LOCUS       sequence-121985-        4716 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  complementation of yeast auxotrophic marker HIS3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4716)
  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 4716)
  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-His3
FEATURES             Location/Qualifiers
     source          1..4716
                     /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"
     promoter        798..985
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             986..1648
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     primer_bind     complement(2475..2491)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2475..2491)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2528..2548)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2528..2546)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2567..2583)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2567..2583)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2580..2602)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2591..2607
                     /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(2615..2621,2622..2639,2640..2645))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2660..2681
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2798..2815)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2969..3557)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3049..3068)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3728..4519,4520..4588))
                     /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     4351..4370
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4589..4693)
                     /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 gaggtcgaca agagcttggt gagcgctagg agtcactgcc aggtatcgtt
      721 tgaacacggc attagtcagg gaagtcataa cacagtcctt tcccgcaatt ttctttttct
      781 attactcttg gcctcctcta gtacactcta tattttttta tgcctcggta atgattttca
      841 tttttttttt tccacctagc ggatgactct ttttttttct tagcgattgg cattatcaca
      901 taatgaatta tacattatat aaagtaatgt gatttcttcg aagaatatac taaaaaatga
      961 gcaggcaaga taaacgaagg caaagatgac agagcagaaa gccctagtaa agcgtattac
     1021 aaatgaaacc aagattcaga ttgcgatctc tttaaagggt ggtcccctag cgatagagca
     1081 ctcgatcttc ccagaaaaag aggcagaagc agtagcagaa caggccacac aatcgcaagt
     1141 gattaacgtc cacacaggta tagggtttct ggaccatatg atacatgctc tggccaagca
     1201 ttccggctgg tcgctaatcg ttgagtgcat tggtgactta cacatagacg accatcacac
     1261 cactgaagac tgcgggattg ctctcggtca agcttttaaa gaggccctag gggccgtgcg
     1321 tggagtaaaa aggtttggat caggatttgc gcctttggat gaggcacttt ccagagcggt
     1381 ggtagatctt tcgaacaggc cgtacgcagt tgtcgaactt ggtttgcaaa gggagaaagt
     1441 aggagatctc tcttgcgaga tgatcccgca ttttcttgaa agctttgcag aggctagcag
     1501 aattaccctc cacgttgatt gtctgcgagg caagaatgat catcaccgta gtgagagtgc
     1561 gttcaaggct cttgcggttg ccataagaga agccacctcg cccaatggta ccaacgatgt
     1621 tccctccacc aaaggtgttc ttatgtagtg acaccgatta tttaaagctg cagcatacga
     1681 tatatataca tgtgtatata tgtataccta tgaatgtcag taagtatgta tacgaacagt
     1741 atgatactga agatgacaag gtaatgcatc attctatacg tgtcattctg aacgaggcgc
     1801 gctttccttt tttctttttg ctttttcttt ttttttctct tgaactcgag aaaaaaaata
     1861 taaaagagat ggaggaacgg gaaaaagtta gttgtggtga taggtggcaa gtggtattcc
     1921 gtaagaacaa caagaaaagc atttcatatt atggctgaac tgagcgaaca agtgcaaaat
     1981 ttaagcatca acgacaacaa cgagaatggt tatgttcctc ctcacttaag aggaaaacca
     2041 agaagtgcca gaaataacag tagcaactac aataacaaca acggcggcta caacggtggc
     2101 cgtggcggtg gcagcttctt tagcaacaac cgtcgtggtg gttacggcaa cggtggtttc
     2161 ttcggtggaa acaacggtgg cagcagatct aacggccgtt ctggtggtag atggatcgat
     2221 ggcaaacatg tcccagctcc aagaaacgaa aaggccgaga tcgccatatt tggtgtcccc
     2281 gaggatccaa atttccaatc ttctggtatt aacttcgata actacgatga tattccagtg
     2341 gacgcctctg gtaaggatgt tcctgaacca atcacagaat ttacctcacc tccattggac
     2401 ggattgttat tggaaaacat caaattggcc cgtttcacca agccaacacc tgtgcgaatt
     2461 cctgcagccc gggggatcca ctagttctag agcggccgcc accgcggtgg agctccagct
     2521 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
     2581 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     2641 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     2701 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2761 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2821 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2881 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2941 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3001 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3061 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3121 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3181 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3241 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3301 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3361 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     3421 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3481 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3541 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3601 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3661 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3721 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3781 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3841 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3901 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3961 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4021 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4081 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4141 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4201 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4261 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4321 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4381 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4441 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4501 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4561 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4621 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4681 taggggttcc gcgcacattt ccccgaaaag tgccac
//
