LOCUS       pSLS.405                6192 bp    DNA     circular UNA 15-OCT-2019
DEFINITION  pRSFDuet-1_ec86(wt)_cas1and2.
ACCESSION   urn.local...25-3x3fjhp
VERSION     urn.local...25-3x3fjhp
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     primer_bind     4..37
                     /created_by="primer3"
                     /%GC="41.18"
                     /Tm="59.8"
                     /Hairpin="6.0"
                     /Primer-Dimer="4.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Sequence="CATTTAACTATGATAAGATTCCGTATGCGCACCC"
                     /label="Primer"
     misc_RNA        28..109
                     /created_by="User"
                     /label="msdRNA"
     primer          complement(29..40)
                     /created_by="User"
                     /label="primer1"
     primer          complement(82..109)
                     /created_by="User"
                     /label="86_rev"
     primer          complement(88..122)
                     /modified_by="User"
                     /label="msd_Rev"
     misc_feature    complement(99..184)
                     /created_by="User"
                     /label="msDNA"
     motif           99..115
                     /created_by="User"
                     /label="Flanking"
     motif           116..139
                     /created_by="User"
                     /modified_by="User"
                     /label="Stem"
     motif           140..145
                     /created_by="User"
                     /modified_by="User"
                     /label="Loop"
     motif           complement(146..169)
                     /created_by="User"
                     /modified_by="User"
                     /label="Stem"
     primer          163..193
                     /modified_by="User"
                     /label="msd_Fow"
     motif           complement(170..184)
                     /created_by="User"
                     /label="Flanking"
     primer          170..195
                     /created_by="User"
                     /label="86_fow"
     primer          186..197
                     /created_by="User"
                     /label="primer1"
     gene            204..1166
                     /created_by="User"
                     /label="ec86RT"
     primer_bind     complement(1134..1193)
                     /created_by="primer3"
                     /%GC="45.0"
                     /Tm="68.35"
                     /Hairpin="8.0"
                     /Primer-Dimer="4.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Sequence="GGAAAGAACCCTTTAAATAAAGCGAAGACCTAATGACATGGGCAGCA
                     GCCATCACCATCA"
                     /modified_by="User"
                     /label="Primer"
     primer_bind     complement(1134..1166)
                     /created_by="primer3"
                     /%GC="36.36"
                     /Tm="57.78"
                     /Hairpin="8.0"
                     /Primer-Dimer="1.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Sequence="GGAAAGAACCCTTTAAATAAAGCGAAGACCTAA"
                     /modified_by="User"
                     /label="Primer"
     primer_bind     1167..1188
                     /created_by="primer3"
                     /%GC="59.09"
                     /Tm="59.73"
                     /Hairpin="6.0"
                     /Primer-Dimer="4.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Sequence="TGACATGGGCAGCAGCCATCAC"
                     /label="Primer"
     misc_binding    1183..1200
                     /label="His-Tag"
     promoter        1314..1330
                     /label="T7\promoter"
     protein_bind    1335..1355
                     /label="lacO"
     RBS             1386..1391
     gene            1399..2316
                     /label="Cas1"
     primer_bind     1399..1421
                     /label="Duet2_1263_fow"
     primer_bind     1484..1511
                     /label="Duet2_1178_fow"
     primer_bind     1548..1571
                     /label="Duet2_1114_fow"
     primer_bind     1684..1715
                     /label="Due2_978_fow"
     primer_bind     1970..1993
                     /label="Duet2_692_fow"
     primer_bind     2213..2237
                     /label="Duet2_449_fow"
     gene            2318..2602
                     /label="Cas2"
     primer_bind     2318..2348
                     /label="Duet2_344_fow"
     primer_bind     complement(2636..2661)
                     /label="Duet2_full_rev"
