LOCUS       Exported                9423 bp ds-DNA     circular SYN 28-OCT-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 9423)
  AUTHORS   Yvonne Nygard
  TITLE     Direct Submission
  JOURNAL   Exported Oct 28, 2022 from SnapGene 4.1.9
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9423
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     promoter        24..123
                     /label=tRNA Phe Promoter
     tRNA            124..214
                     /label=tRNA Phe
     misc_feature    215..280
                     /label=HDV Ribozyme
     misc_feature    285..304
                     /label=HO sgRNA
     misc_RNA        305..384
                     /label=sgRNA
     terminator      385..459
                     /label=tSNR52 terminator
                     /label=tSNR52
     misc_feature    460..484
                     /label=ConS scar
     promoter        489..1188
                     /label=ScPGK1 Promoter
     protein_bind    1194..1197
                     /label=BsaI
     CDS             1198..5298
                     /label=Cas9
     sig_peptide     5308..5358
                     /label=2x NLS Tag
     terminator      5374..5598
                     /label=ScPGK1 Terminator
     misc_feature    5603..5627
                     /label=ConE scar
     promoter        5651..6027
                     /label=AgTEF Promoter
                     /note="ApEinfo_fwdcolor=#B29990
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=#B29990"
     CDS             6030..6839
                     /label=KanR
                     /note="ApEinfo_fwdcolor=#E490FF
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=#E490FF"
     terminator      6840..7041
                     /label=AgTEF Terminator
                     /note="ApEinfo_fwdcolor=#B29990
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=#B29990"
     rep_origin      7046..7162
                     /label=CEN6
     rep_origin      7163..7553
                     /label=ARS4
     terminator      complement(7566..7662)
                     /label=AmpR Terminator
     CDS             complement(7663..8520)
                     /label=AmpR
     promoter        complement(8521..8651)
                     /label=AmpR Promoter
     rep_origin      complement(8652..9415)
                     /direction=LEFT
                     /label=ColE1
ORIGIN
        1 ccctgaattc gcatctagac tgaatgtgct tcagtattac attttttgcc ttcaacgcct
       61 tgattgttct atttttgcta ataataaatc tatttcatcg gactaaaagt ccattagttg
      121 taagcggatt tagctcagtt gggagagcgc cagactgaag aaaaacttcg gtcaagtcat
      181 ctggaggtcc tgtgttcgat ccacagaatt cgcagatggc cggcatggtc ccagcctcct
      241 cgctggcgcc ggctgggcaa caccttcggg tggcgaatgg gactgctcca gcattatagc
      301 atgcgtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa
      361 aagtggcacc gagtcggtgc tttttttatt ttttgtcact attgttatgt aaaatgccac
      421 ctctgacagt atggaacgca aacttctgtc tagtggatac tgaactggcc gataattgca
      481 gacgaacggt gagtaaggaa agagtgagga actatcgcat acctgcattt aaagatgccg
      541 atttgggcgc gaatccttta ttttggcttc accctcatac tattatcagg gccagaaaaa
      601 ggaagtgttt ccctccttct tgaattgatg ttaccctcat aaagcacgtg gcctcttatc
      661 gagaaagaaa ttaccgtcgc tcgtgatttg tttgcaaaaa gaacaaaact gaaaaaaccc
      721 agacacgctc gacttcctgt catcctattg attgcagctt ccaatttcgt cacacaacaa
      781 ggtcctagcg acggctcaca ggttttgtaa caagcaatcg aaggttctgg aatggcggga
      841 aagggtttag taccacatgc tatgatgccc actgtgatct ccagagcaaa gttcgttcga
      901 tcgtactgtt actctctctc tttcaaacag aattgtccga atcgtgtgac aacaacagcc
      961 tgttctcaca cactcttttc ttctaaccaa gggggtggtt tagtttagta gaacctcgtg
     1021 aaacttacat ttacatatat ataaacttgc ataaattggt caatgcaaga aatacatatt
     1081 tggtcttttc taattcgtag tttttcaagt tcttagatgc tttctttttc tcttttttac
     1141 agatcatcaa ggaagtaatt atctactttt tacaacaaat ataaaacaag atctatggac
     1201 aagaagtatt ctatcggact ggatatcggg actaatagcg tcgggtgggc cgtgatcact
     1261 gacgagtaca aggtgccctc taagaagttc aaggtgctcg ggaacaccga ccggcattcc
     1321 atcaagaaaa atctgatcgg agctctcctc tttgattcag gggaaaccgc tgaagcaacc
     1381 cgcctcaagc ggactgctag acggcggtac accaggagga agaaccggat ttgttacctt
     1441 caagagatat tctccaacga aatggcaaag gtcgacgaca gcttcttcca taggctggaa
     1501 gaatcattcc tcgtggaaga ggataagaag catgaacggc atcccatctt cggtaatatc
     1561 gtcgacgagg tggcctatca cgagaaatac ccaaccatct accatcttcg caaaaagctg
     1621 gtggactcaa ccgacaaggc agacctccgg cttatctacc tggccctggc ccacatgatc
     1681 aagttcagag gccacttcct gatcgagggc gacctcaatc ctgacaatag cgatgtggat
