LOCUS       Exported                9156 bp ds-DNA     circular SYN 26-JUL-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9156)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Dec 16, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Annotated with pLannotate v1.2.0
FEATURES             Location/Qualifiers
     source          1..9156
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      complement(350..547)
                     /label=TEF terminator
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=terminator"
     CDS             complement(554..1118)
                     /codon_start=1
                     /label=NrsR (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=99.8"
                     /note="/match_length=99.1"
                     /note="/other=CDS"
                     /translation="TTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGFTL
                     REVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNRRL
                     TVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFTLC
                     GLDTALYDGTASDGEQALYMSMPCP"
     promoter        complement(1125..1468)
                     /label=TEF promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     CDS             1511..1577
                     /codon_start=1
                     /label=lacZ-alpha (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=38.5"
                     /note="/other=CDS"
                     /translation="LA**RRGPHRSPFPTVAQPEWR"
     rep_origin      1581..2009
                     /label=f1 ori
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
     CDS             complement(2167..2970)
                     /codon_start=1
                     /label=URA3
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.8"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(2971..3186)
                     /label=URA3 promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     misc_feature    complement(3448..3951)
                     /label=CEN/ARS
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=misc_feature"
     promoter        3988..4092
                     /label=AmpR promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     CDS             4093..4953
                     /codon_start=1
                     /label=AmpR
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.8"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5124..5712
                     /direction=RIGHT
                     /label=ori
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.8"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
     misc_feature    5772..7792
                     /label=ACT1pr-yZ3EV-3'UTR
                     /note="Yeast codon-optimized Z3EV transcription factorunder
                     the control of the ACT1 promoter; has the A. gossypii TEF3 
                     3'UTR"
     misc_feature    7799..8497
                     /label=Z3 promoter
     misc_feature    8542..8639
                     /label=Retron Ec86
     misc_feature    8655..8755
                     /label=ADE2 repair template (introduces frameshift)
     misc_feature    8705
                     /label=Frameshift insertion
                     /note="""C"" inserted to introduce frameshift, located in 
                     the PAM seq."
     misc_feature    8756..8789
                     /label=MSD 3'
     misc_feature    8790..8809
                     
                     /note="Targets cas9 to cut at nt 450 of ADE2 gene."
     misc_feature    8810..8971
                     /label=sgRNA and HDV
                     /note="structural guide RNA and HDV ribozyme"
     misc_feature    join(8972..9156,1..170)
                     /label=tGAL7
ORIGIN
        1 aaaaaaaaaa aaagaaaaaa gaattttagg aatacaatgc agcttgtaag taaaatctgg
       61 aatattcata tcgccacaac ttcttatgct tataaaagca ctaatgcctg aatttatgtt
      121 gaaaatatgt gtcacaaata aagaaactgt gacatctgac acatttccac gtacccaatt
      181 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
      241 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagtc
      301 gagagctcgt tttcgacact ggatggcggc gttagtatcg aatcgacagc agtatagcga
      361 ccagcattca catacgattg acgcatgata ttactttctg cgcacttaac ttcgcatctg
      421 ggcagatgat gtcgaggcga aaaaaaatat aaatcacgct aacatttgat taaaatagaa
      481 caactacaat ataaaaaaac tatacaaatg acaagttctt gaaaacaaga atctttttat
      541 tgtcagtact gataggggca gggcatgctc atgtagagcg cctgctcgcc gtccgaggcg
      601 gtgccgtcgt acagggcggt gtccaggccg cagagggtga accccatccg ccggtacgcg
      661 tggatcgccg gtgcgttgac gttggtgacc tccagccaga ggtgcccggc gccccgctcg
      721 cgggcgaact ccgtcgcgag ccccatcaac gcgcgcccga ccccgtgccc ccggtgctcc
      781 ggggcgacct cgatgtcctc gacggtcagc cggcggttcc agccggagta cgagacgacc
      841 acgaagcccg ccaggtcgcc gtcgtccccg tacgcgacga acgtccggga gtccgggtcg
