LOCUS       40924_16700        7724 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pECA102, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7724)
  AUTHORS   Cameron DE, Collins JJ.
  TITLE     Tunable protein degradation in bacteria
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7724)
  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 7724)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7724)
  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            "Unpublished"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..7724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(100..975)
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1002..1286
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     CDS             1343..2611
                     /codon_start=1
                     /label=FLPo
                     /note="nuclear-targeted site-specific recombinase"
                     /translation="MSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAAELTYLCWMI
                     THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWGFTI
                     IPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
                     SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
                     KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
                     SYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
                     TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
                     YPAWNGIISQEVLDYLSSYINRRI"
     promoter        2670..3115
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             3116..4534
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     terminator      4752..4838
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4930..4957
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        4976..5067
                     /label=AmpR promoter
     CDS             5068..5925
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5970..6425
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      6536..7124
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 catcgatgca taatgtgcct gtcaaatgga cgaagcaggg attctgcaaa ccctatgcta
       61 ctccgtcaag ccgtcaattg tctgattcgt taccaattat gacaacttga cggctacatc
      121 attcactttt tcttcacaac cggcacggaa ctcgctcggg ctggccccgg tgcatttttt
      181 aaatacccgc gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat
      241 aggcatccgg gtggtgctca aaagcagctt cgcctggctg atacgttggt cctcgcgcca
      301 gcttaagacg ctaatcccta actgctggcg gaaaagatgt gacagacgcg acggcgacaa
      361 gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt gatcgctgat
      421 gtactgacaa gcctcgcgta cccgattatc catcggtgga tggagcgact cgttaatcgc
      481 ttccatgcgc cgcagtaaca attgctcaag cagatttatc gccagcagct ccgaatagcg
      541 cccttcccct tgcccggcgt taatgatttg cccaaacagg tcgctgaaat gcggctggtg
      601 cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt gacggccagt taagccattc
      661 atgccagtag gcgcgcggac gaaagtaaac ccactggtga taccattcgc gagcctccgg
      721 atgacgaccg tagtgatgaa tctctcctgg cgggaacagc aaaatatcac ccggtcggca
      781 aacaaattct cgtccctgat ttttcaccac cccctgaccg cgaatggtga gattgagaat
      841 ataacctttc attcccagcg gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat
      901 cggcgttaaa cccgccacca gatgggcatt aaacgagtat cccggcagca ggggatcatt
      961 ttgcgcttca gccatacttt tcatactccc gccattcaga gaagaaacca attgtccata
     1021 ttgcatcaga cattgccgtc actgcgtctt ttactggctc ttctcgctaa ccaaaccggt
     1081 aaccccgctt attaaaagca ttctgtaaca aagcgggacc aaagccatga caaaaacgcg
     1141 taacaaaagt gtctataatc acggcagaaa agtccacatt gattatttgc acggcgtcac
     1201 actttgctat gccatagcat ttttatccat aagattagcg gatcctacct gacgcttttt
     1261 atcgcaactc tctactgttt ctccataccc gtttttttgg gctagcagga ggaattcacc
     1321 atggtacccc ggaggttcca ccatgagcca atttgatata ttatgtaaaa caccacctaa
     1381 ggtcctggtt cgtcagtttg tggaaaggtt tgaaagacct tcaggggaaa aaatagcatc
     1441 atgtgctgct gaactaacct atttatgttg gatgattact cataacggaa cagcaatcaa
     1501 gagagccaca ttcatgagct ataatactat cataagcaat tcgctgagtt tcgatattgt
     1561 caacaaatca ctccagttta aatacaagac gcaaaaagca acaattctgg aagcctcatt
     1621 aaagaaatta attcctgctt ggggatttac aattattcct tacaatggac aaaaacatca
     1681 atctgatatc actgatattg taagtagttt gcaattacag ttcgaatcat cggaagaagc
     1741 agataaggga aatagccaca gtaaaaaaat gcttaaagca cttctaagtg agggtgaaag
     1801 catctgggag atcactgaga aaatactaaa ttcgtttgag tatacctcga gatttacaaa
     1861 aacaaaaact ttataccaat tcctcttcct agctactttc atcaattgtg gaagattcag
     1921 cgatattaag aacgttgatc cgaaatcatt taaattagtc caaaataagt atctgggagt
     1981 aataatccag tgtttagtga cagagacaaa gacaagcgtt agtaggcaca tatacttctt
     2041 tagcgcaagg ggtaggatcg atccacttgt atatttggat gaatttttga ggaattctga
     2101 accagtccta aaacgagtaa ataggaccgg caattcttca agcaacaaac aggaatacca
     2161 attattaaaa gataacttag tcagatcgta caacaaggct ttgaagaaaa atgcgcctta
     2221 tccaatcttt gctataaaga atggcccaaa atctcacatt ggaagacatt tgatgacctc
     2281 atttctgtca atgaagggcc taacggagtt gactaatgtt gtgggaaatt ggagcgataa
     2341 gcgtgcttct gccgtggcca ggacaacgta tactcatcag ataacagcaa tacctgatca
     2401 ctacttcgca ctagtttctc ggtactatgc atatgatcca atatcaaagg aaatgatagc
     2461 attgaaggat gagactaatc caattgagga gtggcagcat atagaacagc taaagggtag
     2521 tgctgaagga agcatacgat accccgcatg gaatgggata atatcacagg aggtactaga
     2581 ctacctttca tcctacataa atagacgcat ataaggtacc cggggatcct ctagagtcga
     2641 ctctagagga tcgatccttt ttaacccatc acatatacct gccgttcact attatttagt
     2701 gaaatgagat attatgatat tttctgaatt gtgattaaaa aggcaacttt atgcccatgc
