LOCUS       40924_35908        8445 bp DNA     circular SYN 15-JUN-2019
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8445)
  AUTHORS   xsan
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8445)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8445
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..78)
                     /label=lacI promoter
     promoter        124..142
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    143..167
                     /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)."
     RBS             182..204
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        411..429
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    430..454
                     /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)."
     CDS             497..1678
                     /codon_start=1
                     /label=TniQ(WP_000479715.1)
                     /translation="MFLQRPKPYSDESLESFFIRVANKNGYGDVHRFLEATKRFLQDID
                     HNGYQTFPTDITRINPYSAKNSSSARTASFLKLAQLTFNEPPELLGLAINRTNMKYSPS
                     TSAVVRGAEVFPRSLLRTHSIPCCPLCLRENGYASYLWHFQGYEYCHSHNVPLITTCSC
                     GKEFDYRVSGLKGICCKCKEPITLTSRENGHEAACTVSNWLAGHESKPLPNLPKSYRWG
                     LVHWWMGIKDSEFDHFSFVQFFSNWPRSFHSIIEDEVEFNLEHAVVSTSELRLKDLLGR
                     LFFGSIRLPERNLQHNIILGELLCYLENRLWQDKGLIANLKMNALEATVMLNCSLDQIA
                     SMVEQRILKPNRKSKPNSPLDVTDYLFHFGDIFCLWLAEFQSDEFNRSFYVSRW"
     CDS             1682..3604
                     /codon_start=1
                     /label=Cas8
                     /note="accession number:  WP_001194012.1"
                     /translation="MQTLKELIASNPDDLTTELKRAFRPLTPHIAIDGNELDALTILVN
                     LTDKTDDQKDLLDRAKCKQKLRDEKWWASCINCVNYRQSHNPKFPDIRSEGVIRTQALG
                     ELPSFLLSSSKIPPYHWSYSHDSKYVNKSAFLTNEFCWDGEISCLGELLKDADHPLWNT
                     LKKLGCSQKTCKAMAKQLADITLTTINVTLAPNYLTQISLPDSDTSYISLSPVASLSMQ
                     SHFHQRLQDENRHSAITRFSRTTNMGVTAMTCGGAFRMLKSGAKFSSPPHHRLNSKRSW
                     LTSEHVQSLKQYQRLNKSLIPENSRIALRRKYKIELQNMVRSWFAMQDHTLDSNILIQH
                     LNHDLSYLGATKRFAYDPAMTKLFTELLKRELSNSINNGEQHTNGSFLVLPNIRVCGAT
                     ALSSPVTVGIPSLTAFFGFVHAFERNINRTTSSFRVESFAICVHQLHVEKRGLTAEFVE
                     KGDGTISAPATRDDWQCDVVFSLILNTNFAQHIDQDTLVTSLPKRLARGSAKIAIDDFK
                     HINSFSTLETAIESLPIEAGRWLSLYAQSNNNLSDLLAAMTEDHQLMASCVGYHLLEEP
                     KDKPNSLRGYKHAIAECIIGLINSITFSSETDPNTIFWSLKNYQNYLVVQPRSINDETT
                     DKSSL"
     CDS             3576..4634
                     /codon_start=1
                     /label=Cas7
                     /note="accession number: WP_000775488.1"
                     /translation="MKLPTNLAYERSIDPSDVCFFVVWPDDRKTPLTYNSRTLLGQMEA
                     ASLAYDVSGQPIKSATAEALAQGNPHQVDFCHVPYGASHIECSFSVSFSSELRQPYKCN
                     SSKVKQTLVQLVELYETKIGWTELATRYLMNICNGKWLWKNTRKAYCWNIVLTPWPWNG
                     EKVGFEDIRTNYTSRQDFKNNKNWSAIVEMIKTAFSSTDGLAIFEVRATLHLPTNAMVR
                     PSQVFTEKESGSKSKSKTQNSRVFQSTTIDGERSPILGAFKTGAAIATIDDWYPEATEP
                     LRVGRFGVHREDVTCYRHPSTGKDFFSILQQAEHYIEVLSANKTPAQETINDMHFLMAN
                     LIKGGMFQHKGD"
     CDS             4868..5236
                     /codon_start=1
                     /product="type I-F CRISPR-associated endoribonuclease
                     Cas6/Csy4 [Vibrio cholerae]"
                     /label=Cas6
                     /note="accession number: WP_000872476.1"
                     /translation="MEQLQYFHISNTVLVPEDCTYVSFRRCQSIDKLTAAGLARKIRRL
                     EKRALSRGEQFDPSSFAQKEHTAIAHYHSLGESSKQTNRNFRLNIRMLSEQPREGNSIF
                     SSYGLSNSENSFQPVPLI"
     terminator      5266..5313
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5487..6275)
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(6276..6367)
                     /label=AmpR promoter
     rep_origin      complement(6415..7153)
                     /direction=LEFT
                     /label=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."
