LOCUS       pSLS.407                5197 bp    DNA     circular UNA 14-JUN-2019
DEFINITION  pRSFDuet-1_ec86(b3_v2)minusRT_cas1and2.
ACCESSION   urn.local...25-3x3fjhp
VERSION     urn.local...25-3x3fjhp
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     misc_RNA        28..109
                     /created_by="User"
                     /label="msdRNA"
     primer          complement(82..109)
                     /created_by="User"
                     /label="86_rev"
     primer          complement(88..>116)
                     /modified_by="User"
                     /label="msd_Rev"
     misc_feature    complement(99..155)
                     /created_by="Seth"
                     /label="msd"
     misc_feature    109..111
                     /Original_Bases="TGT"
                     /label="TGT"
                     /note="Geneious type: Editing History Replacement"
     motif           116..125
                     /created_by="User"
                     /label="Stem frag"
     motif           126..130
                     /created_by="User"
                     /label="loop"
     motif           complement(131..140)
                     /created_by="User"
                     /label="Stem frag"
     motif           complement(141..155)
                     /created_by="User"
                     /label="Flanking"
     primer          141..166
                     /created_by="User"
                     /label="86_fow"
     primer          <141..164
                     /modified_by="User"
                     /label="msd_Fow"
     misc_feature    157..168
                     /created_by="User"
                     /label="'primer'"
     gene            175..>187
                     /created_by="User"
                     /label="ec86RT"
     misc_feature    187^188
                     /Original_Bases="AATATTTGAACACTTTTAGATTGAGAAATCTCGGCCTACCT
                     GTCATGAACAATTTGCATGACATGTCTAAGGCGACTCGCATATCTGTTGAAACACTTC
                     GGTTGTTAATCTATACAGCTGATTTTCGCTATAGGATCTACACTGTAGAAAAGAAAGG
                     CCCAGAGAAGAGAATGAGAACCATTTACCAACCTTCTCGAGAACTTAAAGCCTTACAA
                     GGATGGGTTCTACGTAACATTTTAGATAAACTGTCGTCATCTCCTTTTTCTATTGGAT
                     TTGAAAAGCACCAATCTATTTTGAATAATGCTACCCCGCATATTGGGGCAAACTTTAT
                     ACTGAATATTGATTTGGAGGATTTTTTCCCAAGTTTAACTGCTAACAAAGTTTTTGGA
                     GTGTTCCATTCTCTTGGTTATAATCGACTAATATCTTCAGTTTTGACAAAAATATGTT
                     GTTATAAAAATCTGCTACCACAAGGTGCTCCATCATCACCTAAATTAGCTAATCTAAT
                     ATGTTCTAAACTTGATTATCGTATTCAGGGTTATGCAGGTAGTCGGGGCTTGATATAT
                     ACGAGATATGCCGATGATCTCACCTTATCTGCACAGTCTATGAAAAAGGTTGTTAAAG
                     CACGTGATTTTTTATTTTCTATAATCCCAAGTGAAGGATTGGTTATTAACTCAAAAAA
                     AACTTGTATTAGTGGGCCTCGTAGTCAGAGGAAAGTTACAGGTTTAGTTATTTCACAA
                     GAGAAAGTTGGGATAGGTAGAGAAAAATATAAAGAAATTAGAGCAAAGATACATCATA
                     TATTTTGCGGTAAGTCTTCTGAGATAGAACACGTTAGGGGATGGTTGTCATTTATTTT
                     AAGTGTGGATTCAAAAAGCCATAGGAGATTAATAACTTATATTAGCAAATTAGAAAAA
                     AAATATGGAAAGAACCCTTTAAATAAAGCGAAGACCTAATGACATGGGCAGCAGC"
                     /label="AATATTTGAACACTTTTAGATTGAGAAATCTCGGCCTACCTGTCATGAAC
                     AATTTGCATGACATGTCTAAGGCGACTCGCATATCTGTTGAAACACTTCGGTTGTTAA
                     TCTATACAGCTGATTTTCGCTATAGGATCTACACTGTAGAAAAGAAAGGCCCAGAGAA
                     GAGAATGAGAACCATTTACCAACCTTCTCGAGAACTTAAAGCCTTACAAGGATGGGTT
                     CTACGTAACATTTTAGATAAACTGTCGTCATCTCCTTTTTCTATTGGATTTGAAAAGC
                     ACCAATCTATTTTGAATAATGCTACCCCGCATATTGGGGCAAACTTTATACTGAATAT
                     TGATTTGGAGGATTTTTTCCCAAGTTTAACTGCTAACAAAGTTTTTGGAGTGTTCCAT
                     TCTCTTGGTTATAATCGACTAATATCTTCAGTTTTGACAAAAATATGTTGTTATAAAA
                     ATCTGCTACCACAAGGTGCTCCATCATCACCTAAATTAGCTAATCTAATATGTTCTAA
                     ACTTGATTATCGTATTCAGGGTTATGCAGGTAGTCGGGGCTTGATATATACGAGATAT
                     GCCGATGATCTCACCTTATCTGCACAGTCTATGAAAAAGGTTGTTAAAGCACGTGATT
                     TTTTATTTTCTATAATCCCAAGTGAAGGATTGGTTATTAACTCAAAAAAAACTTGTAT
                     TAGTGGGCCTCGTAGTCAGAGGAAAGTTACAGGTTTAGTTATTTCACAAGAGAAAGTT
                     GGGATAGGTAGAGAAAAATATAAAGAAATTAGAGCAAAGATACATCATATATTTTGCG
                     GTAAGTCTTCTGAGATAGAACACGTTAGGGGATGGTTGTCATTTATTTTAAGTGTGGA
                     TTCAAAAAGCCATAGGAGATTAATAACTTATATTAGCAAATTAGAAAAAAAATATGGA
                     AAGAACCCTTTAAATAAAGCGAAGACCTAATGACATGGGCAGCAGC"
                     /note="Geneious type: Editing History Deletion"
     misc_binding    188..205