      901 ccgtcctccc cgtcgtccga ttcgtcgtcc gattcgtcgt cggggaacac cttggtcagg
      961 ggcgggtcca ccggcacctc ccgcagggtg aagccgtccc cggtggcggt gacgcggaag
     1021 acggtgtcgg tggtgaagga cccatccagt gcctcgatgg cctcggcgtc ccccgggaca
     1081 ctggtgcggt accggtaagc cgtgtcgtca agagtggtac ccatggttgt ttatgttcgg
     1141 atgtgatgtg agaactgtat cctagcaaga ttttaaaagg aagtatatga aagaagaacc
     1201 tcagtggcaa atcctaacct tttatatttc tctacagggg cgcggcgtgg ggacaattca
     1261 acgcgtctgt gaggggagcg tttccctgct cgcaggtctg cagcgaggag ccgtaatttt
     1321 tgcttcgcgc cgtgcggcca tcaaaatgta tggatgcaaa tgattataca tggggatgta
     1381 tgggctaaat gtacgggcga cagtcacatc atgcccctga gctgcgcacg tcaagactgt
     1441 caaggagggt attctgggcc tccatgtcgc tggccgggtg acccggcggg gacgaggcaa
     1501 gctaaacaga ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     1561 gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     1621 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     1681 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     1741 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     1801 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     1861 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     1921 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     1981 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg
     2041 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatagg gtaataactg
     2101 atataattaa attgaagctc taatttgtga gtttagtata catgcattta cttataatac
     2161 agttttttag ttttgctggc cgcatcttct caaatatgct tcccagcctg cttttctgta
     2221 acgttcaccc tctaccttag catcccttcc ctttgcaaat agtcctcttc caacaataat
     2281 aatgtcagat cctgtagaga ccacatcatc cacggttcta tactgttgac ccaatgcgtc
     2341 tcccttgtca tctaaaccca caccgggtgt cataatcaac caatcgtaac cttcatctct
     2401 tccacccatg tctctttgag caataaagcc gataacaaaa tctttgtcgc tcttcgcaat
     2461 gtcaacagta cccttagtat attctccagt agatagggag cccttgcatg acaattctgc
     2521 taacatcaaa aggcctctag gttcctttgt tacttcttct gccgcctgct tcaaaccgct
     2581 aacaatacct gggcccacca caccgtgtgc attcgtaatg tctgcccatt ctgctattct
     2641 gtatacaccc gcagagtact gcaatttgac tgtattacca atgtcagcaa attttctgtc
     2701 ttcgaagagt aaaaaattgt acttggcgga taatgccttt agcggcttaa ctgtgccctc
     2761 catggaaaaa tcagtcaaga tatccacatg tgtttttagt aaacaaattt tgggacctaa
     2821 tgcttcaact aactccagta attccttggt ggtacgaaca tccaatgaag cacacaagtt
     2881 tgtttgcttt tcgtgcatga tattaaatag cttggcagca acaggactag gatgagtagc
     2941 agcacgttcc ttatatgtag ctttcgacat gatttatctt cgtttcctgc aggtttttgt
     3001 tctgtgcagt tgggttaaga atactgggca atttcatgtt tcttcaacac tacatatgcg
     3061 tatatatacc aatctaagtc tgtgctcctt ccttcgttct tccttctgtt cggagattac
     3121 cgaatcaaaa aaatttcaag gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga
     3181 attgaaaagg tggtatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3241 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     3301 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     3361 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     3421 gtcatgataa taatggtttc ttagacggat cgcttgcctg taacttacac gcgcctcgta
     3481 tcttttaatg atggaataat ttgggaattt actctgtgtt tatttatttt tatgttttgt
     3541 atttggattt tagaaagtaa ataaagaagg tagaagagtt acggaatgaa gaaaaaaaaa
     3601 taaacaaagg tttaaaaaat ttcaacaaaa agcgtacttt acatatatat ttattagaca
     3661 agaaaagcag attaaataga tatacattcg attaacgata agtaaaatgt aaaatcacag
     3721 gattttcgtg tgtggtcttc tacacagaca agatgaaaca attcggcatt aatacctgag
     3781 agcaggaaga gcaagataaa aggtagtatt tgttggcgat ccccctagag tcttttacat
     3841 cttcggaaaa caaaaactat tttttcttta atttcttttt ttactttcta tttttaattt
     3901 atatatttat attaaaaaat ttaaattata attattttta tagcacgtga tgaaaaggac
     3961 ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4021 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4081 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4141 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4201 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4261 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4321 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4381 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4441 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4501 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4561 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4621 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4681 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4741 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4801 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4861 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4921 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4981 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5041 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5101 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5161 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5221 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5281 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5341 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5401 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5461 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5521 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5581 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5641 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5701 agcctatgga aaaaggccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5761 tttgctcgct cgcgtcgatc tcccctcaag tggcaaactg cgactgatat gaaagtaaag
