LOCUS       pTPH405c                7114 bp    DNA     circular     27-APR-2022
DEFINITION  [ColE1] sd8-psp-split-gIII CFN Npu 32aa intein dual stop NNNNCC 6,9
            Nme2 recoded N-term.
ACCESSION   urn.local...8x-7y8e687
VERSION     urn.local...8x-7y8e687
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     rep_origin      1..588
     source          1..>1302
     misc_feature    974..998
                     /note="Geneious type: primer_bind_reverse"
                     /standard_name="TPH1042"
     CDS             complement(1016..1207)
                     /note="Rop"
     primer_bind     1198..1217
                     /standard_name=1190
     source          <1303..>1317
     modified_base   1318..1332
                     /note="USER junction"
     source          <1318..7114
     regulatory      1333..1377
                     /note="Geneious type: terminator"
                     /note="rrnB1 transcriptional terminator"
     primer_bind     complement(1378..1406)
                     /standard_name="TPH1737"
     source          <1378..>1575
     misc_feature    1395..1406
                     /standard_name="USER Junction"
     primer_bind     1395..1418
                     /standard_name="TPH1736"
     misc_feature    1400..1575
                     /standard_name="insulated proD"
     primer_bind     1401..1425
                     /standard_name="TPH1625"
     primer_bind     complement(1461..1482)
                     /standard_name="AW040"
     regulatory      1462..1467
                     /note="Geneious type: promoter"
                     /standard_name="-35"
     misc_feature    1471..1482
                     /standard_name="USER"
     primer_bind     1471..1499
                     /standard_name="AW041"
     regulatory      1485..1490
                     /note="Geneious type: promoter"
                     /standard_name="proD -10"
     prim_transcript 1497
                     /standard_name="+1 TSS"
     primer_bind     complement(1559..1575)
                     /standard_name="TPH1729"
     misc_feature    1567..1575
                     /standard_name="USER Junction"
     primer_bind     complement(1574..1637)
                     /standard_name="TPH1985"
     misc_feature    1576..1589
                     /note="Geneious type: RBS"
                     /standard_name="sd8"
     source          <1576..>1589
     CDS             1590..>1658
                     /standard_name="III"
     source          <1590..>1658
     misc_feature    1620..1925
                     /note="Geneious type: motif"
                     /standard_name="NpuN"
     source          <1620..>1658
     source          join(<1620..>1658,<1620..>1658)
     source          join(<1620..>1658,<1620..>1658,<1620..>1658,<1620..>1658)
     gene            1630..>1658
                     /standard_name="Gblock-024"
     primer_bind     1630..1660
                     /standard_name="TPH1068"
     CDS             <1659..3380
                     /standard_name="III"
     source          <1659..>2135
     source          <1659..>1925
     source          join(<1659..>1925,<1659..>1925)
     source          join(<1659..>1925,<1659..>1925,<1659..>1925,<1659..>1925)
     source          <1926..>2021
     misc_feature    complement(<1945..1950)
                     /standard_name="Nme2 PAM"
     source          complement(<1948..>1950)
     source          join(complement(<1948..>1950),complement(<1948..>1950))
     source          <1948..>1950
     misc_feature    complement(1951..1973)
                     /standard_name="Nme2 Spacer"
     source          <1962..>1970
     misc_feature    1971..1982
                     /standard_name="USER Junction"
     primer_bind     1971..2000
                     /standard_name="TPH1871"
     misc_feature    2022..>2108
                     /note="Geneious type: motif"
                     /standard_name="npuC"
     source          <2022..>2126
     source          <2022..>2108
     source          join(<2022..>2108,<2022..>2108)
     primer_bind     2031..2051
                     /standard_name=1383
     primer_bind     complement(2089..2126)
                     /standard_name="TPH1080"
     misc_feature    <2109..2126
                     /note="Geneious type: motif"
                     /standard_name="npuC"
     source          <2109..>2126