                     /label="His-Tag"
     promoter        319..335
                     /label="T7\promoter"
     protein_bind    340..360
                     /label="lacO"
     RBS             391..396
     gene            404..1321
                     /label="Cas1"
     primer_bind     404..426
                     /label="Duet2_1263_fow"
     primer_bind     489..516
                     /label="Duet2_1178_fow"
     primer_bind     553..576
                     /label="Duet2_1114_fow"
     primer_bind     689..720
                     /label="Due2_978_fow"
     primer_bind     975..998
                     /label="Duet2_692_fow"
     primer_bind     1218..1242
                     /label="Duet2_449_fow"
     gene            1323..1607
                     /label="Cas2"
     primer_bind     1323..1353
                     /label="Duet2_344_fow"
     primer_bind     complement(1641..1666)
                     /label="Duet2_full_rev"
     CDS             1668..1796
                     /label="MCS-2\ORF"
     protein_bind    1734..1778
                     /label="S-Tag"
     primer          complement(1816..1834)
                     /label="T7\Terminator\primer"
     terminator      1830..1877
                     /label="T7\Terminator"
     CDS             complement(2107..2922)
                     /label="Kan"
     RBS             complement(2928..2932)
     -10_signal      complement(2964..2970)
                     /label="-10\Ap"
     -35_signal      complement(2987..2992)
                     /label="-35\Ap"
     rep_origin      3030..3779
                     /label="RSF\ori\-\RSF1030\Homology"
     misc_RNA        complement(3030..3728)
                     /label="RNA\II"
     -35_signal      3583..3588
                     /label="-35\RNA\I"
     -10_signal      3607..3612
                     /label="-10\RNA\I"
     misc_RNA        3619..3725
                     /label="RNA\I"
     CDS             complement(3975..5057)
                     /note="Wildtype lacI gene"
                     /label="lacI"
     primer_bind     5160..5197
                     /created_by="primer3"
                     /%GC="39.47"
                     /Tm="60.3"
                     /Hairpin="6.0"
                     /Primer-Dimer="4.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Sequence="GCGACTCCTGCATTAGGAAATTAATACGACTCACTATA"
                     /label="Primer"
     primer_bind     5164..5197
                     /created_by="primer3"
                     /%GC="35.29"
                     /Tm="56.63"
                     /Hairpin="6.0"
                     /Primer-Dimer="4.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Sequence="CTCCTGCATTAGGAAATTAATACGACTCACTATA"
                     /label="Primer"
     primer_bind     5168..5197
                     /created_by="primer3"
                     /%GC="30.0"
                     /Tm="52.98"
                     /Hairpin="6.0"
                     /Primer-Dimer="4.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Sequence="TGCATTAGGAAATTAATACGACTCACTATA"
                     /label="Primer"
     promoter        5181..5197
                     /label="T7\Promoter"
ORIGIN      
        1 gggcatttaa ctatgataag attccgtatg cgcaccctta gcgagaggtt tatcattaag
       61 gtcaacctct ggatgttgtt tcggcatcct gcattgaatc tgagttacaa gctgtttgtc
      121 gccagtcaga ctggcgacaa cccgtttttt ctgacgtaag ggtgcgcaac tttcatgaaa
      181 tccgctgcat caccatcatc accacagcca ggatccgaat tcgagctcgg cgcgcctgca
      241 ggtcgacaag cttgcggccg cataatgctt aagtcgaaca gaaagtaatc gtattgtaca
      301 cggccgcata atcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt
      361 ccccatctta gtatattagt taagtataag aaggagatat acaatgacct ggcttcccct
      421 taatcccatt ccactcaaag atcgcgtctc catgatcttt ctgcaatatg ggcagatcga
      481 tgtaatagat ggcgcgtttg tacttatcga caagacaggg atccgcactc atattcctgt
      541 tggctcggtt gcctgcatca tgctggaacc tggtacacgg gtttcgcatg cagctgtacg
      601 cctggctgcg caagttggaa cattgttggt atgggtgggg gaagcgggcg ttcgtgttta
      661 tgcttctggt cagcctggag gtgcgcgttc agataagctg ctctatcagg caaaacttgc
      721 tctggatgaa gatttgcgtc tgaaggtcgt acgtaaaatg tttgaacttc ggtttggaga
      781 acctgcgcct gcccggcgct ccgtagagca actcagaggt atagaaggca gtcgcgtgcg
      841 ggcaacctac gcacttctgg cgaagcaata cggcgtgaca tggaatggac gtcgctacga
      901 tccgaaagac tgggaaaagg gcgatacgat caaccaatgc attagcgctg caacttcctg
      961 tttatacggc gtaactgaag cggcgatact tgcagctggt tatgcaccag ctattgggtt
     1021 tgtgcataca ggaaagcctc tttcctttgt ttacgatatt gcagacatca ttaaatttga
     1081 cactgttgta ccgaaagctt ttgagatagc gcgtcgtaac cctggtgagc cggaccggga
