacaccac tacggtatga aaaaagttaa agaacgtatc
     1741 atcgaatacc tggctgttaa aaccaaaacc aaatctctga aagctccgat catcaccctg
     1801 gttggtccgc cgggtgttgg taaaacctct ctggctaaat ctatcgctga agctgttggt
     1861 aaaaacttcg ttaaagtttc tctgggtggt gttaaagacg aatctgaaat ccgtggtcac
     1921 cgtaaaacct acgttggttc tatgccgggt cgtatcatcc agaccatgaa acgtgctaaa
     1981 gttaaaaacc cgctgttcct gctggacgaa atcgacaaaa tggcttctga ccaccgtggt
     2041 gacccggctt ctgctatgct ggaagttctg gacccggaac agaacaaaga gttctctgac
     2101 cactacatcg aagaaccgta cgacctgtct caggttatgt tcatcgctac cgctaactac
     2161 ccggaagaca tcccggaagc tctgtacgac cgtatggaaa tcatcaacct gtcttcttac
     2221 accgaaatcg aaaaagttaa aatcgctcag gactacctgg ttccgaaagc tatcgaacag
     2281 cacgaactga cctctgaaga aatctctttc accgaaggtg ctatcaacga aatcatcaaa
     2341 tactacaccc gtgaagctgg tgttcgtcaa ctggaacgtc acatcaactc tatcatccgt
     2401 aaatacatcg ttaaaaacct gaacggtgaa atggacaaaa tcgttatcga cgaaaaacag
     2461 gttaacgacc tgctgggtaa acgtatcttc gaccacaccg aaaaacagga agaatctcag
     2521 atcggtgttg ttaccggtct ggcttacacc cagttcggtg gtgacatcct gccgattgaa
     2581 gtttctctgt atccgggtaa aggtaacctg atcctgaccg gtaaactggg tgaagttatg
     2641 aaagaatctg ctaccatcgc tctgacctac gttaaatcta acttcgaaaa attcggtgtt
     2701 gacaaaaaag ttttcgaaga aaacgacatc cacgttcacg ttccggaagg tgctgttccg
     2761 aaagacggtc cgtctgctgg tatcaccatc accaccgctc tgatctctgc tctttctgac
     2821 aaaccggttt ctaaagaaat cggtatgacc ggtgaaatca ccctgcgtgg taacgttctg
     2881 ccgattggtg gtctgcgtga aaaatctatc tctgcttctc gttctggtct gaaaaccatc
     2941 atcatcccga aaaaaaacga acgtgacctg gacgaaatcc cggacgaagt taaagctaaa
     3001 ctgaaaatca tcccggctga aaaatacgaa gaggtatttg caatagtttt taaaacaaaa
     3061 taataagctt gatgggggat cccatggtac gcgtgctaga ggcatcaaat aaaacgaaag
     3121 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     3181 agtaggacaa atccgccgcg gcgcgccgcg gattggcctg aactgccagc tggcgcaggt
     3241 agcagagcgg gtaaactggc tcggattagg gccgcaagaa aactatcccg accgccttac
     3301 tgccgcctgt tttgaccgct gggatctgcc attgtcagac atgtataccc cgtacgtctt
     3361 cccgagcgaa aacggtctgc gctgcgggac gcgcgaattg aattatggcc cacaccagtg
     3421 gcgcggcgac ttccagttca acatcagccg ctacagtcaa cagcaactga tggaaaccag
     3481 ccatcgccat ctgctgcacg cggaagaagg cacatggctg aatatcgacg gtttccatat
     3541 ggggattggt ggcgacgact cctggagccc gtcagtatcg gcggaattcc agctgagcgc
     3601 cggtcgctac cattaccagt tgcggccgcc acgtagtggg ccatcgccct gatagacggt
     3661 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3721 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3781 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3841 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac cagatccggc
     3901 cacgatgcgt ccggcgtaga ggatctgaag atcagcagtt caacctgttg atagtacgta
     3961 ctaagctctc atgtttcacg tactaagctc tcatgtttaa cgtactaagc tctcatgttt
     4021 aacgaactaa accctcatgg ctaacgtact aagctctcat ggctaacgta ctaagctctc
     4081 atgtttcacg tactaagctc tcatgtttga acaataaaat taatataaat cagcaactta
     4141 aatagcctct aaggttttaa gttttataag aaaaaaaaga atatataagg cttttaaagc
     4201 ttttaaggtt taacggttgt ggacaacaag ccagggatgt aacgcactga gaagccctta
     4261 gagcctctca aagcaatttt gagtgacaca ggaacactta acggctgaca tgggaattct
