LOCUS       sequence-430366-        7546 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DsRed-Express 2 as a marker of cytosol, uses auxotrophic 
            marker URA3(Saccharomyces cerevisiae)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7546)
  AUTHORS   He CW, Cui XF, Ma SJ, Xu Q, Ran YP, Chen WZ, Mu JX, Li H, Zhu J, 
            Gong Q, Xie Z
  TITLE     Membrane recruitment of Atg8 by Hfl1 facilitates turnover of 
            vacuolar membrane proteins in yeast cells approaching stationary 
            phase.
  JOURNAL   BMC Biol. 2021 Jun 4;19(1):117. doi: 10.1186/s12915-021-01048-7.
  PUBMED    34088313
REFERENCE   2  (bases 1 to 7546)
  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: "BMC Biol.";
            date: "2021-06-4"; volume: "19(1):117. doi"; pages: " 
            10.1186/s12915-021-01048-7"
COMMENT     Sequence Label: PTPI1-Dsred-URA3
FEATURES             Location/Qualifiers
     source          1..7546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(39..59)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     257..276
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     451..474
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(574..590)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(574..590)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1582..1602)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1582..1600)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1621..1637)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1621..1637)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1634..1656)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1645..1661
                     /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(1669..1675,1676..1693,1694..1699))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1714..1735
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1852..1869)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2023..2611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2103..2122)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2782..3573,3574..3642))
                     /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     3405..3424
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3643..3747)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3784..4287
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4328..4346
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4384..4406)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4506..4525
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4549..4764
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4765..5568
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(5699..6154)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5836..5857)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6048..6067
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6280..6302
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6294..6311
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6295..6311
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6321..6340
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6321..6339
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        6846..7365
                     /gene="S. cerevisiae TPI1"
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose 
                     phosphate isomerase"
ORIGIN
        1 tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag ttccagtacg
       61 gctccaaggt gtacgtgaag caccccgccg acatccccga ctacaagaag ctgtccttcc
      121 ccgagggctt caagtgggag agagtgatga acttcgagga cggcggcgtg gtgaccgtga
      181 cccaggactc ctccctgcag gacggcacct tcatctacca cgtgaagttc atcggcgtga
      241 acttcccctc cgacggcccc gtaatgcaga agaagactct gggctgggag ccctccaccg
      301 agagactgta ccccagagac ggcgtgctga agggcgagat ccacaaggcg ctgaagctga
      361 agggcggcgg ccactacctg gtggagttca agtcaatcta catggccaag aagcccgtga
      421 agctgcccgg ctactactac gtggactcca agctggacat cacctcccac aacgaggact
      481 acaccgtggt ggagcagtac gagagagccg aggccagaca ccacctgttc ttatagatcg
      541 ataagcttga tatcgaattc ctgcagcccg ggggatccac tagttctaga aactaagatt
      601 aatataatta tataaaaata ttatcttctt ttctttatat ctagtgttat gtaaaataaa
      661 ttgatgacta cggaaagctt ttttatattg tttctttttc attctgagcc acttaaattt
      721 cgtgaatgtt cttgtaaggg acggtagatt tacaagtgat acaacaaaaa gcaaggcgct
      781 ttttctaata aaaagaagaa aagcatttaa caattgaaca cctctatatc aacgaagaat
      841 attgctttgt ctctaaatcc ttgtaaaatg tgtacgatct ctatatgggt tactcataag
      901 tgtaccgaag actgcattga aagtttatgt tttttcactg gaggcgtcat tttcgcgttg
      961 agaagatgtt cttatccaaa tttcaactgt tatatagaag agcaaaaaat tgccaaaaaa
     1021 aacaacattt attcatttga aatataaaat ttgggcttct atattttaat attgcttttc
     1081 aattactgtt attaaatgta agtactgtgt ctatgaaaat atatgcaaat gctaagaaaa
     1141 atcctaaaaa tttgaatatg agatattcct cagtatttct ttttcatcct ttcttctgcg
     1201 gctctagccc tttgttctct catcaatctg cgtctctgtt catcggtcaa agaattcaaa
     1261 ttttgttgct gaattgaagg aatattggtt aactttttgc ctggacccct ctctgcatct
     1321 ttagcaaatc tttgtttttc cttggcggct tctaattcca atttaatcag tttttctgcc
     1381 ttttcattct ctattgcttt tttcttttca ttgatttcat caacttgttc tctgctcaat
     1441 ggctgaccga tatgagtttc gtcccaatta gtgaaagact ctccaagatt gtatcttgca
     1501 atttgaaacg cgtgtctcac gtcgtcgttt ctattcaaat cccacgcggc cgccaccgcg