     5821 tatcggacct tctattacgg aatgtataca aacgctacag gatacagtga tattaaatag
     5881 aaaacaagtc tacaattaaa tggcagaaag aagaaccagc ttactactaa ccaccatact
     5941 catcaatacc cagagcatcg gtgaacatct gctcaaattc gaagtcagcc atatccagcg
     6001 caccgtaagg tgctgaatca tgtggagtga agccaggccc tggagagtcc ccgtcaccca
     6061 acatatctaa atcaaaatcg tctaaagcgt cagcgtgggc catggcaacg tcttcaccgt
     6121 ccaaatgcag ttcgctagat gtagaccctg ctgtcgctag gtgggattgg tcggtttctt
     6181 caacggaggc tccgcctcta gaagttggtg catgcaacct gtgagcgtcc agcatttcta
     6241 gtaataaatc atacagagga acgacgtttt tacacttcat agaatacagg tgctccatac
     6301 ccttgttaga catatgtcta atgtgtgaca aaattagtag taactgagct agcctctgat
     6361 gctgttgctg cagagtcaat cctgctttcg ccataaggtg aatcaatgta tcagtgatct
     6421 tgtccaacac tctatgtata tggtctttct cttccaggct tttcagagtt gagcttaaaa
     6481 atgtatagac accgctattt aacaaaataa ttgatttcag gcaaacgaat tcttctcctt
     6541 gcaaattcat cattctgaac ctggagctag tggcaagaag catatcaaaa atctcaacca
     6601 taccttctac gcatttacct tggttcctgt ccaataataa attaggtgca aataataatt
     6661 ttacagggtg ctccatactt ctccacacta atccgatcat caatatttct aaccacgcgc
     6721 attcaagaag atgtacctgg tcatgtaatg tcaagtcaac aaagccggga actcttttgg
     6781 cccaattaat catatgaact aattccctgt ctgctagatt tgtcaaaaga cccatcatgg
     6841 aagcctcgga aaaaggacga gttgggtcgt attcactgta caatataggt ggttcagcat
     6901 caagtagtgc ggaaaccatt tgatcagcgg tcaaagacag tgcaagagaa ttcttcttag
     6961 atcttttgat cattaatggg cttggccaca agttggctgc cctcatgtct ccagcgcttg
     7021 gatcttctag ggttgaggct gctgccggag tgcccccacc tcctgtatgg atctttgtat
     7081 gtctctttct ttcatcagat ctggcgaact ttcttccaca aatgtcacac gcgaacggct
     7141 tctcgccggt gtgggttcta atgtgcgtag tcaaatggtc acttctgctg aagtttctca
     7201 tacagattct acattggaaa ggcttttggc cggtgtgaat acgaatatgc ctagtcagct
     7261 cgtcagatct gctgaatctc ctatcacaac tttctacagg gcaagcgtag ggtctcgttc
     7321 ccattgttaa ttcagtaaat tttcgatctt gggaagaaaa aagcagtaag cgtgaaaaat
     7381 ctaaaagctg atgtagtaga agatcctatt ctttaacaaa gattgacctt ttctttttct
     7441 tcttggtttg agtagaaagg ggaaggaaga atacaagaga gaggaaaaaa aggaagataa
     7501 aaagagagcg tgatataaat gaatatatat taaacaagag agattgggaa ggaaaggatc
     7561 aaacaaaccc aaaaatattt caaaaaggag agagagaggc gagtttggtt tcaaaacggt
     7621 ttatttattt atgcaagagg acgtggaaga aaaagaagaa ggaagaaaaa aatttgaaag
     7681 aaaaaaacgc gtggcgggta aagaagaaaa tggaaaatag aggccgggtg acagagaaat
     7741 attgagggtt aattggaaaa tatgttaggg tgaggcatat gtttttaaag ggactagttt
     7801 atattgaatt ttcaaaaatt cttacttttt ttttggatgg acgcaaagaa gtttaataat
     7861 catattacat ggcattacca ccatatacat atccatatac atatccatat ctaatcttac
     7921 ttatatgttg tggaaatgta aagagcccca ttatcttagc ctaaaaaaac cttctctttg
     7981 gaactttcag taatacgctt aactgctcat tgctatattg aagtgcggcc gcgtgggcgt
     8041 gcgtgggcgg gcgtgggcgt gcgtgggcgg gcgtgggcgt gcgtgggcgt ctagaccgtg
     8101 cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg tgcctcgcgc cgcactgctc
     8161 cgaacaataa agattctaca atactagctt ttatggttat gaagaggaaa aattggcagt
     8221 aacctggccc cacaaacctt caaattaacg aatcaaatta acaaccatag gatgataatg
     8281 cgattagttt tttagcctta tttctggggt aattaatcag cgaagcgatg atttttgatc
     8341 tattaacaga tatataaatg gaaaagctgc ataaccactt taactaatac tttcaacatt
     8401 ttcagtttgt attacttctt attcaaatgt cataaaagta tcaacaaaaa attgttaata
     8461 tacctctata ctttaacgtc aaggagaaaa aactataagg gtgcgcatct gatgagtccg
     8521 tgaggacgaa acgagctagc tcgtcatgcg cacccttagc gagaggttta tcattaaggt
     8581 caacctctgg atgttgtttc ggcatcctgc attgaatctg agttactgtc tgttttcctg
     8641 ggtcacgcgt aggagggttt tccattcgtc ttgaagtcga ggactttggc atacgatgga
     8701 agagcgtaac ttcgttgtaa agaataagga aatgattccg gaagctttgg aagtaaggaa
     8761 acccgtttct tctgacgtaa gggtgcgcaa ctttggcata cgatggaagg tttcagagct
     8821 atgctggaaa cagcatagca agttgaaata aggctagtcc gttatcaact tgaaaaagtg
     8881 gcaccgagtc ggtgcttttt gatggccggc atggtcccag cctcctcgct ggcgccggct
     8941 gggcaacacc ttcgggtggc gaatgggact taaagaaagt ggaatattca ttcatatcat
     9001 attttttcta ttaactgcct ggtttctttt aaatttttta ttggttgtcg acttgaacgg
     9061 agtgacaata tatatatata tatttaataa tgacatcatt atctgtaaat ctgattctta
     9121 atgctattct agttatgtaa gagtggtcct ttccat
//
