LOCUS       Exported                6632 bp ds-DNA     circular SYN 06-MAR-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCDF-casBCDE(+6)
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6632)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Mar 6, 2017 from SnapGene 2.6.2
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6632
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     protein_bind    3..27
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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)."
     CDS             80..103
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin, an engineered 
                     form of streptavidin"
                     /note="Strep-Tag II"
                     /translation="WSHPQFEK"
     CDS             110..133
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /note="HRV 3C site"
                     /translation="LEVLFQGP"
     CDS             143..625
                     /codon_start=1
                     /note="casB"
                     /translation="MADEIDAMALYRAWQQLDNGSCAQIRRVSEPDELRDIPAFYRLVQ
                     PFGWENPRHQQALLRMVFCLSAGKNVIRHQDKKSEQTTGISLGRALANSGRINERRIFQ
                     LIRADRTADMVQLRRLLTHAEPVLDWPLMARMLTWWGKRERQQLLEDFVLTTNKNA"
     CDS             638..1729
                     /codon_start=1
                     /note="casC"
                     /translation="MSNFINIHVLISHSPSCLNRDDMNMQKDAIFGGKRRVRISSQSLK
                     RAMRKSGYYAQNIGESSLRTIHLAQLRDVLRQKLGERFDQKIIDKTLALLSGKSVDEAE
                     KISADAVTPWVVGEIAWFCEQVAKAEADNLDDKKLLKVLKEDIAAIRVNLQQGVDIALS
                     GRMATSGMMTELGKVDGAMSIAHAITTHQVDSDIDWFTAVDDLQEQGSAHLGTQEFSSG
                     VFYRYANINLAQLQENLGGASREQALEIATHVVHMLATEVPGAKQRTYAAFNPADMVMV
                     NFSDMPLSMANAFEKAVKAKDGFLQPSIQAFNQYWDRVANGYGLNGAAAQFSLSDVDPI
                     TAQVKQMPTLEQLKSWVRNNGEA"
     CDS             1732..2406
                     /codon_start=1
                     /note="casD"
                     /translation="MRSYLILRLAGPMQAWGQPTFEGTRPTGRFPTRSGLLGLLGACLG
                     IQRDDTSSLQALSESVQFAVRCDELILDDRRVSVTGLRDYHTVLGAREDYRGLKSHETI
                     QTWREYLCDASFTVALWLTPHATMVISELEKAVLKPRYTPYLGRRSCPLTHPLFLGTCQ
                     ASDPQKALLNYEPVGGDIYSEESVTGHHLKFTARDEPMITLPRQFASREWYVIKGGMDV
                     SQ"
     CDS             2393..2992
                     /codon_start=1
                     /note="casE"
                     /translation="MYLSKVIIARAWSRDLYQLHQGLWHLFPNRPDAARDFLFHVEKRN
                     TPEGCHVLLQSAQMPVSTAVATVIKTKQVEFQLQVGVPLYFRLRANPIKTILDNQKRLD
                     SKGNIKRCRVPLIKEAEQIAWLQRKLGNAARVEDVHPISERPQYFSGDGKSGKIQTVCF
                     EGVLTINDAPALIDLVQQGIGPAKSMGCGLLSLAPL"
     repeat_region   3154..3181
                     /note="repeat"
                     /note="repeat from E.coli K12  type I-E CRISPR I array"
     ncRNA           3182..3220
                     /note="spacer_g8+6"
                     /ncRNA_class="guide_RNA"
     repeat_region   3221..3248
                     /note="repeat"
                     /note="repeat from E.coli K12  type I-E CRISPR I array"
     terminator      3313..3360
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3531..4322)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /note="SmR"
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(4323..4414)
                     /gene="bla"
                     /note="AmpR promoter"
     rep_origin      complement(4462..5200)
                     /direction=LEFT
                     /note="CloDF13 ori"
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(5410..6492)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /note="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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(6493..6570)
                     /gene="lacI"
                     /note="lacI promoter"
     promoter        6616..2
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggcaagct ggagccaccc gcagttcgaa aagggtgcac ttgaagtcct
