LOCUS       pSBK.016                5212 bp    DNA     circular UNA 29-OCT-2019
DEFINITION  pRSFDuet-1_ec86(b3_v35lp)minusRT_cas1and2 (barcode GCA-TGC).
ACCESSION   urn.local...25-3x3fjhp
VERSION     urn.local...25-3x3fjhp
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     misc_signal     1..27
                     /created_by="User"
                     /Transferred_From="template"
                     /Transferred_Similarity="100.00%"
                     /label="T7 to msr"
     misc_rna        28..109
                     /created_by="User"
                     /Transferred_From="template"
                     /Transferred_Similarity="100.00%"
                     /label="msdRNA"
     misc_feature    29..40
                     /created_by="User"
                     /Transferred_From="template"
                     /Transferred_Similarity="100.00%"
                     /label="'primer'"
     primer          complement(82..>98)
                     /created_by="User"
                     /label="86_rev"
     primer          complement(88..>98)
                     /modified_by="User"
                     /label="msd_Rev"
     misc_feature    complement(99..155)
                     /created_by="User"
                     /modified_by="Seth"
                     /label="msd"
     misc_feature    109..139
                     /Original_Bases="TGTCTGTTTGTCGCCAGTCAGACTGGCGACA"
                     /label="TGTCTGTTTGTCGCCAGTCAGACTGGCGACA"
                     /note="Geneious type: Editing History Replacement"
     primer          <156..166
                     /created_by="User"
                     /label="86_fow"
     primer          <156..164
                     /modified_by="User"
                     /label="msd_Fow"
     misc_feature    157..183
                     /created_by="User"
                     /label="'primer'"
     misc_feature    169..183
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     gene            190..>202
                     /created_by="User"
                     /label="ec86RT"
     misc_feature    202^203
                     /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    203..220
                     /label="His-Tag"
     promoter        334..350
                     /label="T7\promoter"
     protein_bind    355..375
                     /label="lacO"
     RBS             406..411
     gene            419..1336
                     /label="Cas1"
     primer_bind     419..441
                     /label="Duet2_1263_fow"
     primer_bind     504..531
                     /label="Duet2_1178_fow"
     primer_bind     568..591
                     /label="Duet2_1114_fow"
     primer_bind     704..735
                     /label="Due2_978_fow"
     primer_bind     990..1013
                     /label="Duet2_692_fow"
     primer_bind     1233..1257
                     /label="Duet2_449_fow"
     gene            1338..1622
                     /label="Cas2"
     primer_bind     1338..1368
                     /label="Duet2_344_fow"
     primer_bind     complement(1656..1681)
                     /label="Duet2_full_rev"
     CDS             1683..1811
                     /label="MCS-2\ORF"
     protein_bind    1749..1793
                     /label="S-Tag"
     primer          complement(1831..1849)
                     /label="T7\Terminator\primer"
     terminator      1845..1892
                     /label="T7\Terminator"
     CDS             complement(2122..2937)
                     /label="Kan"
     RBS             complement(2943..2947)
     -10_signal      complement(2979..2985)
                     /label="-10\Ap"
     -35_signal      complement(3002..3007)
                     /label="-35\Ap"
     rep_origin      3045..3794
                     /label="RSF\ori\-\RSF1030\Homology"
     misc_RNA        complement(3045..3743)
                     /label="RNA\II"
     -35_signal      3598..3603
                     /label="-35\RNA\I"
     -10_signal      3622..3627
                     /label="-10\RNA\I"
     misc_RNA        3634..3740
                     /label="RNA\I"
     CDS             complement(3990..5072)
                     /note="Wildtype lacI gene"
                     /label="lacI"
     cds             complement(<4986..5072)
                     /note="Wildtype lacI gene"
                     /Transferred_From="template"
                     /Transferred_Similarity="100.00%"
                     /label="lacI"
     cds             complement(5022..5072)
                     /created_by="User"
                     /modified_by="User"
                     /Transferred_From="template"
                     /Transferred_Similarity="100.00%"
                     /label="end of lacI"
     misc_signal     5073..5195
                     /created_by="User"
                     /Transferred_From="template"
                     /Transferred_Similarity="100.00%"
                     /label="lacI to T7"
     primer_bind     5175..5212
                     /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     5179..5212
                     /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     5183..5212
                     /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        5196..5212
                     /label="T7\Promoter"
ORIGIN      
        1 gggcatttaa ctatgataag attccgtatg cgcaccctta gcgagaggtt tatcattaag
       61 gtcaacctct ggatgttgtt tcggcatcct gcattgaatc tgagttacaa gaaaatggag
      121 aggtgcaatg cacctctcca cccgtttttt ctgacgtaag ggtgcgcata cggaatctta
      181 tcaactttca tgaaatccgc tgcatcacca tcatcaccac agccaggatc cgaattcgag
      241 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag
