LOCUS       sequence-377457-        7633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express C.elegans LACE-1 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7633)
  AUTHORS   Germany EM, Zahayko N, Huebsch ML, Fox JL, Prahlad V, Khalimonchuk O
  TITLE     The AAA ATPase Afg1 preserves mitochondrial fidelity and cellular 
            health by maintaining mitochondrial matrix proteostasis.
  JOURNAL   J Cell Sci. 2018 Nov 21;131(22). pii: jcs.219956. doi: 
            10.1242/jcs.219956.
  PUBMED    30301782
REFERENCE   2  (bases 1 to 7633)
  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: "J Cell 
            Sci."; date: "2018-11-21"; volume: "131(22). pii: jcs.219956. doi"; 
            pages: " 10.1242/jcs.219956"
COMMENT     Sequence Label: pRS426-CeLACE-1
FEATURES             Location/Qualifiers
     source          1..7633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        194..409
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             410..1213
                     /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(1344..1799)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1481..1502)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1693..1712
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1925..1947
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1939..1956
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1940..1956
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1966..1985
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1966..1984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2003..2250)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2216..2234
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(2257..2274)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3614..3630)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3614..3630)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(3631..3980)
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     complement(4031..4051)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4031..4049)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4070..4086)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4070..4086)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4083..4105)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4094..4110
                     /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(4118..4124,4125..4142,4143..4148))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4163..4184
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4301..4318)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4472..5060)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4552..4571)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5231..6022,6023..6091))
                     /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     5854..5873
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6092..6196)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6223..7565
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(7186..7233)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     7606..7624
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc
      241 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
      301 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
      361 cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaatca tgtcgaaagc
      421 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
      481 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
      541 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
      601 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
      661 caatttttta ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca
      721 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
      781 ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc
      841 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata
      901 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc
      961 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
     1021 gggtttagat gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt
     1081 ctctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
     1141 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
     1201 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
     1261 agcttcaatt taattatatc agttattacc ctatgcggtg tgaaataccg cacagatgcg
     1321 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     1381 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     1441 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     1501 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     1561 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     1621 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     1681 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     1741 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     1801 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     1861 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     1921 ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta
     1981 tagggcgaat tgggtaccgg ccgcaaatta aagccttcga gcgtcccaaa accttctcaa
     2041 gcaaggtttt cagtataatg ttacatgcgt acacgcgtct gtacagaaaa aaaagaaaaa
     2101 tttgaaatat aaataacgtt cttaatacta acataactat aaaaaaataa atagggacct
     2161 agacttcagg ttgtctaact ccttcctttt cggttagagc ggatgtgggg ggagggcgtg
     2221 aatgtaagcg tgacataact aattacatga ctcgagatga tgatgatgat gatgaataga
     2281 atcttcagta gcattataag gtcttcgctg tctcctatat ttttctgttt gcatctcata
     2341 caatctggat actgtcctac tgtaggcgaa tgcttcttca tctccactga acacatttgc
     2401 gctcattcct tcatgatcag atttgatacc cagatcatcc atgagcattc gttttgagtc
     2461 acttagagcg tcgtgagaag tattgtgtcc ttcaaattga aagagctcat ctaaaggagc
     2521 agctgctcca attacaacac ggaccttctg atcataaaat gtgtcaatca ttgtgatgaa
     2581 tcgtctcata gcattccaca tatcttggtt cataattgga atatttctta caataactgt
     2641 atgaaatact cgtgcataaa caagataatc aactgctccc ttcgctgtca tgcagagctc
     2701 tttgaaatcc acatcagcaa ctccaccaca acatttctcg acgatcactc gacgtcccaa
     2761 gatttcgaga gttttcgaac gtacagtatc attctcatta gctgcagatt gtttgaaaac
     2821 tatgtcacat tgtgtattag aatcttctga actgaaataa acatgatttc cgtctccagc
     2881 tgcactgcga cggtaatcca ttccagaatc caaagcaaga gatgcacatt tatcttctaa
     2941 aatagtgatg aaaggcatga actgatgtcg ctgaagacca tttttgtaaa gttctgaagg
     3001 tgctcgattt gaagttgcaa ccattacaag accacgatca aataacattg agaagaatcg
     3061 ttttagaatc attgcgtcgg caatatctgt gacctgaaac tcatcaaaac acaataaatt
     3121 cgtcgtttcc ataatttcat cgacaataac tggtacagga tcaaattttc cacgagcttt
     3181 attactctgc attttgagtt cgtgcattct tttatgtaca ttttgcataa aatcattaaa
     3241 atgaacccgt cgttttttat ctatcggaca attttcaaaa aataaatcca tgagcattgt
     3301 tttcccgcat ccaactgaac catacaagta aattccacgt ggtgatataa tttttggagt
     3361 atctacttta gaattttgaa acattttcca gaatcttgaa cttgattttt ctgatatatt
     3421 agacttgttc gttggttgat atgattcgat ttctttccgc aatcgttcga aatcaacaat
     3481 catttttctc tgataatcgt cttctttaag tgtcccttcg tttacttttt tcgagtacgc
     3541 atcgcttaat gtgactaaaa accgttgaac ttgcagatat tgtaattttg agcgaattgg
     3601 ggagagcatc gcggatccac tagttctaga gtatggatgg gggtaataga attgtatcta
     3661 tgtatctgac gaccctgtat tacacgaagg aaatagtaga tgaaaagaca agagagcaag
     3721 aaaaaggaaa aacttccttt ctaacagacg ctttacttaa ccttatatat atcttatttt
     3781 tttcatcgag cgtgttcaat tggacaaggt gccacctttt cgacacttcg gttattatgt
     3841 tacacagttt tcatgaggat ggcgccttga ctaacttaat tagtcatttg ccaaccacca
     3901 cagttcccca atatcgacag cttcacgtgc catttgccat tttgcgatca tgtgacctca
     3961 agagaaaaag caaaaaataa tgagagctaa gggattcgaa cccttgcatc cggagctcca
     4021 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     4081 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4141 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4201 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4261 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4321 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4381 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4441 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4501 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4561 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4621 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4681 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4741 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4801 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4861 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4921 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4981 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5041 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5101 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5161 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5221 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5281 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5341 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5401 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5461 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5521 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5581 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5641 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5701 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5761 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5821 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5881 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5941 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6001 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6061 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6121 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6181 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgaacgaagc atctgtgctt
     6241 cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     6301 tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt
     6361 gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc
     6421 tgagctgcat ttttacagaa cggaaatgca acgcgagagc gctattttac caacaaagaa
     6481 tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag
     6541 catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt
     6601 ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca
     6661 taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat
     6721 tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt
     6781 gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc
     6841 ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga
     6901 ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa
     6961 acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt
     7021 aggttatata gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt
     7081 gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt
     7141 ttgaaagtgc gtcttcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt
     7201 tctagagaat aggaacttcg gaataggaac ttcaaagcgt ttccgaaaac gagcgcttcc
     7261 gaaaatgcaa cgcgagctgc gcacatacag ctcactgttc acgtcgcacc tatatctgcg
     7321 tgttgcctgt atatatatat acatgagaag aacggcatag tgcgtgttta tgcttaaatg
     7381 cgtacttata tgcgtctatt tatgtaggat gaaaggtagt ctagtacctc ctgtgatatt
     7441 atcccattcc atgcggggta tcgtatgctt ccttcagcac taccctttag ctgttctata
     7501 tgctgccact cctcaattgg attagtctca tccttcaatg ctatcatttc ctttgatatt
     7561 ggatcatact aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     7621 aggccctttc gtc
//
