LOCUS       sequence-423112-        9010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GFP-Cps1 under GAL promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9010)
  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 9010)
  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: BS-TRP1-PGAL-GFP-Cps1
FEATURES             Location/Qualifiers
     source          1..9010
                     /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"
     promoter        1991..2432
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1991..2108
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2403..2426
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2444..3157
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGIDELYK"
     primer_bind     complement(5197..5213)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5197..5213)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(6240..6259)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6241..6259)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6266..6283)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6266..6282)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6275..6297)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6424..6879
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6511..6530)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6721..6742
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6905..7009
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7010..7078,7079..7870)
                     /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(7228..7247)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8041..8629
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8530..8549
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8783..8800
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8917..8938
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8953..8958,8959..8976,8977..8983)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8991..9007
                     /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(8996..9010,1..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 agcttggcat taccaccata tacatatcca
     1861 tatacatatc catatctaat cttacttata tgttgtggaa atgtaaagag ccccattatc
     1921 ttagcctaaa aaaaccttct ctttggaact ttcagtaata cgcttaactg ctcattgcta
     1981 tattgaagta cggattagaa gccgccgagc gggtgacagc cctccgaagg aagactctcc
     2041 tccgtgcgtc ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca
     2101 ctgctccgaa caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt
     2161 ggcagtaacc tggccccaca aaccttcaaa tgaacgaatc aaattaacaa ccataggatg
     2221 ataatgcgat tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt
     2281 ttgatctatt aacagatata taaatgcaaa aactgcataa ccactttaac taatactttc
     2341 aacattttcg gtttgtatta cttcttattc aaatgtaata aaagtatcaa caaaaaattg
     2401 ttaatatacc tctatacttt aacgtcaagg agaaaaaact ataatgtcta aaggtgaaga
     2461 attattcact ggtgttgtcc caattttggt tgaattagat ggtgatgtta atggtcacaa
     2521 attttctgtc tccggtgaag gtgaaggtga tgctacttac ggtaaattga ccttaaaatt
     2581 tatttgtact actggtaaat tgccagttcc atggccaacc ttagtcacta ctttaactta
     2641 tggtgttcaa tgtttttcta gatacccaga tcatatgaaa caacatgact ttttcaagtc
     2701 tgccatgcca gaaggttatg ttcaagaaag aactattttt ttcaaagatg acggtaacta
     2761 caagaccaga gctgaagtca agtttgaagg tgatacctta gttaatagaa tcgaattaaa
     2821 aggtattgat tttaaagaag atggtaacat tttaggtcac aaattggaat acaactataa
     2881 ctctcacaat gtttacatca tggctgacaa acaaaagaat ggtatcaaag ttaacttcaa
     2941 aattagacac aacattgaag atggttctgt tcaattagct gaccattatc aacaaaatac
     3001 tccaattggt gatggtccag tcttgttacc agacaaccat tacttatcca ctcaatctgc
     3061 cttatccaaa gatccaaacg aaaagagaga ccacatggtc ttgttagaat ttgttactgc
     3121 tgctggtatt acccatggta ttgatgaatt gtacaaagga gccggaacaa gtgacatgat
     3181 cgccttacca gtagagaagg cccctagaaa gtccctatgg caaaggcaca gagcctttat
     3241 tagtggaata gttgccctta ttatcatcgg caccttcttc ctcacttcgg gtctccaccc
     3301 agcaccacct catgaggcaa agcgtccaca ccatggaaaa ggtcccatgc actcacccaa
     3361 atgtgagaag attgaaccat taagtccatc attcaaacat tccgtcgaca caattcttca
     3421 tgaccctgcc tttagaaaca gctccattga gaaactgtcc aatgctgtta gaatccccac
     3481 tgtagtccaa gacaaaaacc ccaaccccgc agatgatccg gatttctata agcattttta
     3541 tgaactacac gactattttg agaagacttt ccctaatatt cacaagcatt tgaaattgga
     3601 gaaagtcaat gagctgggtc ttctatacac atgggaaggt tctgatcctg atctaaaacc
     3661 attattgtta atggcccatc aagatgttgt acctgtaaac aacgaaactt tatcatcctg
     3721 gaagttccct ccattttctg gtcactatga tccagaaaca gattttgttt gggggcgtgg
     3781 ttctaacgat tgtaagaact tgttaattgc cgagtttgaa gctatcgaac aactgttgat
     3841 agatggattc aagcccaaca gaactattgt tatgtcgctt ggttttgatg aagaagcaag
     3901 cggcaccctc ggtgctgcca gcttagcctc atttcttcac gaaagatatg gtgatgatgg
     3961 tatttacagt atcattgacg agggtgaagg tatcatggaa gtcgacaagg atgtctttgt
     4021 tgccactcca atcaacgctg aaaaaggcta tgtcgacttc gaagtcagta ttctaggcca
     4081 tggtggtcat tcctctgtcc cacctgatca taccacaatc ggtatcgctt cagaactgat
     4141 tactgaattt gaagccaacc catttgacta cgaatttgag tttgacaatc caatctatgg
     4201 attgttgaca tgtgctgctg aacattctaa atctttaagt aaggatgtga agaagacaat
     4261 tttgggcgca ccattctgtc ctagaaggaa ggacaagctt gttgagtaca tttccaacca