      301 taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt
      361 gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga gatatacaat
      421 gacctggctt ccccttaatc ccattccact caaagatcgc gtctccatga tctttctgca
      481 atatgggcag atcgatgtaa tagatggcgc gtttgtactt atcgacaaga cagggatccg
      541 cactcatatt cctgttggct cggttgcctg catcatgctg gaacctggta cacgggtttc
      601 gcatgcagct gtacgcctgg ctgcgcaagt tggaacattg ttggtatggg tgggggaagc
      661 gggcgttcgt gtttatgctt ctggtcagcc tggaggtgcg cgttcagata agctgctcta
      721 tcaggcaaaa cttgctctgg atgaagattt gcgtctgaag gtcgtacgta aaatgtttga
      781 acttcggttt ggagaacctg cgcctgcccg gcgctccgta gagcaactca gaggtataga
      841 aggcagtcgc gtgcgggcaa cctacgcact tctggcgaag caatacggcg tgacatggaa
      901 tggacgtcgc tacgatccga aagactggga aaagggcgat acgatcaacc aatgcattag
      961 cgctgcaact tcctgtttat acggcgtaac tgaagcggcg atacttgcag ctggttatgc
     1021 accagctatt gggtttgtgc atacaggaaa gcctctttcc tttgtttacg atattgcaga
     1081 catcattaaa tttgacactg ttgtaccgaa agcttttgag atagcgcgtc gtaaccctgg
     1141 tgagccggac cgggaagtcc gtttggcgtg cagggatatt tttcgcagta gtaaaacatt
     1201 agccaaattg attccgctta tagaggacgt gcttgccgct ggagaaatac aaccgccggc
     1261 cccacctgaa gatgcacagc ctgttgccat tccgcttcct gtttcactgg gagatgcagg
     1321 ccatcggagt agctgaaatg agtatgttgg tcgtggtcac tgaaaatgta cctccgcgct
     1381 tacgaggcag attagccatc tggttgttgg aggtacgtgc aggggtatat gtaggtgatg
     1441 tatccgcaaa aattcgtgaa atgatctggg aacaaatagc tggactggcg gaagaaggca
     1501 atgtagtgat ggcatgggca acgaatacgg aaacgggatt tgagttccag acatttgggt
     1561 taaacaggcg taccccggta gatttggatg gtttaaggtt ggtgtctttt ttacctgttt
     1621 gagcggccgc actcgagtct ggtaaagaaa ccgctgctgc gaaatttgaa cgccagcaca
     1681 ttatggcaga tctcaattgg atatcggccg gccacgcgat cgctgacgtc ggtaccctcg
     1741 agtctggtaa agaaaccgct gctgcgaaat ttgaacgcca gcacatggac tcgtctacta
     1801 gcgcagctta attaacctag gctgctgcca ccgctgagca ataactagca taaccccttg
     1861 gggcctctaa acgggtcttg aggggttttt tgctgaaacc tcaggcattt gagaagcaca
     1921 cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta
     1981 tttaacgacc ctgccctgaa ccgacgacaa gctgacgacc gggtctccgc aagtggcact
     2041 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2101 tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     2161 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     2221 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     2281 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     2341 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     2401 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     2461 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcggtcgc tgttaaaagg
     2521 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     2581 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     2641 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     2701 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     2761 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     2821 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     2881 catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcatact
     2941 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3001 atttgaatgt atttagaaaa ataaacaaat aggcatgcag cgctcttccg cttcctcgct
     3061 cactgactcg ctacgctcgg tcgttcgact gcggcgagcg gtgtcagctc actcaaaagc
     3121 ggtaatacgg ttatccacag aatcagggga taaagccgga aagaacatgt gagcaaaaag
     3181 caaagcaccg gaagaagcca acgccgcagg cgtttttcca taggctccgc ccccctgacg
     3241 agcatcacaa aaatcgacgc tcaagccaga ggtggcgaaa cccgacagga ctataaagat
     3301 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3361 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3421 gttggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3481 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3541 gacacgactt atcgccactg gcagcagcca ttggtaactg atttagagga ctttgtcttg
     3601 aagttatgca cctgttaagg ctaaactgaa agaacagatt ttggtgagtg cggtcctcca
     3661 acccacttac cttggttcaa agagttggta gctcagcgaa ccttgagaaa accaccgttg
     3721 gtagcggtgg tttttcttta tttatgagat gatgaatcaa tcggtctatc aagtcaacga
     3781 acagctattc cgttactcta gatttcagtg caatttatct cttcaaatgt agcacctgaa
     3841 gtcagcccca tacgatataa gttgtaattc tcatgttagt catgccccgc gcccaccgga
     3901 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     3961 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4021 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4081 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     4141 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     4201 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     4261 actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     4321 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     4381 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     4441 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     4501 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     4561 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     4621 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     4681 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     4741 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     4801 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     4861 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     4921 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     4981 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     5041 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     5101 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     5161 acgctctccc ttatgcgact cctgcattag gaaattaata cgactcacta ta
//
