LOCUS       sequence-128693-        8338 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  complementation of yeast auxotrophic marker LYS2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8338)
  AUTHORS   Li D, Song JZ, Shan MH, Li SP, Liu W, Li H, Zhu J, Wang Y, Lin J, 
            Xie Z
  TITLE     A fluorescent tool set for yeast Atg proteins.
  JOURNAL   Autophagy. 2015;11(6):954-60. doi: 10.1080/15548627.2015.1040971.
  PUBMED    25998947
REFERENCE   2  (bases 1 to 8338)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Autophagy."; date: "2015"; volume: "11(6):954-60. doi"; pages: " 
            10.1080/15548627.2015.1040971"
COMMENT     Sequence Label: BS-Lys2
FEATURES             Location/Qualifiers
     source          1..8338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1456..1472)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1456..1472)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        1523..1762
                     /gene="S. cerevisiae LYS2"
                     /label=LYS2 promoter
     CDS             1763..5941
                     /codon_start=1
                     /gene="S. cerevisiae LYS2"
                     /product="alpha-aminoadipate reductase, required for lysine
                     biosynthesis"
                     /label=LYS2
                     /note="yeast auxotrophic marker"
                     /translation="MTNEKVWIEKLDNPTLSVLPHDFLRPQQEPYTKQATYSLQLPQLD
                     VPHDSFSNKYAVALSVWAALIYRVTGDDDIVLYIANNKILRFNIQPTWSFNELYSTINN
                     ELNKLNSIEANFSFDELAEKIQSCQDLERTPQLFRLAFLENQDFKLDEFKHHLVDFALN
                     LDTSNNAHVLNLIYNSLLYSNERVTIVADQFTQYLTAALSDPSNCITKISLITASSKDS
                     LPDPTKNLGWCDFVGCIHDIFQDNAEAFPERTCVVETPTLNSDKSRSFTYRDINRTSNI
                     VAHYLIKTGIKRGDVVMIYSSRGVDLMVCVMGVLKAGATFSVIDPAYPPARQTIYLGVA
                     KPRGLIVIRAAGQLDQLVEDYINDELEIVSRINSIAIQENGTIEGGKLDNGEDVLAPYD
                     HYKDTRTGVVVGPDSNPTLSFTSGSEGIPKGVLGRHFSLAYYFNWMSKRFNLTENDKFT
                     MLSGIAHDPIQRDMFTPLFLGAQLYVPTQDDIGTPGRLAEWMSKYGCTVTHLTPAMGQL
                     LTAQATTPFPKLHHAFFVGDILTKRDCLRLQTLAENCRIVNMYGTTETQRAVSYFEVKS
                     KNDDPNFLKKLKDVMPAGKGMLNVQLLVVNRNDRTQICGIGEIGEIYVRAGGLAEGYRG
                     LPELNKEKFVNNWFVEKDHWNYLDKDNGEPWRQFWLGPRDRLYRTGDLGRYLPNGDCEC
                     CGRADDQVKIRGFRIELGEIDTHISQHPLVRENITLVRKNADNEPTLITFMVPRFDKPD
                     DLSKFQSDVPKEVETDPIVKGLIGYHLLSKDIRTFLKKRLASYAMPSLIVVMDKLPLNP
                     NGKVDKPKLQFPTPKQLNLVAENTVSETDDSQFTNVEREVRDLWLSILPTKPASVSPDD
                     SFFDLGGHSILATKMIFTLKKKLQVDLPLGTIFKYPTIKAFAAEIDRIKSSGGSSQGEV
                     VENVTANYAEDAKKLVETLPSSYPSREYFVEPNSAEGKTTINVFVTGVTGFLGSYILAD
                     LLGRSPKNYSFKVFAHVRAKDEEAAFARLQKAGITYGTWNEKFASNIKVVLGDLSKSQF
                     GLSDEKWMDLANTVDIIIHNGALVHWVYPYAKLRDPNVISTINVMSLAAVGKPKFFDFV
                     SSTSTLDTEYYFNLSDKLVSEGKPGILESDDLMNSASGLTGGYGQSKWAAEYIIRRAGE
                     RGLRGCIVRPGYVTGASANGSSNTDDFLLRFLKGSVQLGKIPDIENSVNMVPVDHVARV
                     VVATSLNPPKENELAVAQVTGHPRILFKDYLYTLHDYGYDVEIESYSKWKKSLEASVID
                     RNEENALYPLLHMVLDNLPESTKAPELDDRNAVASLKKDTAWTGVDWSNGIGVTPEEVG
                     IYIAFLNKVGFLPPPTHNDKLPLPSIELTQAQISLVASGAGARGSSAAA"
