LOCUS       sequence-122738-        5688 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Improved version for complementation of yeast auxotrophic marker 
            TRP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5688)
  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 5688)
  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: LD-BS-Trp1
FEATURES             Location/Qualifiers
     source          1..5688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..21
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5..21
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     40..60
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        42..60
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        796..1073
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     1810..1826
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1811..1827
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1861..1877)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1861..1877)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2904..2923)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2905..2923)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2933..2950)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2933..2949)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2942..2964)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3091..3546
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3178..3197)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3388..3409
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3572..3676
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3677..3745,3746..4537)
                     /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(3895..3914)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     5208..5227
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5461..5478
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5595..5616
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5631..5636,5637..5654,5655..5661)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5669..5685
                     /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     join(5674..5688,1..8)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
       61 gaacaaaagc tggtactatt agctgaattg ccactgctat cgttgttagt ggcgttagtg
      121 cttgcattca aagacatgga gggcgttatt acgccggagc tcctcgacag cagatctgat
      181 gactggtcaa tatatttttg cattgaggct ctgtttggaa ttatattttg agatgaccca
      241 tctaatgtac tggtatcacc agatttcatg tcgtttttta aagcggctgc ttgagtctta
      301 gcaatagcgt caccatctgg tgaatccttt gaaggaacca ctgacgaagg tttggacagt
      361 gacgaagagg atctttcctg ctttgaatta gtcgcgctgg gagcagatga cgagttggtg
      421 gagctggggg caggattgct ggccgtcgtg ggtcctgaat gggtccttgg ctggtccatc
      481 tctattctga aaacggaaga ggagtaggga atattactgg ctgaaaataa gtcttgaatg
      541 aacgtatacg cgtatatttc taccaatctc tcaacactga gtaatggtag ttataagaaa
      601 gagaccgagt tagggacagt tagaggcggt ggagatattc cttatggcat gtctggcgat
      661 gataaaactt ttcaaacggc agccccgatc taaaagagct gacagggaaa tggtcagaaa
      721 aagaaacgtg cacccgcccg tctggacgcg ccgctcaccc gcacggcaga gaccaatcag
      781 taaaaatcaa cggttaacga cattactata tatataatat aggaagcatt taatagaaca
      841 gcatcgtaat atatgtgtac tttgcagtta tgacgccaga tggcagtagt ggaagatatt
      901 ctttattgaa aaatagcttg tcaccttacg tacaatcttg atccggagct tttctttttt
      961 tgccgattaa gaattcggtc gaaaaaagaa aaggagaggg ccaagaggga gggcattggt
     1021 gactattgag cacgtgagta tacgtgatta agcacacaaa ggcagcttgg agtatgctcg
     1081 ttaaagtgtg tggtttgcaa accgttgaag ctgcaaagac tgctgtggat gatggtgctg
     1141 attacttagg tatcatttgt gttcccggta ggaaaagaac cattgactca tctgttgcga
     1201 aaggtatttc aactgcagtt caccaacaag agaacgtgaa aggtactaaa ctagtcgggg
     1261 tgtttagaaa tcagtccgtt gatgatgtcc ttcaactgta ccacgaatat aatctagatg
     1321 tgatacaatt acatggagat gaagatatta aagaatacag atctttgatc ccatcttcaa
     1381 ttccaatcat taagaggttc cagttcccac aggattgtga attactactg gacctgtatg
     1441 aacacgtaga caatgtgctg acgttgttcg attctggtga aggtggcact ggtgagaaat
     1501 tgaattggag tgcaatttcc agttggtctg caagtcatcc cgagataaaa ttcattatcg
     1561 ctggtggatt gaatcctgat aacgtttctg ttgccattaa tatgttacca aatgcgatcg
     1621 gtgtcgatgt aagtggagga gtagagactg atggtatcaa ggatttagaa aaggtaaagc
     1681 tattcatcca gcaggcctct caatagagga aaaaacgtat ctttcttcaa ttacacatac
     1741 gtatatataa attgatatat atatagtgaa tgatcgttcc gcggccgctc tagggtaccg
     1801 ggccccccct cgaggtcgac ggtatcgata agcttgatat cgaattcctg cagcccgggg
     1861 gatccactag ttctagagcg gccgccaccg cggtgggtta ttactgagta gtatttattt
     1921 aagtattgtt tgtgcacttg cctgcaggcc ttttgaaaag caagcataaa agatctaaac
     1981 ataaaatctg taaaataaca agatgtaaag ataatgctaa atcatttggc tttttgattg
     2041 attgtacagg aaaatataca tcgcaggggg ttgactttta ccatttcacc gcaatggaat
     2101 caaacttgtt gaagagaatg ttcacaggcg catacgctac aatgacccga ttcttgctag
     2161 ccttttctcg gtcttgcaaa caaccgccgg cagcttagta tataaataca catgtacata
     2221 cctctctccg tatcctcgta atcattttct tgtatttatc gtcttttcgc tgtaaaaact
     2281 ttatcacact tatctcaaat acacttatta accgctttta ctattatctt ctacgctgac
     2341 agtaatatca aacagtgaca catattaaac acagtggttt ctttgcataa acaccatcag
     2401 cctcaagtcg tcaagtaaag atttcgtgtt catgcagata gataacaatc tatatgttga
     2461 taattagcgt tgcctcatca atgcgagatc cgtttaaccg gaccctagtg cacttacccc
     2521 acgttcggtc cactgtgtgc cgaacatgct ccttcactat tttaacatgt ggaattcttg
     2581 aaagaatgaa atcgccatgc caagccatca cacggtcttt tatgcaattg attgaccgcc
     2641 tgcaacacat aggcagtaaa atttttactg aaacgtatat aatcatcata agcgacaagt
     2701 gaggcaacac ctttgttacc acattgacaa ccccaggtat tcatacttcc tattagcgga
     2761 atcaggagtg caaaaagaga aaataaaagt aaaaaggtag ggcaacacat agtatgaata
     2821 caaacgttcc aatattcagt tctccggtca gagatttacc aaggtctttc gaacaaaaac
     2881 atttagcggt tgtagacaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
     2941 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     3001 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     3061 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     3121 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3181 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3241 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3301 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3361 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3421 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3481 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     3541 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3601 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3661 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3721 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3781 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3841 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3901 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3961 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4021 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4081 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4141 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4201 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     4261 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4321 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4381 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4441 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4501 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4561 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4621 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4681 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4741 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4801 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     4861 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4921 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4981 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5041 cacacagccc agcttggagc gaacgaccta caccgaacaa aagtgccact gagataccta
     5101 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5161 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5221 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5281 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5341 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5401 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     5461 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     5521 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     5581 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     5641 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaattt
//
