LOCUS       Exported                6620 bp DNA     circular SYN 28-OCT-2020
DEFINITION  A new nucleotide sequence entered manually.
ACCESSION   urn.local...i-dfauj33
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6620)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jun 4, 2021 from SnapGene 5.3.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6620
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    29..335
                     /label=f1 origin
                     /note="Geneious type: origin of replication"
                     /note="/Imported_from=<a 
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla 
                     sMapper</a>"
                     /note="/Transferred_From=f1 origin"
                     /note="/Transferred_Similarity=99.67%"
     misc_feature    complement(560..1375)
                     /label=kan2 marker
                     /label=nonstandard type: marker
                     /note="/Imported_from=<a 
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla 
                     sMapper</a>"
                     /note="/Transferred_From=kan2 marker"
                     /note="/Transferred_Similarity=99.75%"
     misc_feature    1481..2100
                     /label=pBR322 origin
                     /note="Geneious type: origin of replication"
                     /note="/Imported_from=<a 
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla 
                     sMapper</a>"
                     /note="/Transferred_From=pBR322 origin"
                     /note="/Transferred_Similarity=99.68%"
     gene            complement(2515..2706)
                     /label=ROP
                     /note="/Imported_from=<a 
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla 
                     sMapper</a>"
                     /note="/Transferred_From=ROP"
                     /note="/Transferred_Similarity=100.00%"
     misc_feature    complement(3515..4606)
                     /label=lacI reg
                     /note="Geneious type: regulatory sequence"
                     /note="/Imported_from=<a 
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla 
                     sMapper</a>"
                     /note="/Transferred_From=lacI reg"
                     /note="/Transferred_Similarity=100.00%"
     promoter        4984..5002
                     /label=T7 prom
                     /note="/Imported_from=<a 
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla 
                     sMapper</a>"
                     /note="/Transferred_From=T7 prom"
                     /note="/Transferred_Similarity=100.00%"
     misc_feature    5002..5029
                     /label=lacO reg
                     /note="Geneious type: regulatory sequence"
                     /note="/Imported_from=<a 
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla 
                     sMapper</a>"
                     /note="/Transferred_From=lacO reg"
                     /note="/Transferred_Similarity=100.00%"
     gene            5047..5063
                     /label=T7_transl_en_RBS
                     /note="/Imported_from=<a 
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla 
                     sMapper</a>"
                     /note="/Transferred_From=T7_transl_en_RBS"
                     /note="/Transferred_Similarity=100.00%"
     misc_feature    5071..5073
                     /label=Start codon
                     /label=nonstandard type: marker
                     /note="/created_by=User"
     misc_feature    5083..5100
                     /label=6xHis tag
                     /label=nonstandard type: tag
                     /note="/Imported_from=<a 
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla 
                     sMapper</a>"
                     /note="/Transferred_From=6xHis tag"
                     /note="/Transferred_Similarity=94.44%"
     misc_feature    5109..5154
                     /label=J173
                     /label=nonstandard type: marker
                     /note="/created_by=User"
     misc_feature    5110..5130
                     /label=TEV cut site
                     /label=nonstandard type: marker
                     /note="/created_by=User"
     misc_feature    5131..6495
                     /label=CTGGTGCCGCGCGGCAGCCATATGGCTAGCATGACTGGTGGACAGCA
                     /label=CTGGTGCCGCGCGGCAGCCATATGGCTAGCATGACTGGTGGACAGCAAAT 
                     GGGTCGCGGATCCGAATTCGAGCTCCGTCGACAAGCTTGCGGCCGCACTCGAGCACCA 
                     CCACCACCACCACTGAGATCCGGCTGC
                     /note="Geneious type: Editing History Replacement"
                     /note="/Original_Bases=CTGGTGCCGCGCGGCAGCCATATGGCTAGCATGACT
                     GGTGG 
                     ACAGCAAATGGGTCGCGGATCCGAATTCGAGCTCCGTCGACAAGCTTGCGGCCGCACT 
                     CGAGCACCACCACCACCACCACTGAGATCCGGCTGC"
     misc_feature    5131..5445
                     /label=SUMO tag
                     /note="/created_by=User"
     misc_feature    5131..5154
                     /label=J173
                     /label=nonstandard type: marker
                     /note="/created_by=User"
     misc_feature    5446..6495
                     /label=PylB.3f2 (cloned from JH577 i.e. JH221)
                     /label=nonstandard type: marker
                     /note="/created_by=User"
                     /note="/modified_by=User"
     variation       5626..5628
                     /label=N61S
                     /label=nonstandard type: mutation
                     /note="/created_by=User"
     misc_feature    5626..5628
                     /label=Worse 61aa
                     /label=nonstandard type: marker
                     /note="/created_by=User"
                     /note="/modified_by=User"
     variation       5695..5697
                     /label=E84K
                     /label=nonstandard type: mutation
                     /note="/created_by=User"
     variation       5809..5811
                     /label=E122K
                     /label=nonstandard type: mutation
                     /note="/created_by=User"
     misc_feature    5830..5855
                     /label=617
                     /label=nonstandard type: motif
                     /note="/created_by=User"
     misc_feature    5968..5970
                     /label=E175K
                     /label=nonstandard type: motif
                     /note="/created_by=User"
     variation       5977..5979
                     /label=E178K
                     /label=nonstandard type: mutation
                     /note="/created_by=User"
                     /note="/modified_by=User"
     variation       6007..6009
                     /label=G188R
                     /label=nonstandard type: mutation
                     /note="/created_by=User"
                     /note="/modified_by=User"
     misc_feature    6007..6009
                     /label=188aa
                     /label=nonstandard type: marker
                     /note="/created_by=User"
                     /note="/modified_by=User"