     4321 atcacatttg cgcagcttaa taagaacaac acaagctgtt gatatcatca atggtggtgt
     4381 taaagctaat gctctgcccg aaactaccag attcttgatc aatcacagaa ttaatttaca
     4441 ttcttctgtg gctgaagtct ttgaaagaaa catagaatat gcgaaaaaga ttgctgagaa
     4501 gtatggctat ggtttatcta agaacggtga cgattacatt atccctgaaa ccgagttagg
     4561 tcacattgac attactctct tgagagaatt ggaaccagca ccactttcgc caagttcagg
     4621 ccctgtttgg gacattttgg caggtactat tcaagatgtt tttgaaaacg gtgttctaca
     4681 aaacaacgaa gagttctatg tgactactgg tttattctct ggtaacaccg atactaaata
     4741 ctactggaat ttgtccaaga acatttatag gtttgttggc tctatcattg atattgattt
     4801 actgaagaca ttgcattcgg ttaatgaaca cgtggatgtc ccaggtcatt tatctgccat
     4861 tgcctttgtt tacgagtata tcgttaatgt taacgaatac gcttaagcgc aatgattgaa
     4921 tagtcaaaga tttttttttt ttaatttttt ttttttaatt tttttttttt ttcatagaac
     4981 tttttattta aataaatcac gtctatatat gtatcagtat aacgtaaaaa aaaaaacacc
     5041 gtcagttaaa caaaacataa ataaaaaaaa aaagaagtgt caaatcaagt gtcaaatcaa
     5101 aatttaaatg atgatgataa tatgaccata caatgactta aatcaaatca aaaccacttg
     5161 cgtaaaggct tgatatcgaa ttcctgcagc ccgggggatc cactagttct agagcggccg
     5221 ccaccgcggt gggttattac tgagtagtat ttatttaagt attgtttgtg cacttgcctg
     5281 caggcctttt gaaaagcaag cataaaagat ctaaacataa aatctgtaaa ataacaagat
     5341 gtaaagataa tgctaaatca tttggctttt tgattgattg tacaggaaaa tatacatcgc
     5401 agggggttga cttttaccat ttcaccgcaa tggaatcaaa cttgttgaag agaatgttca
     5461 caggcgcata cgctacaatg acccgattct tgctagcctt ttctcggtct tgcaaacaac
     5521 cgccggcagc ttagtatata aatacacatg tacatacctc tctccgtatc ctcgtaatca
     5581 ttttcttgta tttatcgtct tttcgctgta aaaactttat cacacttatc tcaaatacac
     5641 ttattaaccg cttttactat tatcttctac gctgacagta atatcaaaca gtgacacata
     5701 ttaaacacag tggtttcttt gcataaacac catcagcctc aagtcgtcaa gtaaagattt
     5761 cgtgttcatg cagatagata acaatctata tgttgataat tagcgttgcc tcatcaatgc
     5821 gagatccgtt taaccggacc ctagtgcact taccccacgt tcggtccact gtgtgccgaa
     5881 catgctcctt cactatttta acatgtggaa ttcttgaaag aatgaaatcg ccatgccaag
     5941 ccatcacacg gtcttttatg caattgattg accgcctgca acacataggc agtaaaattt
     6001 ttactgaaac gtatataatc atcataagcg acaagtgagg caacaccttt gttaccacat
     6061 tgacaacccc aggtattcat acttcctatt agcggaatca ggagtgcaaa aagagaaaat
     6121 aaaagtaaaa aggtagggca acacatagta tgaatacaaa cgttccaata ttcagttctc
     6181 cggtcagaga tttaccaagg tctttcgaac aaaaacattt agcggttgta gacaattcgc
     6241 cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     6301 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     6361 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     6421 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     6481 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     6541 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     6601 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     6661 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     6721 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     6781 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     6841 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     6901 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     6961 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     7021 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     7081 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     7141 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     7201 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     7261 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     7321 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     7381 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     7441 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     7501 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     7561 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     7621 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     7681 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     7741 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     7801 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     7861 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     7921 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     7981 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     8041 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     8101 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     8161 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     8221 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     8281 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     8341 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     8401 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     8461 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     8521 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     8581 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     8641 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     8701 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     8761 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     8821 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     8881 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     8941 ggcaccccag gctttacact ttatgctccc ggctcgtatg ttgtgtggaa ttgtgagcgg
     9001 ataacaattt
//