     CDS             2663..2791
                     /label="MCS-2\ORF"
     protein_bind    2729..2773
                     /label="S-Tag"
     primer          complement(2811..2829)
                     /label="T7\Terminator\primer"
     terminator      2825..2872
                     /label="T7\Terminator"
     CDS             complement(3102..3917)
                     /label="Kan"
     RBS             complement(3923..3927)
     -10_signal      complement(3959..3965)
                     /label="-10\Ap"
     -35_signal      complement(3982..3987)
                     /label="-35\Ap"
     rep_origin      4025..4774
                     /label="RSF\ori\-\RSF1030\Homology"
     misc_RNA        complement(4025..4723)
                     /label="RNA\II"
     -35_signal      4578..4583
                     /label="-35\RNA\I"
     -10_signal      4602..4607
                     /label="-10\RNA\I"
     misc_RNA        4614..4720
                     /label="RNA\I"
     CDS             complement(4970..6052)
                     /note="Wildtype lacI gene"
                     /label="lacI"
     primer_bind     6155..6192
                     /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     6159..6192
                     /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     6163..6192
                     /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        6176..6192
                     /label="T7\Promoter"
ORIGIN      
        1 gggcatttaa ctatgataag attccgtatg cgcaccctta gcgagaggtt tatcattaag
       61 gtcaacctct ggatgttgtt tcggcatcct gcattgaatc tgagttactg tctgttttcc
      121 ttgttggaac ggagagcatc gcctgatgct ctccgagcca accaggaaac ccgttttttc
      181 tgacgtaagg gtgcgcaact ttcatgaaat ccgctgaata tttgaacact tttagattga
      241 gaaatctcgg cctacctgtc atgaacaatt tgcatgacat gtctaaggcg actcgcatat
      301 ctgttgaaac acttcggttg ttaatctata cagctgattt tcgctatagg atctacactg
      361 tagaaaagaa aggcccagag aagagaatga gaaccattta ccaaccttct cgagaactta
      421 aagccttaca aggatgggtt ctacgtaaca ttttagataa actgtcgtca tctccttttt
      481 ctattggatt tgaaaagcac caatctattt tgaataatgc taccccgcat attggggcaa
      541 actttatact gaatattgat ttggaggatt ttttcccaag tttaactgct aacaaagttt
      601 ttggagtgtt ccattctctt ggttataatc gactaatatc ttcagttttg acaaaaatat
      661 gttgttataa aaatctgcta ccacaaggtg ctccatcatc acctaaatta gctaatctaa
      721 tatgttctaa acttgattat cgtattcagg gttatgcagg tagtcggggc ttgatatata
      781 cgagatatgc cgatgatctc accttatctg cacagtctat gaaaaaggtt gttaaagcac
      841 gtgatttttt attttctata atcccaagtg aaggattggt tattaactca aaaaaaactt
      901 gtattagtgg gcctcgtagt cagaggaaag ttacaggttt agttatttca caagagaaag
      961 ttgggatagg tagagaaaaa tataaagaaa ttagagcaaa gatacatcat atattttgcg
     1021 gtaagtcttc tgagatagaa cacgttaggg gatggttgtc atttatttta agtgtggatt
     1081 caaaaagcca taggagatta ataacttata ttagcaaatt agaaaaaaaa tatggaaaga
     1141 accctttaaa taaagcgaag acctaatgac atgggcagca gccatcacca tcatcaccac
     1201 agccaggatc cgaattcgag ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa
     1261 tgcttaagtc gaacagaaag taatcgtatt gtacacggcc gcataatcga aattaatacg
     1321 actcactata ggggaattgt gagcggataa caattcccca tcttagtata ttagttaagt
     1381 ataagaagga gatatacaat gacctggctt ccccttaatc ccattccact caaagatcgc
     1441 gtctccatga tctttctgca atatgggcag atcgatgtaa tagatggcgc gtttgtactt
     1501 atcgacaaga cagggatccg cactcatatt cctgttggct cggttgcctg catcatgctg
     1561 gaacctggta cacgggtttc gcatgcagct gtacgcctgg ctgcgcaagt tggaacattg
     1621 ttggtatggg tgggggaagc gggcgttcgt gtttatgctt ctggtcagcc tggaggtgcg
     1681 cgttcagata agctgctcta tcaggcaaaa cttgctctgg atgaagattt gcgtctgaag
     1741 gtcgtacgta aaatgtttga acttcggttt ggagaacctg cgcctgcccg gcgctccgta
     1801 gagcaactca gaggtataga aggcagtcgc gtgcgggcaa cctacgcact tctggcgaag
     1861 caatacggcg tgacatggaa tggacgtcgc tacgatccga aagactggga aaagggcgat
     1921 acgatcaacc aatgcattag cgctgcaact tcctgtttat acggcgtaac tgaagcggcg
     1981 atacttgcag ctggttatgc accagctatt gggtttgtgc atacaggaaa gcctctttcc
     2041 tttgtttacg atattgcaga catcattaaa tttgacactg ttgtaccgaa agcttttgag
     2101 atagcgcgtc gtaaccctgg tgagccggac cgggaagtcc gtttggcgtg cagggatatt
     2161 tttcgcagta gtaaaacatt agccaaattg attccgctta tagaggacgt gcttgccgct
     2221 ggagaaatac aaccgccggc cccacctgaa gatgcacagc ctgttgccat tccgcttcct
     2281 gtttcactgg gagatgcagg ccatcggagt agctgaaatg agtatgttgg tcgtggtcac