      121 ctttcaggga cccgggtacc agatggctga tgaaattgat gcaatggctt tatatcgagc
      181 ctggcaacaa ctggataatg gatcatgtgc gcaaattaga cgtgtttcag aacctgatga
      241 attacgcgat atccctgcgt tttataggct ggtgcaacct tttggttggg aaaacccacg
      301 tcaccagcag gctcttttgc gcatggtgtt ttgcctgagc gcaggaaaga atgtcatccg
      361 acatcaggac aaaaaatcgg agcaaacaac aggtatctcg ttgggaagag ctttagccaa
      421 tagtggaaga attaacgagc gccgtatctt tcaattaatt cgggctgaca gaacagccga
      481 tatggtccag ttacgtcgat tacttactca cgccgaaccc gtacttgact ggccattaat
      541 ggccaggatg ttgacctggt ggggaaagcg cgaacgccag caacttctgg aagattttgt
      601 attgaccaca aacaaaaatg cgtaaggaaa cctttctatg tctaacttta tcaatattca
      661 tgttctgatc tctcacagcc cttcatgtct gaaccgcgac gatatgaaca tgcagaaaga
      721 cgctattttc ggcggcaaaa gacgagtaag aatttcaagt caaagcctta aacgtgcgat
      781 gcgtaaaagt ggttattacg cacaaaatat tggtgaatcc agtctcagaa ccattcatct
      841 tgcacaatta cgtgatgttc ttcggcaaaa acttggtgaa cgttttgacc aaaaaatcat
      901 cgataagaca ttagcgctgc tctccggtaa atcagttgat gaagccgaaa agatttctgc
      961 cgatgcggtt actccctggg ttgtgggaga aatagcctgg ttctgtgagc aggttgcaaa
     1021 agcagaggct gataatctgg atgataaaaa gctgctcaaa gttcttaagg aagatattgc
     1081 cgccatacgt gtgaatttac agcagggtgt tgatattgcg cttagtggaa gaatggcaac
     1141 cagcggcatg atgactgagt tgggaaaagt tgatggtgca atgtccattg cgcatgcgat
     1201 cactactcat caggttgatt ctgatattga ctggttcacc gctgtagatg atttacagga
     1261 acaaggttct gcacatctgg gaactcagga attttcatcg ggtgtttttt atcgttatgc
     1321 caacattaac ctcgctcaac ttcaggaaaa tttaggtggt gcctccaggg agcaggctct
     1381 ggaaattgca acccatgttg ttcatatgct ggcaacagag gtccctggag caaaacagcg
     1441 tacttatgcc gcttttaacc ctgcggatat ggtaatggtt aatttctccg atatgccact
     1501 ttctatggca aatgcttttg aaaaagcggt taaagcgaaa gatggctttt tgcaaccgtc
     1561 tatacaggcg tttaatcaat attgggatcg cgttgccaat ggatatggtc tgaacggagc
     1621 tgctgcgcaa ttcagcttat ctgatgtaga cccaattact gctcaagtta aacaaatgcc
     1681 tactttagaa cagttaaaat cctgggttcg taataatggc gaggcgtgaa catgagatct
     1741 tatttgatct tgcggcttgc tgggccaatg caagcctggg ggcagccgac ctttgaagga
     1801 acgcgaccta ccggaagatt tccgacccga agcgggttat tagggctact cggggcttgt
     1861 cttgggatcc aacgtgatga tacttcttca ttacaggcgt tatcagagag tgtgcaattt
     1921 gcagtgcgct gcgatgaact cattcttgac gatcgtcgtg tgtctgtaac ggggttgcgt
     1981 gattaccata cagtccttgg agcgcgagaa gattaccgtg gtttgaaaag tcatgaaacg
     2041 attcaaacat ggcgcgaata tttatgtgat gcctccttta ccgtcgctct ctggttaaca
     2101 ccccatgcaa cgatggttat ctcagaactt gaaaaagcag tattaaagcc tcggtataca
     2161 ccttacctgg ggcggagaag ttgcccacta acacacccgc tttttttggg gacatgtcag
     2221 gcatcggatc ctcagaaggc gctattaaat tatgagcccg ttggcggcga tatatatagt
     2281 gaggaatcag ttacagggca tcatttaaaa tttacggcgc gcgacgaacc gatgatcacc
     2341 ttgcctcgac aatttgcttc ccgagaatgg tatgtgatta aaggaggtat ggatgtatct
     2401 cagtaaagtc atcattgcca gggcctggag cagggatctt taccaacttc accagggatt
     2461 atggcattta tttccaaaca gaccggatgc tgctcgtgat tttctttttc atgttgagaa
     2521 gcgaaacaca ccagaaggct gtcatgtttt attgcagtca gcgcaaatgc ctgtttcaac
     2581 tgccgttgcg acagtcatta aaactaaaca ggttgaattt caacttcagg ttggtgttcc
     2641 actctatttt cggcttcggg caaatccgat caaaactatt ctcgacaatc aaaagcgcct
     2701 ggacagtaaa gggaatatta aacgctgtcg ggttccgtta ataaaagaag cagaacaaat
     2761 cgcgtggttg caacgtaaat tgggcaatgc ggcgcgcgtt gaagatgtgc atcccatatc
     2821 ggaacggcca cagtattttt ctggtgatgg taaaagtgga aagatccaaa cggtttgctt
     2881 tgaaggtgtg ctcaccatca acgacgcgcc agcgttaata gatcttgtac agcaaggtat
     2941 tgggccagct aaatcgatgg gatgtggctt gctatctttg gctccactgt gagcggccgc
     3001 actcgagatt tggatggttt aaggttggtg tcttttttac ctgtttgaaa acaaagaatt
     3061 agctgatctt taataataag gaaatgttac attaaggttg gtgggttgtt tttatgggaa
     3121 aaaatgcttt aagaacaaat gtatactttt agagagttcc ccgcgccagc ggggataaac
     3181 cgctgtcttt cgctgctgag ggtgacgatc ccgcatgtat gagttccccg cgccagcggg