     source          join(<2109..>2126,<2109..>2126)
     modified_base   2115..2126
     primer_bind     2115..2159
                     /standard_name="TPH1846"
     misc_feature    2127..2135
                     /standard_name="Scar"
     misc_feature    2149..2157
                     /standard_name="USER Junction"
     misc_feature    2164..2177
                     /standard_name="USER Junction"
     primer_bind     2164..2190
                     /standard_name="TPH1635"
     misc_feature    3248..3293
                     /note="modified to remove internal promoter"
     CDS             3380..5503
                     /note="xluxAB"
     sig_peptide     4463..4522
                     /note="GGGGS4 flexible linker"
     primer_bind     complement(5479..5516)
                     /standard_name="TPH1690"
     misc_feature    5504..5516
                     /note="USER linker"
                     /note="Geneious type: modified_base"
     primer_bind     5504..5527
                     /standard_name="TW0299"
     source          <5504..>5591
     source          <5504..>5516
     source          5517
     regulatory      5518..5551
                     /note="Geneious type: terminator"
                     /note="BBa_B1002"
     source          <5518..>5591
     regulatory      complement(5553..5591)
                     /note="Geneious type: terminator"
                     /note="BBa_B1006"
     modified_base   5592..5606
     primer_bind     5592..5611
                     /standard_name="TPH1509"
     source          <5592..>5759
     source          <5592..>5736
     source          <5592..>5608
     misc_feature    complement(5609..5736)
                     /note="Geneious type: misc_RNA"
                     /standard_name="Nme2Cas9 gRNA BB"
     source          <5609..>5615
     source          complement(<5609..>5615)
     source          complement(<5616..>5736)
     source          <5616..>5736
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     primer_bind     complement(5712..5731)
                     /standard_name="TPH1836"
     misc_feature    5720..5731
                     /standard_name="USER Junction"
     primer_bind     5720..5778
                     /standard_name="TPH1838"
     misc_feature    complement(5737..5759)
                     /standard_name="Nme2 Spacer"
     source          complement(<5737..>5759)
     source          <5737..>5759
     source          <5748..>5756
     misc_feature    complement(5760..5798)
                     /note="Geneious type: promoter prokaryotic"
                     /standard_name="lac promoter (no LacO)"
     regulatory      complement(5766..5771)
                     /note="Geneious type: -10_signal"
     regulatory      5790..5795
                     /note="Geneious type: -35_signal"
     misc_feature    complement(5814..5828)
                     /standard_name="CAP binding site"
     misc_feature    complement(5844..5936)
                     /standard_name="LacI fragment"
     misc_feature    5937..5996
                     /standard_name="Misc_Feature_13"
     misc_feature    5937..5953
                     /standard_name="USER linker"
     regulatory      complement(5954..5987)
                     /note="Geneious type: terminator"
                     /standard_name="BBa_B1002 terminator"
     misc_feature    5983..>6029
                     /standard_name="Misc_Feature_14"
     regulatory      complement(5988..6029)
                     /note="Geneious type: terminator"
                     /standard_name="tetA/orfL bidirectional terminator"
     primer_bind     complement(6021..6042)
                     /standard_name="TW1128"
     modified_base   6030..6042
                     /note="USER junction"
     primer_bind     6030..6065
                     /standard_name="TW128"
     source          <6030..>6042
     CDS             complement(6043..6903)
                     /note="bla (AmpR)"
                     /note="from pUC19"
     regulatory      complement(6904..7098)
                     /note="Geneious type: promoter"
                     /note="bla promoter"
                     /note="from pUC19"
     primer_bind     complement(7082..7114)
                     /standard_name="TPH1720"
     modified_base   7099..7114
                     /note="USER junction"
                     /note="Geneious type: misc_feature"