     2341 tgaaaatgta cctccgcgct tacgaggcag attagccatc tggttgttgg aggtacgtgc
     2401 aggggtatat gtaggtgatg tatccgcaaa aattcgtgaa atgatctggg aacaaatagc
     2461 tggactggcg gaagaaggca atgtagtgat ggcatgggca acgaatacgg aaacgggatt
     2521 tgagttccag acatttgggt taaacaggcg taccccggta gatttggatg gtttaaggtt
     2581 ggtgtctttt ttacctgttt gagcggccgc actcgagtct ggtaaagaaa ccgctgctgc
     2641 gaaatttgaa cgccagcaca ttatggcaga tctcaattgg atatcggccg gccacgcgat
     2701 cgctgacgtc ggtaccctcg agtctggtaa agaaaccgct gctgcgaaat ttgaacgcca
     2761 gcacatggac tcgtctacta gcgcagctta attaacctag gctgctgcca ccgctgagca
     2821 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaacc
     2881 tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca
     2941 gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgacaa gctgacgacc
     3001 gggtctccgc aagtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3061 tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa ctcatcgagc
     3121 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     3181 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     3241 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     3301 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     3361 aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     3421 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     3481 acgcggtcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     3541 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     3601 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     3661 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     3721 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     3781 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     3841 tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc
     3901 cgttgaatat ggctcatact cttccttttt caatattatt gaagcattta tcagggttat
     3961 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggcatgcag
     4021 cgctcttccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcggcgagcg
     4081 gtgtcagctc actcaaaagc ggtaatacgg ttatccacag aatcagggga taaagccgga
     4141 aagaacatgt gagcaaaaag caaagcaccg gaagaagcca acgccgcagg cgtttttcca
     4201 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagccaga ggtggcgaaa
     4261 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4321 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4381 gctttctcat agctcacgct gttggtatct cagttcggtg taggtcgttc gctccaagct
     4441 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4501 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ttggtaactg
     4561 atttagagga ctttgtcttg aagttatgca cctgttaagg ctaaactgaa agaacagatt
     4621 ttggtgagtg cggtcctcca acccacttac cttggttcaa agagttggta gctcagcgaa
     4681 ccttgagaaa accaccgttg gtagcggtgg tttttcttta tttatgagat gatgaatcaa
     4741 tcggtctatc aagtcaacga acagctattc cgttactcta gatttcagtg caatttatct
     4801 cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc tcatgttagt
     4861 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
     4921 agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc
     4981 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5041 aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca
     5101 gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt
     5161 gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt
     5221 cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc ccggactcgg
     5281 taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa
     5341 cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc
     5401 cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca
     5461 gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac
     5521 ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac
     5581 tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag
     5641 cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc
     5701 gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca
     5761 tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt
     5821 gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc
     5881 ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca
     5941 ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct
     6001 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca
     6061 ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt
     6121 cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag gaaattaata
     6181 cgactcacta ta
//