     4321 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4381 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4441 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4501 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4561 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     4621 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     4681 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     4741 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     4801 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     4861 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     4921 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     4981 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5041 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     5101 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5161 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5221 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5281 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5341 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5401 tactcatata tactttagat tgatttaaaa cttcattttt aatttatggt gcactctcag
     5461 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     5521 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     5581 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     5641 aggttttcac cgtcatcacc gaaacgcgcg aggcagccaa ggagatggcg cccaacagtc
     5701 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     5761 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     5821 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatctcggga aaagcgttgg
     5881 tgaccaaagg tgccttttat catcacttta aaaataaaaa acaattactc agtgcctgtt
     5941 ataagcagca attaattatg attgatgcct acatcacaac aaaaactgat ttaacaaatg
     6001 gttggtctgc cttagaaagt atatttgaac attatcttga ttatattatt gataataata
     6061 aaaaccttat ccctatccaa gaagtgatgc ctatcattgg ttggaatgaa cttgaaaaaa
     6121 ttagccttga atacattact ggtaaggtaa acgccattgt cagcaaattg atccaagaga
     6181 accaacttaa agcttatgat agtgatgtgc ttaaaaactt actcaatggc tggtttatgc
     6241 agttcactta caccgcttct caacccggta cgcaccagaa aatcattgat atggccatga
     6301 atggcgttgg atgccgggca acagcccgca ttatgggcgt tggcctcaac acgattttac
     6361 gtcacttaaa aaactcaggc cgcagtcggt aacctcgcgc atacagccgg gcagtgacgt
     6421 catcgtctgc gcggaaatgg acgaacagtg gggctatgtc ggggctaaat cgcgccagcg
     6481 ctggctgttt tacgcgtatg acagtctccg gaagacggtt gttgcgcacg tattcggtga
     6541 acgcactatg gcgacgctgg ggcgtcttat gagcctgctg tcaccctttg acgtggtgat
     6601 atggatgacg gatggctggc cgctgtatga atcccgcctg aagggaaagc tgcacgtaat
     6661 cagcaagcga tatacgcagc gaattgagcg gcataacctg aatctgaggc agcacctggc
     6721 acggctggga cggaagtcgc tgtcgttctc aaaatcggtg gagctgcatg acaaagtcat
     6781 cgggcattat ctgaacataa aacactatca ataagttgga gtcattacca aaaggttagg
     6841 aatacggtta gccatttgcc tgcttttata tagttcatat gggattcacc tttatgttga
     6901 taagaaataa aagaaaatgc caataggata tcggcatttt cttttgcgtt tttatttgtt
     6961 aactgttaat tgtccttgtt caaggatgct gtctttgaca acagatgttt tcttgccttt
     7021 gatgttcagc aggaagcttg gcgcaaacgt tgattgtttg tctgcgtaga atcctctgtt
     7081 tgtcatatag cttgtaatca cgacattgtt tcctttcgct tgaggtacag cgaagtgtga
     7141 gtaagtaaag gttacatcgt taggatcaag atccattttt aacacaaggc cagttttgtt
     7201 cagcggcttg tatgggccag ttaaagaatt agaaacataa ccaagcatgt aaatatcgtt
     7261 agacgtaatg ccgtcaatcg tcatttttga tccgcgggag tcagtgaaca ggtaccattt
     7321 gccgttcatt ttaaagacgt tcgcgcgttc aatttcatct gttactgtgt tagatgcaat
     7381 cagcggtttc atcacttttt tcagtgtgta atcatcgttt agctcaatca taccgagagc
     7441 gccgtttgct aactcagccg tgcgtttttt atcgctttgc agaagttttt gactttcttg