     2761 aacagaaact ataaaaaata cagagaatga aaagaaacag atagattttt tagttcttta
     2821 ggcccgtagt ctgcaaatcc ttttatgatt ttctatcaaa caaaagagga aaatagacca
     2881 gttgcaatcc aaacgagagt ctaatagaat gaggtcgaaa agtaaatcgc gcgggtttgt
     2941 tactgataaa gcaggcaaga cctaaaatgt gtaaagggca aagtgtatac tttggcgtca
     3001 ccccttacat attttaggtc tttttttatt gtgcgtaact aacttgccat cttcaaacag
     3061 gagggctgga agaagcagac cgctaacaca gtacataaaa aaggagacat gaacgatgaa
     3121 catcaaaaag tttgcaaaac aagcaacagt attaaccttt actaccgcac tgctggcagg
     3181 aggcgcaact caagcgtttg cgaaagaaac gaaccaaaag ccatataagg aaacatacgg
     3241 catttcccat attacacgcc atgatatgct gcaaatccct gaacagcaaa aaaatgaaaa
     3301 atatcaagtt cctgaattcg attcgtccac aattaaaaat atctcttctg caaaaggcct
     3361 ggacgtttgg gacagctggc cattacaaaa cgctgacggc actgtcgcaa actatcacgg
     3421 ctaccacatc gtctttgcat tagccggaga tcctaaaaat gcggatgaca catcgattta
     3481 catgttctat caaaaagtcg gcgaaacttc tattgacagc tggaaaaacg ctggccgcgt
     3541 ctttaaagac agcgacaaat tcgatgcaaa tgattctatc ctaaaagacc aaacacaaga
     3601 atggtcaggt tcagccacat ttacatctga cggaaaaatc cgtttattct acactgattt
     3661 ctccggtaaa cattacggca aacaaacact gacaactgca caagttaacg tatcagcatc
     3721 agacagctct ttgaacatca acggtgtaga ggattataaa tcaatctttg acggtgacgg
     3781 aaaaacgtat caaaatgtac agcagttcat cgatgaaggc aactacagct caggcgacaa
     3841 ccatacgctg agagatcctc actacgtaga agataaaggc cacaaatact tagtatttga
     3901 agcaaacact ggaactgaag atggctacca aggcgaagaa tctttattta acaaagcata
     3961 ctatggcaaa agcacatcat tcttccgtca agaaagtcaa aaacttctgc aaagcgataa
     4021 aaaacgcacg gctgagttag caaacggcgc tctcggtatg attgagctaa acgatgatta
     4081 cacactgaaa aaagtgatga aaccgctgat tgcatctaac acagtaacag atgaaattga
     4141 acgcgcgaac gtctttaaaa tgaacggcaa atggtacctg ttcactgact cccgcggatc
     4201 aaaaatgacg attgacggca ttacgtctaa cgatatttac atgcttggtt atgtttctaa
     4261 ttctttaact ggcccataca agccgctgaa caaaactggc cttgtgttaa aaatggatct
     4321 tgatcctaac gatgtaacct ttacttactc acacttcgct gtacctcaag cgaaaggaaa
     4381 caatgtcgtg attacaagct atatgacaaa cagaggattc tacgcagaca aacaatcaac
     4441 gtttgcgcca agcttcctgc tgaacatcaa aggcaagaaa acatctgttg tcaaagacag
     4501 catccttgaa caaggacaat taacagttaa caaataagca tgcaagcttg gctgttttgg
     4561 cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat
     4621 aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
     4681 agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa
     4741 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     4801 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     4861 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     4921 aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt
     4981 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5041 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5101 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5161 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5221 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5281 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     5341 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5401 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5461 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5521 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5581 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5641 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5701 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5761 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5821 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5881 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5941 gtttactcat atatacttta gattgattta cgcgccctgt agcggcgcat taagcgcggc
     6001 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     6061 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     6121 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     6181 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     6241 gacgttggag tccacgttct ttaatagtgg actcttgttc caaacttgaa caacactcaa
     6301 ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     6361 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     6421 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6481 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6541 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6601 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6661 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     6721 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6781 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6841 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6901 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6961 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     7021 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     7081 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     7141 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     7201 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     7261 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     7321 attttctcct tacgcatctg tgcggtattt cacaccgcat agggtcatgg ctgcgccccg
     7381 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     7441 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     7501 gaaacgcgcg aggcagcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     7561 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     7621 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     7681 cgatgcgtcc ggcgtagagg atctgctcat gtttgacagc ttat
//