     primer_bind     7099..21
                     /standard_name="TPH1719"
     misc_feature    7100..7112
                     /note="SfiI"
                     /note="Keep unique in all constructs, can be used to
                     exchange antibiotic resistance markers or plasmid origins"
ORIGIN      
        1 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
       61 gcggtggttt gtttgccgga tcaagagcta ccacctcttt ttccgaaggt aactggcttc
      121 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
      181 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
      241 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
      301 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
      361 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
      421 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
      481 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
      541 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
      601 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
      661 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
      721 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
      781 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
      841 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
      901 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
      961 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     1021 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     1081 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     1141 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     1201 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     1261 aacgagagag gatgctcacg atacgggtta ctgatgatga acaagaggac atccggtaag
     1321 aggacatccg gtcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttgct
     1381 gaggagactt agggactctc acagctaaca ccacgtcgtc cctatctgct gccctaggtc
     1441 tatgagtggt tgctggataa ctttacgggc atgcataagg ctcgtataat atattcaggg
     1501 agaccacaac ggtttccctc tacaaataat tttgtttaac ttttactaga gtgggaccct
     1561 acctgcaggt gcagtaaagg aaaaaaaaaa tgaaaaaatt attatttgcg ataccgctgt
     1621 gcttaagcta cgaaactgaa atcttgaccg tcgaatatgg tctgctgcca atcggcaaga
     1681 ttgttgaaaa acgtattgaa tgtacggtct actcagtgga taacaacggc aatatctaca
     1741 cccagccggt ggcccagtgg catgaccgtg gtgaacagga agtgttcgaa tattgtctgg
     1801 aagacggatc tttaatccgt gccacaaagg atcacaaatt tatgactgta gatggtcaga
     1861 tgctcccaat cgacgaaatt tttgaacgcg aattagacct gatgcgcgtg gataatctcc
     1921 cgaattccgg aggatctagc ggggcaacct ctggctctga gtagtgagga acaagcgaga
     1981 gtgcaacacc tgaaagcagc gggggcagca gcggggggtc aatcaaaatt gccacgcgta
     2041 aatatttagg caaacagaat gtttatgata tcggtgtcga gcgcgatcat aatttcgcgc
     2101 tgaaaaacgg atttatagct agcaattgtt tcaacgttgt tcctttctat tctcactccg
     2161 ctgaaactgt tgaaagttgt ttagcaaaac cccatacaga aaattcattt actaacgtct
     2221 ggaaagacga caaaacttta gatcgttacg ctaactatga gggctgtctg tggaatgcta
     2281 caggcgttgt agtttgtact ggtgacgaaa ctcagtgtta cggtacatgg gttcctattg
     2341 ggcttgctat ccctgaaaat gagggtggtg gctctgaggg tggcggttct gagggtggcg
     2401 gttctgaggg tggcggtact aaacctcctg agtacggtga tacacctatt ccgggctata
     2461 cttatatcaa ccctctcgac ggcacttatc cgcctggtac tgagcaaaac cccgctaatc
     2521 ctaatccttc tcttgaggag tctcagcctc ttaatacttt catgtttcag aataataggt
     2581 tccgaaatag gcagggggca ttaactgttt atacgggcac tgttactcaa ggcactgacc
     2641 ccgttaaaac ttattaccag tacactcctg tatcatcaaa agccatgtat gacgcttact
     2701 ggaacggtaa attcagagac tgcgctttcc attctggctt taatgaggat ccattcgttt
     2761 gtgaatatca aggccaatcg tctgacctgc ctcaacctcc tgtcaatgct ggcggcggct
     2821 ctggtggtgg ttctggtggc ggctctgagg gtggtggctc tgagggtggc ggttctgagg
     2881 gtggcggctc tgagggaggc ggttccggtg gtggctctgg ttccggtgat tttgattatg
     2941 aaaagatggc aaacgctaat aagggggcta tgaccgaaaa tgccgatgaa aacgcgctac
     3001 agtctgacgc taaaggcaaa cttgattctg tcgctactga ttacggtgct gctatcgatg
     3061 gtttcattgg tgacgtttcc ggccttgcta atggtaatgg tgctactggt gattttgctg
     3121 gctctaattc ccaaatggct caagtcggtg acggtgataa ttcaccttta atgaataatt
     3181 tccgtcaata tttaccttcc ctccctcaat cggttgaatg tcgccctttt gtctttggcg
     3241 ctggtaaacc ttacgagttc agtatcgact gcgataagat caacctgttc cgcggtgtct
     3301 ttgcgtttct tttatatgtt gccaccttta tgtatgtatt ttctacgttt gctaacatac
     3361 tgcgtaataa ggagtcttaa tgaaatttgg aaactttttg cttacatacc aacctcccca