     3241 gataaaccga gctcccattt tcaaacccat caagacgcct tcgcctaggc tgctgccacc
     3301 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     3361 ctgaaacctc aggcatttga gaagcacacg gtcacactgc ttccggtagt caataaaccg
     3421 gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc gacgaccggg
     3481 tcatcgtggc cggatcttgc ggcccctcgg cttgaacgaa ttgttagaca ttatttgccg
     3541 actaccttgg tgatctcgcc tttcacgtag tggacaaatt cttccaactg atctgcgcgc
     3601 gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag tatgacgggc
     3661 tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt cggcgcgatt
     3721 ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg
     3781 ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc ctcaaataga
     3841 tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa ggcaacgcta
     3901 tgttctcttg cttttgtcag caagatagcc agatcaatgt cgatcgtggc tggctcgaag
     3961 atacctgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg cttagctgga
     4021 taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc gcggagaatc
     4081 tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt
     4141 gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg tggcttcagg
     4201 ccgccatcca ctgcggagcc gtacaaatgt acggccagca acgtcggttc gagatggcgc
     4261 tcgatgacgc caactacctc tgatagttga gtcgatactt cggcgatcac cgcttccctc
     4321 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4381 tacatatttg aatgtattta gaaaaataaa caaatagcta gctcactcgg tcgctacgct
     4441 ccgggcgtga gactgcggcg ggcgctgcgg acacatacaa agttacccac agattccgtg
     4501 gataagcagg ggactaacat gtgaggcaaa acagcagggc cgcgccggtg gcgtttttcc
     4561 ataggctccg ccctcctgcc agagttcaca taaacagacg cttttccggt gcatctgtgg
     4621 gagccgtgag gctcaaccat gaatctgaca gtacgggcga aacccgacag gacttaaaga
     4681 tccccaccgt ttccggcggg tcgctccctc ttgcgctctc ctgttccgac cctgccgttt
     4741 accggatacc tgttccgcct ttctccctta cgggaagtgt ggcgctttct catagctcac
     4801 acactggtat ctcggctcgg tgtaggtcgt tcgctccaag ctgggctgta agcaagaact
     4861 ccccgttcag cccgactgct gcgccttatc cggtaactgt tcacttgagt ccaacccgga
     4921 aaagcacggt aaaacgccac tggcagcagc cattggtaac tgggagttcg cagaggattt
     4981 gtttagctaa acacgcggtt gctcttgaag tgtgcgccaa agtccggcta cactggaagg
     5041 acagatttgg ttgctgtgct ctgcgaaagc cagttaccac ggttaagcag ttccccaact
     5101 gacttaacct tcgatcaaac cacctcccca ggtggttttt tcgtttacag ggcaaaagat
     5161 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctactg aaccgctcta
     5221 gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa
     5281 gttgtaattc tcatgttagt catgccccgc gcccaccgga aggagctgac tgggttgaag
     5341 gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt
     5401 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5461 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
     5521 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
     5581 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
     5641 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc
     5701 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
     5761 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
     5821 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
     5881 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
     5941 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
     6001 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
     6061 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
     6121 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
     6181 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
     6241 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
     6301 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
     6361 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     6421 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     6481 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
     6541 gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact
     6601 cctgcattag gaaattaata cgactcacta ta
//
