LOCUS Exported 8130 bp ds-DNA circular SYN 01-NOV-2017 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8130) AUTHORS . TITLE Direct Submission JOURNAL Exported Nov 1, 2017 from SnapGene 4.0.5 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8130 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 482..576 /label=lambda t0 terminator /note="/ApEinfo_fwdcolor=#c6c9d1 /ApEinfo_revcolor=#c6c9d1" CDS complement(597..1256) /label=CmR /note="/ApEinfo_fwdcolor=#b7e6d7 /ApEinfo_revcolor=#b7e6d7" terminator 1440..1483 /label=bacterial terminator /note="/ApEinfo_fwdcolor=#c6c9d1 /ApEinfo_revcolor=#c6c9d1" misc_feature complement(1468..1493) /label=SB-prep-2Ea /note="/ApEinfo_fwdcolor=#85dae9 /ApEinfo_revcolor=#85dae9" misc_feature 1486..1568 /label=geneblock prefix /note="/ApEinfo_fwdcolor=#9eafd2 /ApEinfo_revcolor=#9eafd2" misc_feature 1486..1507 /label=BioBrick prefix /note="/ApEinfo_fwdcolor=#84b0dc /ApEinfo_revcolor=#84b0dc" misc_feature 1533..5639 /label=Cas9 /note="/ApEinfo_fwdcolor=#f58a5e /ApEinfo_revcolor=#f58a5e" misc_feature 1549..2218 /label=PCR1 /note="/ApEinfo_fwdcolor=#ffef86 /ApEinfo_revcolor=#ffef86" misc_feature 2199..2991 /label=geneblock EcorI/I473F /note="/ApEinfo_fwdcolor=#c6c9d1 /ApEinfo_revcolor=#c6c9d1" misc_feature 2872..2877 /label=EcoRI /note="/ApEinfo_fwdcolor=#9eafd2 /ApEinfo_revcolor=#9eafd2" misc_feature 2949..2951 /label=I473F ATT->TTT /note="/ApEinfo_fwdcolor=#b1ff67 /ApEinfo_revcolor=#b1ff67" misc_feature 2972..7236 /label=PCR2 /note="/ApEinfo_fwdcolor=#b1ff67 /ApEinfo_revcolor=#b1ff67" misc_feature 5639..6508 /label=CRISPR-associated endonuclease Cas1 /note="/ApEinfo_fwdcolor=#c6c9d1 /ApEinfo_revcolor=#c6c9d1" misc_feature 6505..6846 /label=CRISPR-associated endoribonuclease Cas2 /note="/ApEinfo_fwdcolor=#f8d3a9 /ApEinfo_revcolor=#f8d3a9" misc_feature 6836..7498 /label=CRISPR-associated protein Csn2 /note="/ApEinfo_fwdcolor=#ff9ccd /ApEinfo_revcolor=#ff9ccd" misc_feature 7214..7588 /label=Geneblock XbaI /note="/ApEinfo_fwdcolor=#d59687 /ApEinfo_revcolor=#d59687" misc_feature 7279..7284 /label=XbaI CTA->CTT /note="/ApEinfo_fwdcolor=#b7e6d7 /ApEinfo_revcolor=#b7e6d7" misc_feature 7538..7543 /label=-35 /note="/ApEinfo_fwdcolor=#c7b0e3 /ApEinfo_revcolor=#c7b0e3" misc_feature 7559..7567 /label=-10 /note="/ApEinfo_fwdcolor=#f8d3a9 /ApEinfo_revcolor=#f8d3a9" misc_feature 7568..7588 /label=BioBrick suffix /note="/ApEinfo_fwdcolor=#84b0dc /ApEinfo_revcolor=#84b0dc" misc_feature 7579..7600 /label=SB-prep-3P-1 /note="/ApEinfo_fwdcolor=#f58a5e /ApEinfo_revcolor=#f58a5e" terminator 7589..7646 /label=his operon terminator /note="/ApEinfo_fwdcolor=#c6c9d1 /ApEinfo_revcolor=#c6c9d1" rep_origin complement(7841..299) /direction=LEFT /label=ori /note="/ApEinfo_fwdcolor=#ffef86 /ApEinfo_revcolor=#ffef86" ORIGIN 1 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 61 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 121 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga 181 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 241 