     1741 aaactgttca tccagctggt gcagacttac aaccagctct ttgaagagaa ccccatcaat
     1801 gcaagcggag tcgatgccaa ggccattctg tcagcccggc tgtcaaagag ccgcagactt
     1861 gagaatctta tcgctcagct gccgggtgaa aagaaaaatg gactgttcgg gaacctgatt
     1921 gctctttcac ttgggctgac tcccaatttc aagtctaatt tcgacctggc agaggatgcc
     1981 aagctgcaac tgtccaagga cacctatgat gacgatctcg acaacctcct ggcccagatc
     2041 ggtgaccaat acgccgacct tttccttgct gctaagaatc tttctgacgc catcctgctg
     2101 tctgacattc tccgcgtgaa cactgaaatc accaaggccc ctctttcagc ttcaatgatt
     2161 aagcggtatg atgagcacca ccaggacctg accctgctta aggcactcgt ccggcagcag
     2221 cttccggaga agtacaagga aatcttcttt gaccagtcaa agaatggata cgccggctac
     2281 atcgacggag gtgcctccca agaggaattt tataagttta tcaaacctat ccttgagaag
     2341 atggacggca ccgaagagct cctcgtgaaa ctgaatcggg aggatctgct gcggaagcag
     2401 cgcactttcg acaatgggag cattccccac cagatccatc ttggggagct tcacgccatc
     2461 cttcggcgcc aagaggactt ctaccccttt cttaaggaca acagggagaa gattgagaaa
     2521 attctcactt tccgcatccc ctactacgtg ggacccctcg ccagaggaaa tagccggttt
     2581 gcttggatga ccagaaagtc agaagaaact atcactccct ggaacttcga agaggtggtg
     2641 gacaagggag ccagcgctca gtcattcatc gaacggatga ctaacttcga taagaacctc
     2701 cccaatgaga aggtcctgcc gaaacattcc ctgctctacg agtactttac cgtgtacaac
     2761 gagctgacca aggtgaaata tgtcaccgaa gggatgagga agcccgcatt cctgtcaggc
     2821 gaacaaaaga aggcaattgt ggaccttctg ttcaagacca atagaaaggt gaccgtgaag
     2881 cagctgaagg aggactattt caagaaaatt gaatgcttcg actctgtgga gattagcggg
     2941 gtcgaagatc ggttcaacgc aagcctgggt acttaccatg atctgcttaa gatcatcaag
     3001 gacaaggatt ttctggacaa tgaggagaac gaggacatcc ttgaggacat tgtcctgact
     3061 ctcactctgt tcgaggaccg ggaaatgatc gaggagaggc ttaagaccta cgcccatctg
     3121 ttcgacgata aagtgatgaa gcaacttaaa cggagaagat ataccggatg gggacgcctt
     3181 agccgcaaac tcatcaacgg aatccgggac aaacagagcg gaaagaccat tcttgatttc
     3241 cttaagagcg acggattcgc taatcgcaac ttcatgcaac ttatccatga tgattccctg
     3301 acctttaagg aggacatcca gaaggcccaa gtgtctggac aaggtgactc actgcacgag
     3361 catatcgcaa atctggctgg ttcacccgct attaagaagg gtattctcca gaccgtgaaa
     3421 gtcgtggacg agctggtcaa ggtgatgggt cgccataaac cagagaacat tgtcatcgag
     3481 atggccaggg aaaaccagac tacccagaag ggacagaaga acagcaggga gcggatgaaa
     3541 agaattgagg aagggattaa ggagctcggg tcacagatcc ttaaagagca cccggtggaa
     3601 aacacccagc ttcagaatga gaagctctat ctgtactacc ttcaaaatgg acgcgatatg
     3661 tatgtggacc aagagcttga tatcaacagg ctctcagact acgacgtgga ccatatcgtc
     3721 cctcagagct tcctcaaaga cgactcaatt gacaataagg tgctgactcg ctcagacaag
     3781 aaccggggaa agtcagataa cgtgccctca gaggaagtcg tgaaaaagat gaagaactat
     3841 tggcgccagc ttctgaacgc aaagctgatc actcagcgga agttcgacaa tctcactaag
     3901 gctgagaggg gcggactgag cgaactggac aaagcaggat tcattaaacg gcaacttgtg
     3961 gagactcggc agattactaa acatgtcgcc caaatccttg actcacgcat gaataccaag
     4021 tacgacgaaa acgacaaact tatccgcgag gtgaaggtga ttaccctgaa gtccaagctg
     4081 gtcagcgatt tcagaaagga ctttcaattc tacaaagtgc gggagatcaa taactatcat
     4141 catgctcatg acgcatatct gaatgccgtg gtgggaaccg ccctgatcaa gaagtaccca
     4201 aagctggaaa gcgagttcgt gtacggagac tacaaggtct acgacgtgcg caagatgatt
     4261 gccaaatctg agcaggagat cggaaaggcc accgcaaagt acttcttcta cagcaacatc
     4321 atgaatttct tcaagaccga aatcaccctt gcaaacggtg agatccggaa gaggccgctc
     4381 atcgagacta atggggagac tggcgaaatc gtgtgggaca agggcagaga tttcgctacc
     4441 gtgcgcaaag tgctttctat gcctcaagtg aacatcgtga agaaaaccga ggtgcaaacc
     4501 ggaggctttt ctaaggaatc aatcctcccc aagcgcaact ccgacaagct cattgcaagg
     4561 aagaaggatt gggaccctaa gaagtacggc ggattcgatt caccaactgt ggcttattct
     4621 gtcctggtcg tggctaaggt ggaaaaagga aagtctaaga agctcaagag cgtgaaggaa
     4681 ctgctgggta tcaccattat ggagcgcagc tccttcgaga agaacccaat tgactttctc
     4741 gaagccaaag gttacaagga agtcaagaag gaccttatca tcaagctccc aaagtatagc
     4801 ctgttcgaac tggagaatgg gcggaagcgg atgctcgcct ccgctggcga acttcagaag
     4861 ggtaatgagc tggctctccc ctccaagtac gtgaatttcc tctaccttgc aagccattac
     4921 gagaagctga aggggagccc cgaggacaac gagcaaaagc aactgtttgt ggagcagcat
     4981 aagcattatc tggacgagat cattgagcag atttccgagt tttctaaacg cgtcattctc
     5041 gctgatgcca acctcgataa agtccttagc gcatacaata agcacagaga caaaccaatt
     5101 cgggagcagg ctgagaatat catccacctg ttcaccctca ccaatcttgg tgcccctgcc
     5161 gcattcaagt acttcgacac caccatcgac cggaaacgct atacctccac caaagaagtg
     5221 ctggacgcca ccctcatcca ccagagcatc accggacttt acgaaactcg gattgacctc
     5281 tcacagctcg gaggggatga gggagctgat ccaaaaaaga agagaaaggt agatccaaaa