     7501 acggaagaat gatgtgcttt tgccatagta tgctttgtta aataaagatt cttcgccttg
     7561 gtagccatct tcagttccag tgtttgcttc aaatactaag tatttgtggc ctttatcttc
     7621 tacgtagtga ggatctctca gcgtatggtt gtcgcctgag ctgtagttgc cttcatcgat
     7681 gaactgctgt acattttgat acgtttttcc gtcaccgtca aagattgatt tataatcctc
     7741 tacaccgttg atgttcaaag agctgtctga tgctgatacg ttaacttgtg cagttgtcag
     7801 tgtttgtttg ccgtaatgtt taccggagaa atcagtgtag aataaacgga tttttccgtc
     7861 agatgtaaat gtggctgaac ctgaccattc ttgtgtttgg tcttttagga tagaatcatt
     7921 tgcatcgaat ttgtcgctgt ctttaaagac gcggccagcg tttttccagc tgtcaataga
     7981 agtttcgccg actttttgat agaacatgta aatcgatgtg tcatccgcat ttttaggatc
     8041 tccggctaat gcaaagacga tgtggtagcc gtgatagttt gcgacagtgc cgtcagcgtt
     8101 ttgtaatggc cagctgtccc aaacgtccag gccttttgca gaagagatat ttttaattgt
     8161 ggacgaatcg aattcaggaa cttgatattt ttcatttttt tgctgttcag ggatttgcag
     8221 catatcatgg cgtgtaatat gggaaatgcc gtatgtttcc ttatatggct tttggttcgt
     8281 ttctttcgca aacgcttgag ttgcgcctcc tgccagcagt gcggtagtaa aggttaatac
     8341 tgttgcttgt tttgcaaact ttttgatgtt catcgttcat gtctcctttt ttatgtactg
     8401 tgttagcggt ctgcttcttc cagccctcct gtttgaagat ggcaagttag ttacgcacaa
     8461 taaaaaaaga cctaaaatat gtaaggggtg acgccaaagt atacactttg ccctttacac
     8521 attttaggtc ttgcctgctt tatcagtaac aaacccgcgc gatttacttt tcgacctcat
     8581 tctattagac tctcgtttgg attgcaactg gtctattttc ctcttttgtt tgatagaaaa
     8641 tcataaaagg atttgcagac tacgggccta aagaactaaa aaatctatct gtttcttttc
     8701 attctctgta ttttttatag tttctgttgc atgggcataa agttgccttt ttaatcacaa
     8761 ttcagaaaat atcataatat ctcatttcac taaataatag tgaacggcag gtatatgtga
     8821 tgggttaaaa aggatcgatc ctctagagtc gacctgcatt atatgcactc agcgctgtgg
     8881 ggcattttac tttaggttgc gtattggaag atcaagagca tcaagtcgct aaagaagaaa
     8941 gggaaacacc tactactgat agtatgccgc cattattacg acaagctatc gaattatttg
     9001 atcaccaagg tgcagagcca gccttcttat tcggccttga attgatcata tgcggattag
     9061 aaaaacaact taaatgtgaa agtgggtctt aaaagcagca taaccttttt ccgtgatggt
     9121 aacttcacgg taaccaagat gtcgagttaa ccacccttta gattcataaa gcgaaaataa
     9181 tgcggctcca acgtacccac ctaaatggaa acggcgttca ctccaatcta aacacgcaca
     9241 acagatttta cgtgaatgtt tggaaggaac gtcaattccc atttcatgaa aatattgaat
     9301 accacttaat gtgatcattg aaccattttc agtgatccat tgctgttgac aaagggaatc
     9361 atagatcctt tttgtccggt gttgggttga aggtgaagcc ggtcggggcc gcagcggggg
     9421 ccggcttttc agccttgccc ccctgcttcg gccgccgtgg ctccggcgtc ttgggtgccg
     9481 gcgcgggttc cgcagccttg gcctgcggtg cgggcacatc ggcgggcttg gccttgatgt
     9541 gccgcctggc gtgcgagcgg aacgtctcgt aggagaactt gaccttcccc gtttcccgca
     9601 tgtgctccca aatggtgacg agcgcatagc cggacgctaa cgccgcctcg acatccgccc
     9661 tcaccgccag gaacgcaacc gcagcctcat cacgccggcg cttcttggcc gcgcgggatt
     9721 caacccactc ggccagctcg tcggtgtagc tctttggcat cgtctctcgc ctgtcccctc
     9781 agttcagtaa tttcctgcat ttgcctgttt ccagtcggta gatattccac aaaacagcag
     9841 ggaagcagcg cttttccgct gcataaccct gcttcggggt cattatagcg attttttcgg
     9901 tatatccatc ctttttcgca cgatatacag gattttgcca aagggttcgt gtagactttc
     9961 cttggtgtat ccaacggcgt cagccgggca ggataggtga agtaggccca cccgcgagcg
    10021 ggtgttcctt cttcactgtc ccttattcgc acctggcggt gctcaacggg aatcctgctc
    10081 tgcgaggctg gccggctacc gccggcgtaa cagatgaggg caagcggatg gctgatgaaa
    10141 ccaagccaac caggaagggc agcccaccta tcaaggtgta ctgccttcca gacgaagtg
//