     1561 gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     1621 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     1681 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     1741 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1801 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1861 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1921 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1981 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2041 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2101 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2161 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2221 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2281 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2341 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2401 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2461 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2521 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2581 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2641 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2701 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2761 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2821 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2881 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     2941 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3001 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3061 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3121 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3181 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3241 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3301 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3361 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3421 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3481 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3541 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3601 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3661 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3721 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcctt
     3781 ttcatcacgt gctataaaaa taattataat ttaaattttt taatataaat atataaatta
     3841 aaaatagaaa gtaaaaaaag aaattaaaga aaaaatagtt tttgttttcc gaagatgtaa
     3901 aagactctag ggggatcgcc aacaaatact accttttatc ttgctcttcc tgctctcagg
     3961 tattaatgcc gaattgtttc atcttgtctg tgtagaagac cacacacgaa aatcctgtga
     4021 ttttacattt tacttatcgt taatcgaatg tatatctatt taatctgctt ttcttgtcta
     4081 ataaatatat atgtaaagta cgctttttgt tgaaattttt taaacctttg tttatttttt
     4141 tttcttcatt ccgtaactct tctaccttct ttatttactt tctaaaatcc aaatacaaaa
     4201 cataaaaata aataaacaca gagtaaattc ccaaattatt ccatcattaa aagatacgag
     4261 gcgcgtgtaa gttacaggca agcgatccgt ctaagaaacc attattatca tgacattaac
     4321 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     4381 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     4441 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     4501 ctatgcggca tcagagcaga ttgtactgag agtgcaccat accacctttt caattcatca
     4561 tttttttttt attctttttt ttgatttcgg tttccttgaa atttttttga ttcggtaatc
     4621 tccgaacaga aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata
     4681 tgtagtgttg aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa
     4741 acctgcagga aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact
     4801 catcctagtc ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg
     4861 tgtgcttcat tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt
     4921 cccaaaattt gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc
     4981 acagttaagc cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaagacaga
     5041 aaatttgctg acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata
     5101 gcagaatggg cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt
     5161 ttgaagcagg cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa
     5221 ttgtcatgca agggctccct atctactgga gaatatacta agggtactgt tgacattgcg
     5281 aagagcgaca aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat
     5341 gaaggttacg attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagacgca
     5401 ttgggtcaac agtatagaac cgtggatgat gtggtctcta caggatctga cattattatt
     5461 gttggaagag gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga
     5521 aaagcaggct gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta
     5581 taagtaaatg catgtatact aaactcacaa attagagctt caatttaatt atatcagtta
     5641 ttaccctatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa
     5701 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     5761 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     5821 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     5881 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     5941 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     6001 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     6061 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     6121 cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gccattcagg ctgcgcaact
     6181 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     6241 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     6301 cgacggccag tgagcgcgcg taatacgact cactataggg cgaattgggt accgggcccc
     6361 ccctcgaggt cgacccaaga ataagggaac aagattttgg taatttccaa aaaatcaata
     6421 gcatgcagga cgttatgaag aagagatcta cgtatggtca tttcttcttc agattccctc
     6481 atggagaaag tgcggcagat gtatatgaca gagtcgccag tttccaagag actttattca
     6541 ggcacttcca tgataggcaa gagagaagac ccagagatgt tgttgtccta gttacacatg
     6601 gtatttattc cagagtattc ctgatgaaat ggtttagatg gacatacgaa gagtttgaat
     6661 cgtttaccaa tgttcctaac gggagcgtaa tggtgatgga actggacgaa tccatcaata
     6721 gatacgtcct gaggaccgtg ctacccaaat ggactgattg tgagggagac ctaactacat
     6781 agtgtttaaa gattacggat atttaactta cttagaataa tgccattttt ttgagttata
     6841 ataatcctac gttagtgtga gcgggattta aactgtgagg accttaatac attcagacac
     6901 ttctgcggta tcaccctact tattcccttc gagattatat ctaggaaccc atcaggttgg
     6961 tggaagatta cccgttctaa gacttttcag cttcctctat tgatgttaca cctggacacc
     7021 ccttttctgg catccagttt ttaatcttca gtggcatgtg agattctccg aaattaatta
     7081 aagcaatcac acaattctct cggataccac ctcggttgaa actgacaggt ggtttgttac
     7141 gcatgctaat gcaaaggagc ctatatacct ttggctcggc tgctgtaaca gggaatataa
     7201 agggcagcat aatttaggag tttagtgaac ttgcaacatt tactattttc ccttcttacg
     7261 taaatatttt tctttttaat tctaaatcaa tctttttcaa ttttttgttt gtattctttt
     7321 cttgcttaaa tctataacta caaaaaacac atacataaac taaaaatggc tagaactttc
     7381 tttgtcgaca tggtttccaa aggtgaagag aacaatatgg ctgtcatcaa gcccttcatg
     7441 agattcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     7501 ggcgagggca agccctacga gggcacccag accgccaagc tgcagg
//