     5341 aagaagagaa aggtataagg atcctaactc gagattgaat tgaattgaaa tcgatagatc
     5401 aatttttttc ttttctcttt ccccatcctt tacgctaaaa taatagttta ttttattttt
     5461 tgaatatttt ttatttatat acgtatatat agactattat ttatctttta atgattatta
     5521 agatttttat taaaaaaaaa ttcgctcctc ttttaatgcc tttatgcagt ttttttttcc
     5581 cattcgatat ttctatgtgc tgagcagtta cagagatgtt acgaaccact agtgcactgc
     5641 agtacagttt agcttgcctc gtccccgccg ggtcacccgg ccagcgacat ggaggcccag
     5701 aataccctcc ttgacagtct tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg
     5761 tacatttagc ccatacatcc ccatgtataa tcatttgcat ccatacattt tgatggccgc
     5821 acggcgcgaa gcaaaaatta cggctcctcg ctccagacct gcgagcaggg aaacgctccc
     5881 ctcacagacg cgttgaattg tccccacgcc gcgcccctgt agagaaatat aaaaggttag
     5941 gatttgccac tgaggttctt ctttcatata cttcctttta aaatcttgct aggatacagt
     6001 tctcacatca catccgaaca taaacaaaaa tgggtaagga aaagactcac gtttcgaggc
     6061 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
     6121 tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt
     6181 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
     6241 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     6301 atgcatggtt actcaccact gcgatccccg gcaaaacagc attccaggta ttagaagaat
     6361 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     6421 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctg gctcaggcgc
     6481 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     6541 ggcctgttga acaagtctgg aaagaaatgc ataagctttt gccattctca ccggattcag
     6601 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     6661 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     6721 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     6781 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaaa
     6841 gtaactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata gtttttttat
     6901 attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt tcgcctcgac
     6961 atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg
     7021 tgaatgctgg tcgctatact ggagtatcac gtgctataaa aataattata atttaaattt
     7081 tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag
     7141 tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta
     7201 tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaaa
     7261 accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta
     7321 tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt
     7381 tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac
     7441 tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta
     7501 ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtcccgagcg
     7561 gccgcgatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7621 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7681 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7741 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7801 ggacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7861 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7921 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7981 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8041 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8101 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8161 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8221 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8281 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8341 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8401 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8461 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8521 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8581 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8641 agtgccacct gtcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     8701 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     8761 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     8821 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     8881 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     8941 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     9001 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     9061 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     9121 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     9181 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     9241 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     9301 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     9361 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtgcggc
     9421 cgc
//