     protein_bind    complement(7329..7350)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(7366..8445)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
       61 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
      121 aattaatacg actcactata ggggaattgt gagcggataa caattcccct gtagaaataa
      181 ttttgtttaa ctttaataag gagatatacc atgggtgaac tgccgagtag gtagctgata
      241 acgttgtctg acacttgtca caaaccgcta ggaggtgaac tgccgagtag gtagctgata
      301 acggatccga attcgagctc ggcgcgcctg caggtcgaca agcttgcggc cgcataatgc
      361 ttaagtcgaa cagaaagtaa tcgtattgta cacggccgca taatcgaaat taatacgact
      421 cactataggg gaattgtgag cggataacaa ttccccatct tagtatatta gttaagtata
      481 agaaggagat atacatatgt ttttgcaaag acctaaacct tacagcgatg aaagtttaga
      541 aagtttcttt atccgagtgg ctaacaaaaa tggctacggt gatgtccatc gcttcctaga
      601 agccactaaa cgattccttc aagacattga ccataatggc tatcaaacct ttccgactga
      661 tataactcgg ataaacccat actcagctaa aaacagttcc agcgcacgaa ctgcgtcatt
      721 cctgaaactt gcacaattga catttaatga accgccagag ctacttgggt tggcaattaa
      781 cagaacaaac atgaaatact cgccgtcaac tagcgcggtt gttagaggtg cagaagtctt
      841 tcctcgcagt ttactacgga cgcactccat cccctgctgt cctttgtgtc tgcgagaaaa
      901 tggctacgcc tcctaccttt ggcactttca ggggtacgaa tactgccaca gccataacgt
      961 acctttaatt accacttgta gctgtggtaa ggagtttgac taccgagtat ctgggttaaa
     1021 gggcatttgc tgcaaatgca aggagcctat caccttaacc agcagggaga acggtcatga
     1081 ggcagcgtgt actgtttcaa actggcttgc tggccatgaa tctaaacctc tgccaaatct
     1141 tcctaaaagc taccgatggg gtttagttca ttggtggatg ggtattaaag atagcgagtt
     1201 cgatcacttt tcgttcgttc aatttttctc aaactggcca aggtcattcc actcgataat
     1261 cgaagatgaa gtagagttca accttgagca tgctgttgtc agcacgtctg aattacgact
     1321 aaaagatctt cttggtcgat tgtttttcgg ttcaattcgg ttacctgagc ggaatcttca
     1381 acacaatatc atccttggtg agcttctctg ctatttagaa aatcgtttat ggcaagacaa
     1441 gggattaatc gccaacctca aaatgaacgc gttagaggcg actgtaatgt taaattgtag
     1501 cctcgatcag attgcatcaa tggttgaaca acgcatcttg aagccaaatc gaaaaagcaa
     1561 gcccaacagc cctcttgatg ttaccgatta tctatttcat ttcggcgata ttttctgtct
     1621 ttggttagct gagttccaaa gcgatgagtt taaccgttcg ttttatgtgt cgaggtggta
     1681 aatgcaaact ctgaaagaac taatcgcatc caatcctgac gacttaacaa ctgaacttaa
     1741 gagagcattt cgtccactca caccgcatat tgcaattgat ggtaatgaac ttgacgcact
     1801 gacgatatta gtcaatttaa ccgataagac tgatgatcag aaagacctgc tcgatcgagc
     1861 caaatgcaag caaaaacttc gagatgaaaa atggtgggct agctgcataa attgcgttaa
     1921 ttacagacaa agccataacc caaaattccc ggatatacgt tctgaaggcg tgattcgaac
     1981 ccaagccctg ggtgaattac cgagtttcct actctcgtcc tcaaaaatcc caccatacca
     2041 ttggtcatat agccatgatt caaaatacgt caacaaaagc gcattcctca ccaatgagtt
     2101 ttgttgggat ggtgagatct catgtttggg tgagcttctt aaagatgcag atcacccact
     2161 ttggaatact ttgaaaaagt taggttgttc tcaaaaaact tgcaaagcaa tggcaaaaca
     2221 actagctgat attactctca cgactatcaa tgtcacactc gcaccaaatt acctgactca
     2281 aatatctctc cccgatagtg atacatccta catttcactc tctcctgttg catcgctatc
     2341 gatgcaaagc cactttcatc agaggctcca agatgaaaat cggcatagcg cgataacgcg
     2401 gtttagccga actaccaaca tgggggtcac tgcgatgaca tgtggcggcg catttagaat
     2461 gttaaagtct ggcgctaagt tttccagccc ccctcatcac cgattaaaca gtaaacgaag
     2521 ttggttgacg tcagagcatg ttcagtcatt aaaacagtac cagcgcctca acaaaagcct
     2581 catacctgaa aactctcgga ttgcactccg tagaaaatac aaaatcgagc ttcaaaatat
     2641 ggtcagatct tggtttgcaa tgcaagacca tacccttgat tcgaacatac ttatccaaca
