LOCUS       sequence-257193-        8280 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PADH1_sFUS_Fusion Red_PixD_TACT1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8280)
  AUTHORS   Zhao EM, Suek N, Wilson MZ, Dine E, Pannucci NL, Gitai Z, Avalos JL,
            Toettcher JE
  TITLE     Light-based control of metabolic flux through assembly of synthetic 
            organelles.
  JOURNAL   Nat Chem Biol. 2019 Jun;15(6):589-597. doi: 
            10.1038/s41589-019-0284-8. Epub 2019 May 13.
  PUBMED    31086330
REFERENCE   2  (bases 1 to 8280)
  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"; doi: 
            "10.1038/s41589-019-0284-8"; journalName: "Nat Chem Biol"; date: 
            "2019-06"; volume: "15"; issue: "6"; pages: "589-597"
COMMENT     Sequence Label: EZ-L767
FEATURES             Location/Qualifiers
     source          1..8280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        57..761
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1073..1768
                     /codon_start=1
                     /product="monomeric red fluorescent protein optimized for 
                     use as a fusion tag (Shemiakina et al., 2012)"
                     /label=FusionRed
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMPMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGACDMAL
                     KLVGGGHLICNLETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARY
                     STGGAGDGGK"
     primer_bind     complement(2254..2270)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2255..2271)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2814..2833)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2815..2833)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2840..2857)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2840..2856)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2849..2871)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2997..3452
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3084..3103)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3294..3315
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3808..3827)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3927..3949
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3987..4005)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4089..4193
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4194..4262,4263..5054)
                     /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(4412..4431)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5225..5813
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5714..5733
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5967..5984
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6101..6122
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6137..6142,6143..6160,6161..6167)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6175..6191
                     /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     6180..6202
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6199..6215
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6199..6215
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6236..6254
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    complement(6462..6495)
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            complement(6547..7467)
                     /label=bleMX6
                     /note="yeast selectable marker conferring phleomycin 
                     resistance"
     terminator      complement(6547..6743)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(6749..7123)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(7124..7467)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     protein_bind    complement(7522..7555)
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 taacacccct ccgcgctctt ttccgatttt tttctaaacc gtggaatatt tcggatatcc
       61 ttttgttgtt tccgggtgta caatatggac ttcctctttt ctggcaacca aacccataca
      121 tcgggattcc tataatacct tcgttggtct ccctaacatg taggtggcgg aggggagata
      181 tacaatagaa cagataccag acaagacata atgggctaaa caagactaca ccaattacac
      241 tgcctcattg atggtggtac ataacgaact aatactgtag ccctagactt gatagccatc
      301 atcatatcga agtttcacta ccctttttcc atttgccatc tattgaagta ataataggcg
      361 catgcaactt cttttctttt tttttctttt ctctctcccc cgttgttgtc tcaccatatc
      421 cgcaatgaca aaaaaatgat ggaagacact aaaggaaaaa attaacgaca aagacagcac
      481 caacagatgt cgttgttcca gagctgatga ggggtatctc gaagcacacg aaactttttc