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 301 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 361 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 421 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagc tcgaggcttg 481 gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga 541 gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctcgat atcaaattac 601 gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg 661 aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct 721 tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg 781 tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca 841 ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat 901 atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca 961 gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg 1021 tctttcattg ccatacgaaa ttccggatga gcattcatca ggcgggcaag aatgtgaata 1081 aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc 1141 agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct 1201 ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta 1261 gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt 1321 tcattatggt gaaagttgga acctcttacg tgcccgatca actcgagtgc cacctgacgt 1381 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggcagaa 1441 tttcagataa aaaaaatcct tagctttcgc taaggatgat ttctggaatt cgcggccgct 1501 tctagagaaa ctttttattt taggaggcaa aaatggataa gaaatactca ataggcttag 1561 atatcggcac aaatagcgtc ggatgggcgg tgatcactga tgattataag gttccgtcta 1621 aaaagttcaa ggttctggga aatacagacc gccacagtat caaaaaaaat cttatagggg 1681 ctcttttatt tgacagtgga gagacagcgg aagcgactcg tctcaaacgg acagctcgta 1741 gaaggtatac acgtcggaag aatcgtattt gttatctaca ggagattttt tcaaatgaga 1801 tggcgaaagt agatgatagt ttctttcatc gacttgaaga gtcttttttg gtggaagaag 1861 acaagaagca tgaacgtcat cctatttttg gaaatatagt agatgaagtt gcttatcatg 1921 agaaatatcc aactatctat catctgcgaa aaaaattggt agattctact gataaagcgg 1981 atttgcgctt aatctatttg gccttagcgc atatgattaa gtttcgtggt cattttttga 2041 ttgagggaga tttaaatcct gataatagtg atgtggacaa actatttatc cagttggtac 2101 aaacctacaa tcaattattt gaagaaaacc ctattaacgc aagtggagta gatgctaaag 2161 cgattctttc tgcacgattg agtaaatcaa gacgattaga aaatctcatt gctcagctcc 2221 ccggtgagaa gaaaaatggc ttatttggga atctcattgc tttgtcattg ggtttgaccc 2281 ctaattttaa atcaaatttt gatttggcag aagatgctaa attacagctt tcaaaagata 2341 cttacgatga tgatttagat aatttattgg cgcaaattgg agatcaatat gctgatttgt 2401 ttttggcagc taagaattta tcagatgcta ttttactttc agatatccta agagtaaata 2461 ctgaaataac taaggctccc ctatcagctt caatgattaa acgctacgat gaacatcatc 2521 aagacttgac tcttttaaaa gctttagttc gacaacaact tccagaaaag tataaagaaa 2581 tcttttttga tcaatcaaaa aacggatatg caggttatat tgatggggga gctagccaag 2641 aagaatttta taaatttatc aaaccaattt tagaaaaaat ggatggtact gaggaattat 2701 tggtgaaact aaatcgtgaa gatttgctgc gcaagcaacg gacctttgac aacggctcta 2761 ttccccatca aattcacttg ggtgagctgc atgctatttt gagaagacaa gaagactttt 2821 atccattttt aaaagacaat cgtgagaaga ttgaaaaaat cttgactttt cgaatacctt 2881 attatgttgg tccattggcg cgtggcaata gtcgttttgc atggatgact cggaagtctg 2941 aagaaacatt