     2701 tttgaatcat gacctatctt acttaggagc cacaaaacgt tttgcatacg atccagcgat
     2761 gaccaagctc tttactgagc ttttgaaacg agagttatca aattcaatca ataatggtga
     2821 gcaacacact aatggatcgt ttttagtcct accgaatatc agggtttgtg gcgcaacagc
     2881 tttaagctcc ccggtaacgg tggggattcc atcacttaca gctttctttg gcttcgttca
     2941 cgcatttgaa cggaatataa atcgcaccac ctcatcgttt cgtgttgaat cctttgcgat
     3001 atgcgtccat caactacatg tcgagaagcg aggtttgaca gcagagtttg tggaaaaagg
     3061 cgacgggact atatccgctc ccgcgacccg ggatgactgg cagtgtgatg tcgtatttag
     3121 ccttattttg aacaccaact ttgctcaaca tattgaccaa gatacgttag ttacatcact
     3181 accaaagcga ttggctcggg gttcagcaaa aattgcgatt gatgacttta aacatatcaa
     3241 ctcattctcg acattagaaa cagcgatcga atctctgcca atagaagctg gtaggtggtt
     3301 atcactttac gcacagtcaa acaataatct aagtgatcta ttagcagcca tgacagagga
     3361 ccatcagctc atggcaagct gcgtcggtta ccacttgtta gaagagccca aagataaacc
     3421 aaactccctc agaggttaca aacacgctat cgccgagtgc atcattggac tcattaactc
     3481 aatcaccttt agctcagaga ctgatcccaa cacaatcttt tggtcgctaa agaactatca
     3541 aaactaccta gtggtacagc caaggagtat caacgatgaa actaccgaca aatctagcct
     3601 atgagcgctc tatcgaccca tcagatgtct gtttttttgt cgtctggccc gatgatagaa
     3661 aaacaccttt aacctacaat tctcgtactc tgctcgggca aatggaagcg gcatcattag
     3721 cctatgatgt ctcaggtcaa ccaataaaaa gtgccaccgc tgaggcgtta gctcaaggga
     3781 accctcatca agttgatttc tgccacgttc catacggtgc gagtcatatt gaatgcagtt
     3841 tctccgtctc gttttcttct gaactacgtc aaccatataa gtgtaactca agcaaagtta
     3901 aacaaacgct agtgcaatta gtcgagctct acgaaacgaa aatcggctgg actgagctag
     3961 caacccgata tttgatgaac atttgcaacg gtaaatggct gtggaaaaat acccgtaaag
     4021 cctattgctg gaacattgta cttacacctt ggccttggaa cggggaaaag gttggatttg
     4081 aagatattcg tactaactac acctcacggc aagactttaa aaataataaa aattggtctg
     4141 ctatagttga aatgatcaaa accgcatttt ctagtactga tgggctggcg atatttgaag
     4201 tcagggccac cttgcacttg ccaacgaatg ctatggtgcg gccaagccaa gttttcacag
     4261 aaaaagaaag tggcagtaaa agtaaatcta aaactcaaaa cagtcgagtt tttcagagta
     4321 caactattga tggtgaacga tcgccaatac taggggcctt taaaacggga gcagctattg
     4381 caaccattga cgactggtat cctgaagcca ctgagccact aagggtcgga cggtttgggg
     4441 ttcatcgcga agatgtcact tgctaccgtc atccgtctac cggaaaagat tttttctcga
     4501 tattacaaca agcagagcac tatattgaag tgttgagcgc caacaaaact cccgctcaag
     4561 aaactatcaa cgacatgcac tttttaatgg ctaacctgat taagggtggg atgttccagc
     4621 ataaaggaga ctgactgtga aatggtatta taagacaatc acctttctgc cagagttgtg
     4681 caacaacgag tcactggctg caaagtgtct ccgcgttctg catggattta actatcagta
     4741 tgagacacga aatatcggcg tttcatttcc gctttggtgt gatgcaacgg ttggaaaaaa
     4801 gatttcattt gtcagcaaga acaagataga actcgactta ctacttaaac aacactattt
     4861 cgtccaaatg gaacaacttc aatactttca tatatccaac actgttctcg tcccagaaga
     4921 ttgtacatac gtttccttta gacgctgtca atctatagat aagctcacag cagcagggct
     4981 ggcaaggaaa atcagacgcc tggagaaacg tgctctatct agaggcgagc aatttgaccc
     5041 atcatctttt gctcaaaaag agcatactgc aatagcgcac taccactcac ttggggagtc
     5101 cagcaaacag acgaaccgca actttcgact caatatcagg atgctctcgg agcaaccccg
     5161 tgaggggaac tcgattttta gtagctatgg cttatcaaat tcagaaaact cgtttcagcc
     5221 tgtaccctta atctgaccta ggctgctgcc accgctgagc aataactagc ataacccctt
     5281 ggggcctcta aacgggtctt gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac
     5341 acggtcacac tgcttccggt agtcaataaa ccggtaaacc agcaatagac ataagcggct
     5401 atttaacgac cctgccctga accgacgacc gggtcatcgt ggccggatct tgcggcccct