      541 cttccttcat tcacgcacac tactctctaa tgagcaacgg tatacggcct tccttccagt
      601 tacttgaatt tgaaataaaa aaaagtttgc tgtcttgcta tcaagtataa atagacctgc
      661 aattattaat cttttgtttc ctcgtcattg ttctcgttcc ctttcttcct tgtttctttt
      721 tctgcacaat atttcaagct ataccaagca tacaatcaac tccattgcgg ccgctaaaat
      781 catggcctca aacgattata cccaacaagc aacccaaagc tatggggcct accccaccca
      841 gcccgggcag ggctattccc agcagagcag tcagccctac ggacagcaga gttacagtgg
      901 ttatagccag tccacggaca cttcaggcta tggccagagc agctattctt cttatggcca
      961 gagccagaac acaggctatg gaactcagtc aactccccag ggatatggct cgactggcgg
     1021 ctatggcagt agccagagct cccaatcgtc ttacgggcag cagggtggag gaatggtgag
     1081 cgagctgatt aaggagaaca tgcccatgaa gctgtacatg gagggcaccg tgaacaacca
     1141 ccacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag
     1201 aatcaaggtc gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctaccagctt
     1261 catgtacggc agcagaacct tcatcaagca ccctccgggc atccccgact tctttaagca
     1321 gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct
     1381 gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg tcaaggttag
     1441 aggggtgaac ttcccagcca acggccctgt gatgcagaag aaaacactcg gctgggaggc
     1501 ctccaccgag acgatgtacc ccgctgacgg cggcctggaa ggcgcatgtg acatggccct
     1561 gaagctcgtg ggcgggggcc acctgatctg caaccttgag accacataca gatccaagaa
     1621 acccgctacg aacctcaaga tgcccggcgt ctacaacgtg gaccacagac tggaaagaat
     1681 caaggaggcc gacgatgaga cctacgtcga gcagcacgag gtggctgtgg ccagatactc
     1741 tactggtggc gctggtgatg gaggtaaagc ggccacgcgt atgagtttgt accgtttgat
     1801 ttacagcagt cagggcattc ccaatttaca accccaggat ttaaaagata tcttagaatc
     1861 ttcccaaaga aataatccgg ccaatggcat taccggactg ctctgctaca gcaagccggc
     1921 ttttctgcag gtattggaag gagagtgtga acaggttaat gaaacctacc accggattgt
     1981 gcaagatgaa cgccaccaca gcccccagat cattgagtgt atgccgatca gaagaagaaa
     2041 cttcgaggtt tggtctatgc aagcgatcac ggtgaatgat ttgtcaacgg aacaggttaa
     2101 gactttggta ttgaaatatt ccggcttcac caccctacgg cccagtgcca tggacccgga
     2161 acaatgccta aactttttgc tagacattgc caagatttac gagttgtccg ataacttttt
     2221 cctcgacctc gaattcgata tctaaaagct tatcgatacc gtcgacctcg agtaatgatc
     2281 atctctgctt ttgtgcgcgt atgtttatgt atgtacctct ctctctattt ctatttttaa
     2341 accaccctct caataaaata aaaataataa agtattttta aggaaaagac gtgtttaagc
     2401 actgacttta tctacttttt gtacgttttc attgatataa tgtgttttgt ctctcccttt
     2461 tctacgaaaa tttcaaaaat tgaccaaaaa aaggaatata tatacgaaaa actattatat
     2521 ttatatatca tagtgttgat aaaaaatgtt tatccattgg accgtgtatc atatttatcg
     2581 ccggcgaata aattaaggcg cgccaacata tccggaggta ccataaaacg gaatgaggaa
     2641 taatcgtaat attagtatgt agaaatatag attccatttt gaggattcct atatcctcga
     2701 ggagaacttc tagtatattc tgtataccta atattatagc ctttatcaac aatggaatcc
     2761 caacaattat ctcaacattc acccatttct cagtttaaac ggccacgctg gccccctata
     2821 gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     2881 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     2941 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggacgc
     3001 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3061 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3121 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3181 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3241 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3301 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3361 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3421 gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat
     3481 ctgtgcggta tttcacaccg catagggtaa taactgatat aattaaattg aagctctaat
     3541 ttgtgagttt agtattagga tctctaccct tggcgaaaag tcctctgcca acaatgatga
     3601 tatctgatcc accacttaca acttcgtcga gggttctgta ctgctgaccc aatatgcgta
     3661 tatataccaa tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg
     3721 aatcaaaaaa atttcaagga aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat
     3781 tgaaaaggtg gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3841 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3901 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3961 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4021 catgataata atggtttctt agacgtcacg cgccgaggac gtcaggtggc acttttcggg
