LOCUS       sequence-121984-        5330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  complementation of yeast auxotrophic marker TRP1. Although this is 
            the plasmid described in the paper, the depositor notes that it has 
            a low integration efficiency and Plasmid 69503 should be used. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5330)
  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 5330)
  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-Trp1
FEATURES             Location/Qualifiers
     source          1..5330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(129..197,198..989)
                     /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     complement(347..366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1649..1668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1902..1919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2072..2077,2078..2095,2096..2102)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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)."
     primer_bind     2115..2137
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2134..2150
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2169..2189
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2260..2537
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             2538..3212
                     /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"
     rep_origin      3051..3888
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     primer_bind     complement(4686..4705)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4687..4705)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4715..4732)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4715..4731)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4724..4746)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4873..5328
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4960..4979)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5170..5191
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctccac cgcggtggcg
     2221 gccgctctag aactagtcaa tcagtaaaaa tcaacggtta acgacattac tatatatata
     2281 atataggaag catttaatag aacagcatcg taatatatgt gtactttgca gttatgacgc
     2341 cagatggcag tagtggaaga tattctttat tgaaaaatag cttgtcacct tacgtacaat
     2401 cttgatccgg agcttttctt tttttgccga ttaagaattc ggtcgaaaaa agaaaaggag
     2461 agggccaaga gggagggcat tggtgactat tgagcacgtg agtatacgtg attaagcaca
     2521 caaaggcagc ttggagtatg tctgttatta atttcacagg tagttctggt ccattggtga
     2581 aagtttgcgg cttgcagagc acagaggccg cagaatgtgc tctagattcc gatgctgact
     2641 tgctgggtat tatatgtgtg cccaatagaa agagaacaat tgacccggtt attgcaagga
     2701 aaatttcaag tcttgtaaaa gcatataaaa atagttcagg cactccgaaa tacttggttg
     2761 gcgtgtttcg taatcaacct aaggaggatg ttttggctct ggtcaatgat tacggcattg
     2821 atatcgtcca actgcatgga gatgagtcgt ggcaagaata ccaagagttc ctcggtttgc
     2881 cagttattaa aagactcgta tttccaaaag actgcaacat actactcagt gcagcttcac
     2941 agaaacctca ttcgtttatt cccttgtttg attcagaagc aggtgggaca ggtgaacttt
     3001 tggattggaa ctcgatttct gactgggttg gaaggcaaga gagccccgaa agcttacatt
     3061 ttatgttagc tggtggactg acgccagaaa atgttggtga tgcgcttaga ttaaatggcg
     3121 ttattggtgt tgatgtaagc ggaggtgtgg agacaaatgg tgtaaaagac tctaacaaaa
     3181 tagcaaattt cgtcaaaaat gctaagaaat aggttattac tgagtagtat ttatttaagt
     3241 attgtttgtg cacttgcctg caggcctttt gaaaagcaag cataaaagat ctaaacataa
     3301 aatctgtaaa ataacaagat gtaaagataa tgctaaatca tttggctttt tgattgattg
     3361 tacaggaaaa tatacatcgc agggggttga cttttaccat ttcaccgcaa tggaatcaaa
     3421 cttgttgaag agaatgttca caggcgcata cgctacaatg acccgattct tgctagcctt
     3481 ttctcggtct tgcaaacaac cgccggcagc ttagtatata aatacacatg tacatacctc
     3541 tctccgtatc ctcgtaatca ttttcttgta tttatcgtct tttcgctgta aaaactttat
     3601 cacacttatc tcaaatacac ttattaaccg cttttactat tatcttctac gctgacagta
     3661 atatcaaaca gtgacacata ttaaacacag tggtttcttt gcataaacac catcagcctc
     3721 aagtcgtcaa gtaaagattt cgtgttcatg cagatagata acaatctata tgttgataat
     3781 tagcgttgcc tcatcaatgc gagatccgtt taaccggacc ctagtgcact taccccacgt
     3841 tcggtccact gtgtgccgaa catgctcctt cactatttta acatgtggaa ttcttgaaag
     3901 aatgaaatcg ccatgccaag ccatcacacg gtcttttatg caattgattg accgcctgca
     3961 acacataggc agtaaaattt ttactgaaac gtatataatc atcataagcg acaagtgagg
     4021 caacaccttt gttaccacat tgacaacccc aggtattcat acttcctatt agcggaatca
     4081 ggagtgcaaa aagagaaaat aaaagtaaaa aggtagggca acacatagta tgaatacaaa
     4141 cgttccaata ttcagttctc cggtcagaga tttaccaagg tctttcgaac aaaaacattt
     4201 agcggttgta gatgcatttt tccaaacata ccatgtcaaa cctgatttta tcgctaggtc
     4261 tcctggcaga gtaaatctga ttggtgagca tatagattat tgcgattttt cagttttgcc
     4321 attagccatt gatgtggata tgctttgcgc agttaaaatt ttagacgaaa aaaatccatc
     4381 cattacctta acaaatgcgg accctaaatt tgctcagcga aagtttgatc tgcctttaga
     4441 tggttcctac atggccatag atccgtctgt gtcggaatgg tcgaattact ttaaatgcgg
     4501 actacatgtg gcacattcat acttgaaaaa aattgctccg gaaagattta ataatacacc
     4561 cttagtaggt gcgcagatct tttgccagag cgatattcct actggtggtg gactctcatc
     4621 tgcatttact tgcgcggcag cactagccac aattagactc gagggggggc ccggtaccca
     4681 attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg
     4741 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     4801 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     4861 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4921 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4981 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5041 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     5101 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5161 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     5221 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     5281 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
//
