LOCUS       sequence-121989-        6782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RFP-Atg8 (PAS  marker) with  improved  property.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6782)
  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 6782)
  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: PCUP1-DuDre-Atg8-404
FEATURES             Location/Qualifiers
     source          1..6782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(542..997)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(679..700)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     891..910
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1127..1149
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1141..1158
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1142..1158
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1168..1186
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1451..1685
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     primer_bind     complement(3823..3843)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3823..3841)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3862..3878)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3862..3878)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3875..3897)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3886..3902
                     /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(3910..3916,3917..3934,3935..3940))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4093..4110)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4264..4852)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4344..4363)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5023..5814,5815..5883))
                     /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     5646..5665
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5884..5988)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6056..6074
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6112..6134)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6234..6253
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6270..6550
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             join(6551..6782,1..443)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
ORIGIN
        1 gtaatcaacc taaggaggat gttttggctc tggtcaatga ttacggcatt gatatcgtcc
       61 aactgcatgg agatgagtcg tggcaagaat accaagagtt cctcggtttg ccagttatta
      121 aaagactcgt atttccaaaa gactgcaaca tactactcag tgcagcttca cagaaacctc
      181 attcgtttat tcccttgttt gattcagaag caggtgggac aggtgaactt ttggattgga
      241 actcgatttc tgactgggtt ggaaggcaag agagccccga aagcttacat tttatgttag
      301 ctggtggact gacgccagaa aatgttggtg atgcgcttag attaaatggc gttattggtg
      361 ttgatgtaag cggaggtgtg gagacaaatg gtgtaaaaga ctctaacaaa atagcaaatt
      421 tcgtcaaaaa tgctaagaaa taggttatta ctgagtagta tttatttaag tattgtttgt
      481 gcacttgcct atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
      541 gaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
      601 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
      661 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
      721 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
      781 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
      841 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
      901 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
      961 acacccgccg cgcttaatgc gccgctacag ggcgcgtcgc gccattcgcc attcaggctg
     1021 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     1081 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     1141 tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc
     1201 gggccccccc tcgaggtcga catctgggta ttactacggc aaacttcaac gatttctatg
     1261 atgcatttta taattagtaa gccgatccca ttaccgacat ttgggcgcta tacgtgcata
     1321 tgttcatgta tgtatctgta tttaaaacac ttttgtatta tttttcctca tatatgtgta
     1381 taggtttata cggatgattt aattattact tcaccaccct ttatttcagg ctgatatctt
     1441 agccttgtta ctagttagaa aaagacattt ttgctgtcag tcactgtcaa gagattcttt
     1501 tgctggcatt tcttctagaa gcaaaaagag cgatgcgtct tttccgctga accgttccag
     1561 caaaaaagac taccaacgca atatggattg tcagaatcat ataaaagaga agcaaataac
     1621 tccttgtctt gtatcaattg cattataata tcttcttgtt agtgcaatat catatagaag
     1681 tcatcgaaat agatattaag aaaaacaaac tgtacaatca atcaatcaat catcacataa
     1741 aatgtctgaa ttcatggttt ccaaaggtga agagaacaat atggctgtca tcaagccctt
     1801 catgagattc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     1861 cgagggcgag ggcaagccct acgagggcac ccagaccgcc aagctgcagg tgaccaaggg
     1921 cggccccctg cccttcgcct gggacatcct gtccccccag ttccagtacg gctccaaggt
     1981 gtacgtgaag caccccgccg acatccccga ctacaagaag ctgtccttcc ccgagggctt
     2041 caagtgggag agagtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     2101 ctccctgcag gacggcacct tcatctacca cgtgaagttc atcggcgtga acttcccctc
     2161 cgacggcccc gtaatgcaga agaagactct gggctgggag ccctccaccg agagactgta
     2221 ccccagagac ggcgtgctga agggcgagat ccacaaggcg ctgaagctga agggcggcgg