     1141 agtccgtttg gcgtgcaggg atatttttcg cagtagtaaa acattagcca aattgattcc
     1201 gcttatagag gacgtgcttg ccgctggaga aatacaaccg ccggccccac ctgaagatgc
     1261 acagcctgtt gccattccgc ttcctgtttc actgggagat gcaggccatc ggagtagctg
     1321 aaatgagtat gttggtcgtg gtcactgaaa atgtacctcc gcgcttacga ggcagattag
     1381 ccatctggtt gttggaggta cgtgcagggg tatatgtagg tgatgtatcc gcaaaaattc
     1441 gtgaaatgat ctgggaacaa atagctggac tggcggaaga aggcaatgta gtgatggcat
     1501 gggcaacgaa tacggaaacg ggatttgagt tccagacatt tgggttaaac aggcgtaccc
     1561 cggtagattt ggatggttta aggttggtgt cttttttacc tgtttgagcg gccgcactcg
     1621 agtctggtaa agaaaccgct gctgcgaaat ttgaacgcca gcacattatg gcagatctca
     1681 attggatatc ggccggccac gcgatcgctg acgtcggtac cctcgagtct ggtaaagaaa
     1741 ccgctgctgc gaaatttgaa cgccagcaca tggactcgtc tactagcgca gcttaattaa
     1801 cctaggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     1861 tcttgagggg ttttttgctg aaacctcagg catttgagaa gcacacggtc acactgcttc
     1921 cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc
     1981 ctgaaccgac gacaagctga cgaccgggtc tccgcaagtg gcacttttcg gggaaatgtg
     2041 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat
     2101 taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
     2161 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     2221 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     2281 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     2341 gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac
     2401 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     2461 tgattgcgcc tgagcgagac gaaatacgcg gtcgctgtta aaaggacaat tacaaacagg
     2521 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     2581 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     2641 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     2701 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     2761 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     2821 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     2881 tcgcggccta gagcaagacg tttcccgttg aatatggctc atactcttcc tttttcaata
     2941 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3001 gaaaaataaa caaataggca tgcagcgctc ttccgcttcc tcgctcactg actcgctacg
     3061 ctcggtcgtt cgactgcggc gagcggtgtc agctcactca aaagcggtaa tacggttatc
     3121 cacagaatca ggggataaag ccggaaagaa catgtgagca aaaagcaaag caccggaaga
     3181 agccaacgcc gcaggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3241 gacgctcaag ccagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3301 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3361 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgttgg tatctcagtt
     3421 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3481 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3541 cactggcagc agccattggt aactgattta gaggactttg tcttgaagtt atgcacctgt
     3601 taaggctaaa ctgaaagaac agattttggt gagtgcggtc ctccaaccca cttaccttgg
     3661 ttcaaagagt tggtagctca gcgaaccttg agaaaaccac cgttggtagc ggtggttttt
     3721 ctttatttat gagatgatga atcaatcggt ctatcaagtc aacgaacagc tattccgtta
     3781 ctctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag ccccatacga
     3841 tataagttgt aattctcatg ttagtcatgc cccgcgccca ccggaaggag ctgactgggt
     3901 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     3961 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4021 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     4081 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     4141 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     4201 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc
     4261 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
     4321 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
     4381 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
     4441 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
     4501 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
     4561 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
     4621 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
     4681 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
     4741 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
     4801 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
     4861 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     4921 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     4981 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     5041 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     5101 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     5161 cgactcctgc attaggaaat taatacgact cactata
//