     5461 cggcttgaac gaattgttag acattatttg ccgactacct tggtgatctc gcctttcacg
     5521 tagtggacaa attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca
     5581 agataagcct gtctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt
     5641 gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg
     5701 cgggacaacg taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat
     5761 agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt
     5821 tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata
     5881 gccagatcaa tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc
     5941 cattctccaa attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc
     6001 acaacaatgg tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt
     6061 tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta
     6121 accagcaaat caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa
     6181 tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt
     6241 tgagtcgata cttcggcgat caccgcttcc ctcatactct tcctttttca atattattga
     6301 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6361 aaacaaatag ctagctcact cggtcgctac gctccgggcg tgagactgcg gcgggcgctg
     6421 cggacacata caaagttacc cacagattcc gtggataagc aggggactaa catgtgaggc
     6481 aaaacagcag ggccgcgccg gtggcgtttt tccataggct ccgccctcct gccagagttc
     6541 acataaacag acgcttttcc ggtgcatctg tgggagccgt gaggctcaac catgaatctg
     6601 acagtacggg cgaaacccga caggacttaa agatccccac cgtttccggc gggtcgctcc
     6661 ctcttgcgct ctcctgttcc gaccctgccg tttaccggat acctgttccg cctttctccc
     6721 ttacgggaag tgtggcgctt tctcatagct cacacactgg tatctcggct cggtgtaggt
     6781 cgttcgctcc aagctgggct gtaagcaaga actccccgtt cagcccgact gctgcgcctt
     6841 atccggtaac tgttcacttg agtccaaccc ggaaaagcac ggtaaaacgc cactggcagc
     6901 agccattggt aactgggagt tcgcagagga tttgtttagc taaacacgcg gttgctcttg
     6961 aagtgtgcgc caaagtccgg ctacactgga aggacagatt tggttgctgt gctctgcgaa
     7021 agccagttac cacggttaag cagttcccca actgacttaa ccttcgatca aaccacctcc
     7081 ccaggtggtt ttttcgttta cagggcaaaa gattacgcgc agaaaaaaag gatctcaaga
     7141 agatcctttg atcttttcta ctgaaccgct ctagatttca gtgcaattta tctcttcaaa
     7201 tgtagcacct gaagtcagcc ccatacgata taagttgtaa ttctcatgtt agtcatgccc
     7261 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc
     7321 ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     7381 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     7441 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     7501 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     7561 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     7621 atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc
     7681 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     7741 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     7801 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     7861 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     7921 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     7981 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     8041 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     8101 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     8161 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     8221 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     8281 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     8341 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     8401 gacaccggca tactctgcga catcgtataa cgttactggt ttcac
//