taccccatgg aattttgaag aagttgtcga taaaggtgct tcagctcaat 3001 catttattga acgcatgaca aactttgata aaaatcttcc aaatgaaaaa gtactaccaa 3061 aacatagttt gctttatgag tattttacgg tttataacga attgacaaag gtcaaatatg 3121 ttactgaagg aatgcgaaaa ccagcatttc tttcaggtga acagaagaaa gccattgttg 3181 atttactctt caaaacaaat cgaaaagtaa ccgttaagca attaaaagaa gattatttca 3241 aaaaaataga atgttttgat agtgttgaaa tttcaggagt tgaagataga tttaatgctt 3301 cattaggtac ctaccatgat ttgctaaaaa ttattaaaga taaagatttt ttggataatg 3361 aagaaaatga agatatctta gaggatattg ttttaacatt gaccttattt gaagataggg 3421 agatgattga ggaaagactt aaaacatatg ctcacctctt tgatgataag gtgatgaaac 3481 agcttaaacg tcgccgttat actggttggg gacgtttgtc tcgaaaattg attaatggta 3541 ttagggataa gcaatctggc aaaacaatat tagatttttt gaaatcagat ggttttgcca 3601 atcgcaattt tatgcagctg atccatgatg atagtttgac atttaaagaa gacattcaaa 3661 aagcacaagt gtctggacaa ggcgatagtt tacatgaaca tattgcaaat ttagctggta 3721 gccctgctat taaaaaaggt attttacaga ctgtaaaagt tgttgatgaa ttggtcaaag 3781 taatggggcg gcataagcca gaaaatatcg ttattgaaat ggcacgtgaa aatcagacaa 3841 ctcaaaaggg ccagaaaaat tcgcgagagc gtatgaaacg aatcgaagaa ggtatcaaag 3901 aattaggaag tcagattctt aaagagcatc ctgttgaaaa tactcaattg caaaatgaaa 3961 agctctatct ctattatctc caaaatggaa gagacatgta tgtggaccaa gaattagata 4021 ttaatcgttt aagtgattat gatgtcgatc acattgttcc acaaagtttc cttaaagacg 4081 attcaataga caataaggtc ttaacgcgtt ctgataaaaa tcgtggtaaa tcggataacg 4141 ttccaagtga agaagtagtc aaaaagatga aaaactattg gagacaactt ctaaacgcca 4201 agttaatcac tcaacgtaag tttgataatt taacgaaagc tgaacgtgga ggtttgagtg 4261 aacttgataa agctggtttt atcaaacgcc aattggttga aactcgccaa atcactaagc 4321 atgtggcaca aattttggat agtcgcatga atactaaata cgatgaaaat gataaactta 4381 ttcgagaggt taaagtgatt accttaaaat ctaaattagt ttctgacttc cgaaaagatt 4441 tccaattcta taaagtacgt gagattaaca attaccatca tgcccatgat gcgtatctaa 4501 atgccgtcgt tggaactgct ttgattaaga aatatccaaa acttgaatcg gagtttgtct 4561 atggtgatta taaagtttat gatgttcgta aaatgattgc taagtctgag caagaaatag 4621 gcaaagcaac cgcaaaatat ttcttttact ctaatatcat gaacttcttc aaaacagaaa 4681 ttacacttgc aaatggagag attcgcaaac gccctctaat cgaaactaat ggggaaactg 4741 gagaaattgt ctgggataaa gggcgagatt ttgccacagt gcgcaaagta ttgtccatgc 4801 cccaagtcaa tattgtcaag aaaacagaag tacagacagg cggattctcc aaggagtcaa 4861 ttttaccaaa aagaaattcg gacaagctta ttgctcgtaa aaaagactgg gatccaaaaa 4921 aatatggtgg ttttgatagt ccaacggtag cttattcagt cctagtggtt gctaaggtgg 4981 aaaaagggaa atcgaagaag ttaaaatccg ttaaagagtt actagggatc acaattatgg 5041 aaagaagttc ctttgaaaaa aatccgattg actttttaga agctaaagga tataaggaag 5101 ttaaaaaaga cttaatcatt aaactaccta aatatagtct ttttgagtta gaaaacggtc 5161 gtaaacggat gctggctagt gccggagaat tacaaaaagg aaatgagctg gctctgccaa 5221 gcaaatatgt gaatttttta tatttagcta gtcattatga aaagttgaag ggtagtccag 5281 aagataacga acaaaaacaa ttgtttgtgg agcagcataa gcattattta gatgagatta 5341 ttgagcaaat cagtgaattt tctaagcgtg ttattttagc agatgccaat ttagataaag 5401 ttcttagtgc atataacaaa catagagaca aaccaatacg tgaacaagca gaaaatatta 5461 ttcatttatt tacgttgacg aatcttggag ctcccgctgc ttttaaatat tttgatacaa 