     4081 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4141 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4201 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4261 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4321 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4381 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4441 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4501 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4561 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4621 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4681 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4741 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4801 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4861 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4921 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4981 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5041 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5101 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5161 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5221 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5281 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5341 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5401 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5461 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5521 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5581 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5641 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5701 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5761 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5821 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5881 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     5941 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     6001 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     6061 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     6121 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     6181 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ctcggaatta
     6241 accctcacta aagggaacaa aagctgggcc ggccatttaa atgtttaaac aacctgttgg
     6301 aatagaaatc aactatcatc tactaactag tatttacatt actagtatat tatcatatac
     6361 ggtgttagaa gatgacgcaa atgatgagaa atagtcatct aaattagtgg aagctgaaac
     6421 gcaaggattg ataatgtaat aggatcaatg aatataaaca ttaccgttcg tatagcatac
     6481 attatacgaa gttatattaa gggttctcga gagctcggga tggcggcgtt agtatcgaat
     6541 cgacagcagt atagcgacca gcattcacat acgattgacg catgatatta ctttctgcgc
     6601 acttaacttc gcatctgggc agatgatgtc gaggcgaaaa aaatataaat cacgctaaca
     6661 tttgattaaa atagaacaac tacaatataa aaaaactata caaatgtcaa gttcttgaaa
     6721 acaagaatct ttttattgtc agtactgatc agtcctgctc ctcggccacg aagtgcacgc
     6781 agttgccggc cgggtcgcgc agggcgaact cccgccccca cggctgctcg ccgatctcgg
     6841 tcatggccgg cccggaggcg tcccggaagt tcgtggacac gacctccgac cactcggcgt
     6901 acagctcgtc caggccgcgc acccacaccc aggccagggt gttgtccggc accacctggt
     6961 cctggaccgc gctgatgaac agggtcacgt cgtcccggac cacaccggcg aagtcgtcct
     7021 ccacgaagtc ccgggagaac ccgagccggt cggtccagaa ctcgaccgct ccggcgacgt
     7081 cgcgcgcggt gagcaccgga acggcgctgg tcaacttggc catggttgtt tatgttcgga
     7141 tgtgatgtga gaactgtatc ctagcaagat tttaaaagga agtatatgaa agaagaacct
     7201 cagtggcaaa tcctaacctt ttatatttct ctacaggggc gcggcgtggg gacaattcaa
     7261 cgcgtctgtg aggggagcgt ttccctgctc gcaggtctgc agcgaggagc cgtaattttt
     7321 gcttcgcgcc gtgcggccat caaaatgtat ggatgcaaat gattatacat ggggatgtat
     7381 gggctaaatg tacgggcgac agtcacatca tgcccctgag ctgcgcacgt caagactgtc
     7441 aaggagggta ttctgggcct ccatgtcgct ggccgggtga cccggcgggg acgaggcaag
     7501 ctaaacagat ctctagacct aataacttcg tatagcatac attatacgaa cggtacccgg
     7561 gaaacagctg cgccgggaaa cagctggccg ggatcgaaga aatgatggta aatgaaatag
     7621 gaaatcaagg agcatgaagg caaaagacaa atataagggt cgaacgaaaa ataaagtgaa
     7681 aagtgttgat atgatgtatt tggctttgcg gcgccgaaaa aacgagttta cgcaattgca
     7741 caatcatgct gactctgtgg cggacccgcg ctcttgccgg cccggcgata acgctgggcg
     7801 tgaggctgtg cccggcggag ttttttgcgc ctgcattttc caaggtttac cctgcgctaa
     7861 ggggcgagat tggagaagca ataagaatgc cggttggggt tgcgatgatg acgaccacga
     7921 caactggtgt cattatttaa gttgccgaaa gaacctgagt gcatttgcaa catgagtata
     7981 ctagaagaat gagccaagac ttgcgagacg cgagtttgcc ggtggtgcga acaatagagc
     8041 gaccatgacc ttgaaggtga gacgcgcata accgctagag tactttgaag aggaaacagc
     8101 aatagggttg ctaccagtat aaatagacag gtacatacaa cactggaaat ggttgtctgt
     8161 ttgagtacgc tttcaattca tttgggtgtg cactttatta tgttacaata tggaagggaa
     8221 ctttacactt ctcctatgca catatattaa ttaaagtcca atgctagtag agaagggggg
//