     primer_bind     complement(6150..6170)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6150..6168)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6189..6205)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6189..6205)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6202..6224)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6213..6229
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6237..6243,6244..6261,6262..6267))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6282..6303
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6420..6437)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6591..7179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6671..6690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7350..8141,8142..8210))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7973..7992
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8211..8315)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggaatatg ttcatagggt agacgaaact atatacgcaa tctacataca
      721 tttatcaaga aggagaaaaa ggaggatgta aaggaataca ggtaagcaaa ttgatactaa
      781 tggctcaacg tgataaggaa aaagaattgc actttaacat taatattgac aaggaggagg
      841 gcaccacaca aaaagttagg tgtaacagaa aatcatgaaa ctatgattcc taatttatat
      901 attggaggat tttctctaaa aaaaaaaaaa tacaacaaat aaaaaacact caatgacctg
      961 accatttgat ggagtttaag tcaatacctt cttgaaccat ttcccataat ggtgaaagtt
     1021 ccctcaagaa ttttactctg tcagaaacgg ccttaacgac gtagtcgacc tcctcttcag
     1081 tactaaatct accaatacca aatctgatgg aagaatgggc taatgcatca tccttaccca
     1141 gcgcatgtaa aacataagaa ggttctaggg aagcagatgt acaggctgaa cccgaggata
     1201 atgcgatatc ccttagtgcc atcaataaag attctccttc cacgtaggcg aaagaaacgt
     1261 taacacaccc tggataacga tgatctggag atccgttcaa cgtggtatgt tcagcggata
     1321 atagaccttt gactaattta tcggatagtc ttttgatgtg agcttggtcg ttgtcaaatt
     1381 ctttcttcat caatctcgca gcttcaccaa atcccgctac caataagctt gatatcgaat
     1441 tcctgcagcc cgggggatcc actagttcta gaggcatcgc acagttttag cgaggaaaac
     1501 tcttcaatag ttttgccagc ggaattccac ttgcaattac ataaaaaatt ccggcggttt
     1561 ttcgcgtgtg actcaatgtc gaaatacctg cctaatgaac atgaacatcg cccaaatgta
     1621 tttgaagacc cgctgggaga agttcaagat atataagtaa caagcagcca atagtataaa
     1681 aaaaaatctg agtttattac ctttcctgga atttcagtga aaaactgcta attatagaga
     1741 gatatcacag agttactcac taatgactaa cgaaaaggtc tggatagaga agttggataa
     1801 tccaactctt tcagtgttac cacatgactt tttacgccca caacaagaac cttatacgaa
     1861 acaagctaca tattcgttac agctacctca gctcgatgtg cctcatgata gtttttctaa
     1921 caaatacgct gtcgctttga gtgtatgggc tgcattgata tatagagtaa ccggtgacga
     1981 tgatattgtt ctttatattg cgaataacaa aatcttaaga ttcaatattc aaccaacgtg
     2041 gtcatttaat gagctgtatt ctacaattaa caatgagttg aacaagctca attctattga
     2101 ggccaatttt tcctttgacg agctagctga aaaaattcaa agttgccaag atctggaaag
     2161 gacccctcag ttgttccgtt tggccttttt ggaaaaccaa gatttcaaat tagacgagtt
     2221 caagcatcat ttagtggact ttgctttgaa tttggatacc agtaataatg cgcatgtttt
     2281 gaacttaatt tataacagct tactgtattc gaatgaaaga gtaaccattg ttgcggacca
     2341 atttactcaa tatttgactg ctgcgctaag cgatccatcc aattgcataa ctaaaatctc
     2401 tctgatcacc gcatcatcca aggatagttt acctgatcca actaagaact tgggctggtg
     2461 cgatttcgtg gggtgtattc acgacatttt ccaggacaat gctgaagcct tcccagagag
     2521 aacctgtgtt gtggagactc caacactaaa ttccgacaag tcccgttctt tcacttatcg