     3421 attttcccaa acagaggtaa tgaaacgttt ggttaaatta ggtcgcatct ctgaggagtg
     3481 tggttttgat accgtatggt tactggagca tcatttcacg gagtttggtt tgcttggtaa
     3541 cccttatgtc gctgctgcat atttacttgg cgcgactaaa aaattgaatg taggaactgc
     3601 cgctattgtt cttcccacag cccatccagt acgccaactt gaagatgtga atttattgga
     3661 tcaaatgtca aaaggacgat ttcggtttgg tatttgccga gggctttaca acaaggactt
     3721 tcgcgtattc ggcacagata tgaataacag tcgcgcctta gcggaatgct ggtacgggct
     3781 gataaagaat ggcatgacag agggatatat ggaagctgat aatgaacata tcaagttcca
     3841 taaggtaaaa gtaaaccccg cggcgtatag cagaggtggc gcaccggttt atgtggtggc
     3901 tgaatcagct tcgacgactg agtgggctgc tcaatttggc ctaccgatga tattaagttg
     3961 gattataaat actaacgaaa agaaagcaca acttgagctt tataatgaag tggctcaaga
     4021 atatgggcac gatattcata atatcgacca ttgcttatca tatataacat ctgtagatca
     4081 tgactcaatt aaagcgaaag agatttgccg gaaatttctg gggcattggt atgattctta
     4141 tgtgaatgct acgactattt ttgatgattc agaccaaaca agaggttatg atttcaataa
     4201 agggcagtgg cgtgactttg tattaaaagg acataaagat actaatcgcc gtattgatta
     4261 cagttacgaa atcaatcccg tgggaacgcc gcaggaatgt attgacataa ttcaaaaaga
     4321 cattgatgct acaggaatat caaatatttg ttgtggattt gaagctaatg gaacagtaga
     4381 cgaaattatt gcttccatga agctcttcca gtctgatgtc atgccatttc ttaaagaaaa
     4441 acaacgttcg ctattatatt atggcggtgg cggtagcggc ggtggcggta gcggcggtgg
     4501 cggtagcggc ggtggcggta gcaaatttgg attgttcttc cttaacttca tcaattcaac
     4561 aactgttcaa gaacagagta tagttcgcat gcaggaaata acggagtatg ttgataagtt
     4621 gaattttgaa cagattttag tgtatgaaaa tcatttttca gataatggtg ttgtcggcgc
     4681 tcctctgact gtttctggtt ttctgctcgg tttaacagag aaaattaaaa ttggttcatt
     4741 aaatcacatc attacaactc atcatcctgt ccgcatagcg gaggaagctt gcttattgga
     4801 tcagttaagt gaagggagat ttattttagg gtttagtgat tgcgaaaaaa aagatgaaat
     4861 gcattttttt aatcgcccgg ttgaatatca acagcaacta tttgaagagt gttatgaaat
     4921 cattaacgat gctttaacaa caggctattg taatccagat aacgattttt atagcttccc
     4981 taaaatatct gtaaatcccc atgcttatac gccaggcgga cctcggaaat atgtaacagc
     5041 aaccagtcat catattgttg agtgggcggc caaaaaaggt attcctctca tctttaagtg
     5101 ggatgattct aatgatgtta gatatgaata tgctgaaaga tataaagccg ttgcggataa
     5161 atatgacgtt gacctatcag agatagacca tcagttaatg atattagtta actataacga
     5221 agatagtaat aaagctaaac aagagacgcg tgcatttatt agtgattatg ttcttgaaat
     5281 gcaccctaat gaaaatttcg aaaataaact tgaagaaata attgcagaaa acgctgtcgg
     5341 aaattatacg gagtgtataa ctgcggctaa gttggcaatt gaaaagtgtg gtgcgaaaag
     5401 tgtattgctg tcctttgaac caatgaatga tttgatgagc caaaaaaatg taatcaatat
     5461 tgttgatgat aatattaaga agtaccacac ggaatatacc taaacggagc caatgtacgc
     5521 aaaaaacccc gcttcagcgg ggttttttcg ccaaaaaaaa ccccgccctg tcaggggcgg
     5581 ggtttttttt tagggcccta ctgactgtaa aaaaataaac gatgcccctt aaagcagaag
     5641 ctttaagggg cagagcgttg cggcacatct tttcagacgg ccttattgta gcaacggttc
     5701 tcatttcgtt tccgaaatga gaaagggagc tacaacctgg ctctgagtag tgaggaacac
     5761 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5821 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5881 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgcactt
     5941 aattaacggc actgcgaaaa aaccccgccg aagcggggtt ttttgcgaaa tataatgacc
     6001 ctcttgataa cccaagaggg cattttttaa tgcccatggc gtttaccaat gcttaatcag
     6061 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6121 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6181 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6241 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6301 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6361 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6421 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6481 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6541 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6601 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6661 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6721 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6781 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6841 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6901 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6961 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7021 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     7081 gcgtatcacg aggcccttag gccaaatagg ccgt
//