5521 caattgatcg taaacgatat acgtctacaa aagaagtttt agatgccact cttatccatc 5581 aatccatcac tggtctttat gaaacacgca ttgatttgag tcagctagga ggtgactgat 5641 ggctggttgg cgtactgttg tggtaaatac ccactcgaaa ttatcctata agaataatca 5701 tctgattttt aaggatgcct ataaaacgga gctgatccat ttatcagaaa ttgatatttt 5761 gttattagaa acgaccgata ttgtcttgtc cactatgctg gtaaaacggc tagtggatga 5821 gaatgtcctt gtcatattct gtgatgataa acgattacca acagctatgc tgatgccttt 5881 ttatggtcgt catgattcga gtttacagct tgggaaacaa atgtcctggt cagaaacagt 5941 caaatcgcag gtttggacga cgattattgc tcaaaagatt ttgaatcaat cttgctatct 6001 aggagcatgc tcctattttg aaaaatccca atctattatg gatttatatc atggtttgga 6061 aaattttgat ccgagtaatc gagaagggca tgcagcgaga atttatttta atacactttt 6121 tgggaacgat ttctcaagag atttggagca tccaatcaat gcaggtctgg attatggtta 6181 tactttatta ttgagtatgt ttgcgcgtga agtggttgtg tctggatgta tgactcagtt 6241 tgggcttaaa cacgctaatc agtttaatca gttcaatttt gctagcgata ttatggaacc 6301 atttaggcct ttagtggata agattgttta tgaaaatcga aatcagcctt ttcccaaaat 6361 aaagagagag ttatttactt tgttttcaga tacattttca tataatggta aagagatgta 6421 tctcacgaat attattagcg attatactaa aaaagttgtc aaagctctga ataatgaagg 6481 gaaaggagtt cctgaattta ggatatgagt tatagatata tgagaatgat acttatgttt 6541 gatatgccga cggacaccgc tgaggaacga aaagcctatc gaaaatttcg gaaattttta 6601 cttagtgaag ggtttatcat gcatcaattt tctatttata gtaagttgct gttgaataat 6661 acagctaaca atgccatgat tggtcggctg agggagcata atcctaataa aggaaatatt 6721 acattactaa cggtcacgga aaaacagttt gcacgaatga tttatttaca tggtgaaaga 6781 aataattgta ttgcaaactc cgatgaaaga cttgtatttc ttggggaggc ttttgatgaa 6841 tcttaatttt tccttactag atgaaccgat tccattaaga ggcggtacaa ttcttgtgct 6901 cgaagatgtc tgtgtatttt caaaaatagt gcaatattgt taccaatatg aggaagattc 6961 tgaacttaaa ttttttgatc acaagatgaa aacaatcaaa gaatcagaaa tcatgcttgt 7021 aacagatatt ttaggatttg atgttaactc ctcaaccatt ttaaaattga ttcatgcaga 7081 tttagaatct caatttaatg agaaacccga agtgaaatcg atgattgaca aattggttgc 7141 tacgattaca gaactgattg tctttgaatg cttagaaaat gaattagatt tagagtatga 7201 tgaaatcaca atcctggaat tgattaagtc cttaggagta aaagtagaaa cgcaaagtga 7261 tactattttt gaaaaatgtc ttgagatact tcaaattttc aaatatctca ctaagaaaaa 7321 gttgcttatt tttgtcaata gcggagcttt tctaacaaag gatgaagtgg ctagtttaca 7381 agagtatata tcattgacaa atttaacagt tctcttttta gaaccacgtg aactatatga 7441 ttttccgcag tatattttag atgaagatta tttcttaata actaaaaata tggtataata 7501 ctcttaataa atgcagtaat acaggggctt ttcaagactg aagtctagct gagacaaata 7561 gtgcgattac tagtagcggc cgctgcagtc cggcaaaaaa gggcaaggtg tcaccaccct 7621 gccctttttc tttaaaaccg aaaagattac ttcgcgttat gcaggcttcc tcgctcactg 7681 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 7741 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 7801 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccacagg ctccgccccc 7861 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 7921 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 7981 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 8041 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 8101 aaccccccgt tcagcccgac cgctgcgcct //