     2581 cgacatcaac cgcacttcta acatagttgc ccattatttg attaaaacag gtatcaaaag
     2641 aggtgatgta gtgatgatct attcttctag gggtgtggat ttgatggtat gtgtgatggg
     2701 tgtcttgaaa gccggcgcaa ccttttcagt tatcgaccct gcatatcccc cagccagaca
     2761 aaccatttac ttaggtgttg ctaaaccacg tgggttgatt gttattagag ctgctggaca
     2821 attggatcaa ctagtagaag attacatcaa tgatgaattg gagattgttt caagaatcaa
     2881 ttccatcgct attcaagaaa atggtaccat tgaaggtggc aaattggaca atggcgagga
     2941 tgttttggct ccatatgatc actacaaaga caccagaaca ggtgttgtag ttggaccaga
     3001 ttccaaccca accctatctt tcacatctgg ttccgaaggt attcctaagg gtgttcttgg
     3061 tagacatttt tccttggctt attatttcaa ttggatgtcc aaaaggttca acttaacaga
     3121 aaatgataaa ttcacaatgc tgagcggtat tgcacatgat ccaattcaaa gagatatgtt
     3181 tacaccatta tttttaggtg cccaattgta tgtccctact caagatgata ttggtacacc
     3241 gggccgttta gcggaatgga tgagtaagta tggttgcaca gttacccatt taacacctgc
     3301 catgggtcaa ttacttactg cccaagctac tacaccattc cctaagttac atcatgcgtt
     3361 ctttgtgggt gacattttaa caaaacgtga ttgtctgagg ttacaaacct tggcagaaaa
     3421 ttgccgtatt gttaatatgt acggtaccac tgaaacacag cgtgcagttt cttatttcga
     3481 agttaaatca aaaaatgacg atccaaactt tttgaaaaaa ttgaaagatg tcatgcctgc
     3541 tggtaaaggt atgttgaacg ttcagctact agttgttaac aggaacgatc gtactcaaat
     3601 atgtggtatt ggcgaaatag gtgagattta tgttcgtgca ggtggtttgg ccgaaggtta
     3661 tagaggatta ccagaattga ataaagaaaa atttgtgaac aactggtttg ttgaaaaaga
     3721 tcactggaat tatttggata aggataatgg tgaaccttgg agacaattct ggttaggtcc
     3781 aagagataga ttgtacagaa cgggtgattt aggtcgttat ctaccaaacg gtgactgtga
     3841 atgttgcggt agggctgatg atcaagttaa aattcgtggg ttcagaatcg aattaggaga
     3901 aatagatacg cacatttccc aacatccatt ggtaagagaa aacattactt tagttcgcaa
     3961 aaatgccgac aatgagccaa cattgatcac atttatggtc ccaagatttg acaagccaga
     4021 tgacttgtct aagttccaaa gtgatgttcc aaaggaggtt gaaactgacc ctatagttaa
     4081 gggcttaatc ggttaccatc ttttatccaa ggacatcagg actttcttaa agaaaagatt
     4141 ggctagctat gctatgcctt ccttgattgt ggttatggat aaactaccat tgaatccaaa
     4201 tggtaaagtt gataagccta aacttcaatt cccaactccc aagcaattaa atttggtagc
     4261 tgaaaataca gtttctgaaa ctgacgactc tcagtttacc aatgttgagc gcgaggttag
     4321 agacttatgg ttaagtatat tacctaccaa gccagcatct gtatcaccag atgattcgtt
     4381 tttcgattta ggtggtcatt ctatcttggc taccaaaatg atttttacct taaagaaaaa
     4441 gctgcaagtt gatttaccat tgggcacaat tttcaagtat ccaacgataa aggcctttgc
     4501 cgcggaaatt gacagaatta aatcatcggg tggatcatct caaggtgagg tcgtcgaaaa
     4561 tgtcactgca aattatgcgg aagacgccaa gaaattggtt gagacgctac caagttcgta
     4621 cccctctcga gaatattttg ttgaacctaa tagtgccgaa ggaaaaacaa caattaatgt
     4681 gtttgttacc ggtgtcacag gatttctggg ctcctacatc cttgcagatt tgttaggacg
     4741 ttctccaaag aactacagtt tcaaagtgtt tgcccacgtc agggccaagg atgaagaagc
     4801 tgcatttgca agattacaaa aggcaggtat cacctatggt acttggaacg aaaaatttgc
     4861 ctcaaatatt aaagttgtat taggcgattt atctaaaagc caatttggtc tttcagatga
     4921 gaagtggatg gatttggcaa acacagttga tataattatc cataatggtg cgttagttca
     4981 ctgggtttat ccatatgcca aattgaggga tccaaatgtt atttcaacta tcaatgttat
     5041 gagcttagcc gccgtcggca agccaaagtt ctttgacttt gtttcctcca cttctactct
     5101 tgacactgaa tactacttta atttgtcaga taaacttgtt agcgaaggga agccaggcat
     5161 tttagaatca gacgatttaa tgaactctgc aagcgggctc actggtggat atggtcagtc