     2281 ccactacctg gtggagttca agtcaatcta catggccaag aagcccgtga agctgcccgg
     2341 ctactactac gtggactcca agctggacat cacctcccac aacgaggact acaccgtggt
     2401 ggagcagtac gagagagccg aggccagaca ccacctgttc ttaattccag ttccaacgcg
     2461 tgatcttgag aagccattca acgtcatcaa gcccttcatg cgcttcaagg tgcacatgga
     2521 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggca agccctacga
     2581 gggcacccag accgccaagc tgcaggtgac caagggcggc cccctgccct tcgcctggga
     2641 catcctgtcc ccccagttcc agtacggctc caaggtgtac gtgaagcacc ccgccgacat
     2701 ccccgactac aagaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     2761 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcaccttcat
     2821 ctaccacgtg aagttcatcg gcgtgaactt cccctccgac ggccccgtaa tgcagaagaa
     2881 gactctgggc tgggagccct ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg
     2941 cgagatccac aaggcgctga agctgaaggg cggcggccac tacctggtgg agttcaagtc
     3001 aatctacatg gccaagaagc ccgtgaagct gcccggctac tactacgtgg actccaagct
     3061 ggacatcacc tcccacaacg aggactacac cgtggtggag cagtacgagc gcgccgaggc
     3121 ccgccaccac ctgttcttaa ttgtcgttct gaacagagct gagactccat tgccattgga
     3181 tccaaccggt aaggtcaagg ctgagctgga tactaggatg aagtctacat ttaagtctga
     3241 atatccattt gaaaaaagga aggcggagtc ggagaggatt gctgacaggt tcaagaatag
     3301 gatacctgtg atttgcgaaa aagctgaaaa gtcagatatt ccagagattg ataagcgtaa
     3361 atatctagtt cctgctgacc ttaccgtagg gcaatttgtt tatgttataa gaaagagaat
     3421 tatgctaccc cctgagaagg ccatcttcat ttttgtcaat gatactttgc cacctactgc
     3481 ggcgttgatg tctgccatat atcaagaaca caaggataag gacgggtttt tgtatgtcac
     3541 ttactcagga gaaaatacat ttggcaggta gtcttttata tgaaaagaaa tgaagcgtta
     3601 acatctaaaa tcgaaattat agggactcat tagccatcta ttgttccagt ataagaatgt
     3661 ttacataaaa aattcgaaaa aaaaagataa aataagggaa aaatatatat atattcatta
     3721 agaagttttg tacacaacgg agccaagatt tcaccagtat ctcgaggggg ggcccggtac
     3781 ccaattcgcc ctatagtgag tcgtgagctc cagcttttgt tccctttagt gagggttaat
     3841 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     3901 aattccacac aacataggag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3961 gaggtaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4021 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4081 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4141 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4201 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4261 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4321 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4381 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4441 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4501 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4561 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4621 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4681 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     4741 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4801 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4861 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4921 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4981 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5041 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5101 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5161 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5221 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5281 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5341 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5401 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5461 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5521 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5581 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5641 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5701 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5761 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5821 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5881 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5941 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6001 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     6061 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     6121 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     6181 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     6241 agagcagatt gtactgagag tgcaccataa acgacattac tatatatata atataggaag
     6301 catttaatag acagcatcgt aatatatgtg tactttgcag ttatgacgcc agatggcagt
     6361 agtggaagat attctttatt gaaaaatagc ttgtcacctt acgtacaatc ttgatccgga
     6421 gcttttcttt ttttgccgat taagaattaa ttcggtcgaa aaaagaaaag gagagggcca
     6481 agagggaggg cattggtgac tattgagcac gtgagtatac gtgattaagc acacaaaggc
     6541 agcttggagt atgtctgtta ttaatttcac aggtagttct ggtccattgg tgaaagtttg
     6601 cggcttgcag agcacagagg ccgcagaatg tgctctagat tccgatgctg acttgctggg
     6661 tattatatgt gtgcccaata gaaagagaac aattgacccg gttattgcaa ggaaaatttc
     6721 aagtcttgta aaagcatata aaaatagttc aggcactccg aaatacttgg ttggcgtgtt
     6781 tc
//