     misc_feature    6141..6165
                     /label=382
                     /note="/created_by=User"
     variation       6196..6198
                     /label=E251D
                     /label=nonstandard type: mutation
                     /note="/created_by=User"
                     /note="/modified_by=User"
     misc_feature    complement(6469..6517)
                     /label=J174
                     /label=nonstandard type: marker
                     /note="/created_by=User"
     misc_feature    6496..6498
                     /label=Stop codon in pET28a ORF
                     /label=nonstandard type: marker
                     /note="/created_by=User"
                     /note="/modified_by=User"
     terminator      6553..6596
                     /label=T7 term
                     /note="/Imported_from=<a 
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla 
                     sMapper</a>"
                     /note="/Transferred_From=T7 term"
                     /note="/Transferred_Similarity=97.73%"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
      601 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
      661 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
      721 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
      781 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
      841 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
      901 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
      961 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     1021 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     1081 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     1141 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     1201 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     1261 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     1321 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
     1381 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
     1441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1621 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     1681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     2281 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta
     2341 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     2401 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     2461 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     2521 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     2581 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     2641 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     2701 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     2761 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     2821 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     2881 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     2941 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     3001 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     3061 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     3121 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     3181 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     3241 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     3301 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     3361 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     3421 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     3481 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3541 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3601 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     3661 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     3721 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     3781 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     3841 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     3901 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     3961 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     4021 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     4081 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     4141 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     4201 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     4261 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     4321 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     4381 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     4441 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     4501 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     4561 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     4621 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     4681 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     4741 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     4801 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     4861 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     4921 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
     4981 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
     5041 ttttgtttaa ctttaagaag gagatatacc atgggcagca gccatcatca tcatcatcac
     5101 agcagcggcg aaaacctgta ttttcagggc atgatgggca gcagcatgtc ggactcagaa
     5161 gtcaatcaag aagctaagcc agaggtcaag ccagaagtca agcctgagac tcacatcaat
     5221 ttaaaggtgt ccgatgggtc ttcagagatc ttcttcaaga tcaaaaagac cactccttta
     5281 agaaggctga tggaagcgtt cgctaaaaga cagggtaagg aaatggactc cttaagattc
     5341 ttgtacgacg gtattagaat tcaagctgat cagacccctg aagatttgga catggaggat
     5401 aacgatatta ttgaggctca cagagaacag attggtggtc atatgattaa gaagatggcg
     5461 actgaagatc tggatcgttt tggcgaaaaa attattgaag gatttcagtt atctgacgac
     5521 gatttacgcg ctttgcttag cctggaatcg gaagaagaat tggaaaagct gtattacgtc
     5581 gcacgtaaag tacgcaacca ttactttgga aatcgcgtat tcctgtcatg ttttatctac
     5641 ttcagcacat actgcaaaaa ccaatgtgcg ttttgctatt ataactgcaa aaacaagatt
     5701 aaccgctatc gtttgtcccg cgaagaaatc aaagagactt gcaaggccct taaaggtgcg
     5761 ggtttccata tgattgatct gactatgggg gaggatccct attattacaa ggaacctgat
     5821 cgcttcgtgg agctggtcca aatggttaag aatgagcttg gacttccaat catgatttcg
     5881 ccaggagtaa tggataatgc taccctttta aaagcacgcg aaaagggagc taatttcttc
     5941 gccttatatc aagaaaccta tgatcaagag ttgtacaaga agttgcgcgt aggtcagtcg
     6001 ttcaagaggc gctacaatac tcgtcgtttt gctaaagaac aagggtattg cattgaagac
     6061 ggtatcttaa cgggagtcgg gaacgatatc gagtcaacga tcaaatcttt acgcggcatg
     6121 aaatcaaatg atcccgacat ggttcgcgtg atgactttct taccccagga aggaactccc
     6181 cttgagcgtt ttaaggacaa tagtaacctg agcgaactga aaatcattgc gatcttgcgt
     6241 ctgatctttc cgaagtgctt aatccccgcg tcgttggacc tggaagggat cgacgggatg
     6301 gtacaccgtc ttaacgcagg tgctaatatc gtgactagca tccttcctag tgattcaaaa
     6361 cttgagggag ttgctaacta cgatcgtgac ctggaagagc gcgaccgcga cattaaaagc
     6421 gtcatcaagc gccttgaggg aatgggcatg gaaccagcgc cacaggcgga attcgagact
     6481 gtactgcgct gctaataaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     6541 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     6601 aggaggaact atatccggat
//