     5221 caaatgggct gctgagtaca tcattagacg tgcaggtgaa aggggcctac gtgggtgtat
     5281 tgtcagacca ggttacgtaa caggtgcctc tgccaatggt tcttcaaaca cagatgattt
     5341 cttattgaga tttttgaaag gttcagtcca attaggtaag attccagata tcgaaaattc
     5401 cgtgaatatg gttccagtag atcatgttgc tcgtgttgtt gttgctacgt ctttgaatcc
     5461 tcccaaagaa aatgaattgg ccgttgctca agtaacgggt cacccaagaa tattattcaa
     5521 agactacttg tatactttac acgattatgg ttacgatgtc gaaatcgaaa gctattctaa
     5581 atggaagaaa tcattggagg cgtctgttat tgacaggaat gaagaaaatg cgttgtatcc
     5641 tttgctacac atggtcttag acaacttacc tgaaagtacc aaagctccgg aactagacga
     5701 taggaacgcc gtggcatctt taaagaaaga caccgcatgg acaggtgttg attggtctaa
     5761 tggaataggt gttactccag aagaggttgg tatatatatt gcatttttaa acaaggttgg
     5821 atttttacct ccaccaactc ataatgacaa acttccactg ccaagtatag aactaactca
     5881 agcgcaaata agtctagttg cttcaggtgc tggtgctcgt ggaagctccg cagcagctta
     5941 aggttgagca ttacgtatga tatgtccatg tacaataatt aaatatgaat taggagaaag
     6001 acttagcttc ttttcgggtg atgtcactta aaaactccga gaataatata taataagaga
     6061 ataaaatatt agttattgaa taagaactgt aaatcagctg gcgttagtct gctgcggccg
     6121 ccaccgcggt ggagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     6181 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6241 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6301 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6361 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6421 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6481 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6541 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6601 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6661 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6721 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6781 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6841 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6901 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6961 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7021 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7081 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7141 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7201 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7261 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7321 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7381 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7441 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7501 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7561 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7621 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7681 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7741 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7801 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7861 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7921 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7981 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     8041 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     8101 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     8161 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8221 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8281 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccac
//
