LOCUS       sequence-253019-        8281 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 8281)
  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 8281)
  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..8281
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    13..34
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(49..54,55..72,73..79)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    87..103
                     /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     92..114
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     111..127
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     111..127
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        148..166
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    complement(374..407)
                     /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(459..1380)
                     /label=bleMX6
                     /note="yeast selectable marker conferring phleomycin 
                     resistance"
     terminator      complement(459..656)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(662..1036)
                     /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(1037..1380)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     protein_bind    complement(1435..1468)
                     /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)."
     promoter        2250..2954
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             3266..3961
                     /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(4447..4463)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4448..4464)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5007..5026)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5008..5026)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5033..5050)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5033..5049)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5042..5064)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5190..5645
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5277..5296)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5487..5508
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6001..6020)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6120..6142
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6180..6198)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6282..6386
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6387..6455,6456..7247)
                     /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(6605..6624)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7418..8006
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7907..7926
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8160..8177
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
       61 gcttccggct cctatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
      121 ctatgaccat gattacgcca agctcggaat taaccctcac taaagggaac aaaagctggg
      181 ccggccattt aaatgtttaa acaacctgtt ggaatagaaa tcaactatca tctactaact
      241 agtatttaca ttactagtat attatcatat acggtgttag aagatgacgc aaatgatgag
      301 aaatagtcat ctaaattagt ggaagctgaa acgcaaggat tgataatgta ataggatcaa
      361 tgaatataaa cattaccgtt cgtatagcat acattatacg aagttatatt aagggttctc
      421 gagagctcgg gatggcggcg ttagtatcga atcgacagca gtatagcgac cagcattcac
      481 atacgattga cgcatgatat tactttctgc gcacttaact tcgcatctgg gcagatgatg
      541 tcgaggcgaa aaaaaatata aatcacgcta acatttgatt aaaatagaac aactacaata
      601 taaaaaaact atacaaatga caagttcttg aaaacaagaa tctttttatt gtcagtactg
      661 atcagtcctg ctcctcggcc acgaagtgca cgcagttgcc ggccgggtcg cgcagggcga
      721 actcccgccc ccacggctgc tcgccgatct cggtcatggc cggcccggag gcgtcccgga
      781 agttcgtgga cacgacctcc gaccactcgg cgtacagctc gtccaggccg cgcacccaca
      841 cccaggccag ggtgttgtcc ggcaccacct ggtcctggac cgcgctgatg aacagggtca
      901 cgtcgtcccg gaccacaccg gcgaagtcgt cctccacgaa gtcccgggag aacccgagcc
      961 ggtcggtcca gaactcgacc gctccggcga cgtcgcgcgc ggtgagcacc ggaacggcgc
     1021 tggtcaactt ggccatggtt gtttatgttc ggatgtgatg tgagaactgt atcctagcaa
     1081 gattttaaaa ggaagtatat gaaagaagaa cctcagtggc aaatcctaac cttttatatt
     1141 tctctacagg ggcgcggcgt ggggacaatt caacgcgtct gtgaggggag cgtttccctg
     1201 ctcgcaggtc tgcagcgagg agccgtaatt tttgcttcgc gccgtgcggc catcaaaatg
     1261 tatggatgca aatgattata catggggatg tatgggctaa atgtacgggc gacagtcaca
     1321 tcatgcccct gagctgcgca cgtcaagact gtcaaggagg gtattctggg cctccatgtc
     1381 gctggccggg tgacccggcg gggacgaggc aagctaaaca gatctctaga cctaataact
     1441 tcgtatagca tacattatac gaacggtacc cgggaaacag ctgcgccggg aaacagctgg
     1501 ccgggatcga agaaatgatg gtaaatgaaa taggaaatca aggagcatga aggcaaaaga
     1561 caaatataag ggtcgaacga aaaataaagt gaaaagtgtt gatatgatgt atttggcttt
     1621 gcggcgccga aaaaacgagt ttacgcaatt gcacaatcat gctgactctg tggcggaccc
     1681 gcgctcttgc cggcccggcg ataacgctgg gcgtgaggct gtgcccggcg gagttttttg
     1741 cgcctgcatt ttccaaggtt taccctgcgc taaggggcga gattggagaa gcaataagaa
     1801 tgccggttgg ggttgcgatg atgacgacca cgacaactgg tgtcattatt taagttgccg
     1861 aaagaacctg agtgcatttg caacatgagt atactagaag aatgagccaa gacttgcgag
     1921 acgcgagttt gccggtggtg cgaacaatag agcgaccatg accttgaagg tgagacgcgc
     1981 ataaccgcta gagtactttg aagaggaaac agcaataggg ttgctaccag tataaataga
     2041 caggtacata caacactgga aatggttgtc tgtttgagta cgctttcaat tcatttgggt
     2101 gtgcacttta ttatgttaca atatggaagg gaactttaca cttctcctat gcacatatat
     2161 taattaaagt ccaatgctag tagagaaggg gggtaacacc cctccgcgct cttttccgat
     2221 ttttttctaa accgtggaat atttcggata tccttttgtt gtttccgggt gtacaatatg
     2281 gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata ccttcgttgg
     2341 tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac cagacaagac
     2401 ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg tacataacga
     2461 actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca ctaccctttt
     2521 tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct ttttttttct
     2581 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat gatggaagac
     2641 actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt ccagagctga
     2701 tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca cactactctc
     2761 taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata aaaaaaagtt
     2821 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca
     2881 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa
     2941 gcatacaatc aactccattg cggccgctaa aatcatggcc tcaaacgatt atacccaaca
     3001 agcaacccaa agctatgggg cctaccccac ccagcccggg cagggctatt cccagcagag
     3061 cagtcagccc tacggacagc agagttacag tggttatagc cagtccacgg acacttcagg
     3121 ctatggccag agcagctatt cttcttatgg ccagagccag aacacaggct atggaactca
     3181 gtcaactccc cagggatatg gctcgactgg cggctatggc agtagccaga gctcccaatc
     3241 gtcttacggg cagcagggtg gaggaatggt gagcgagctg attaaggaga acatgcccat
     3301 gaagctgtac atggagggca ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga
     3361 aggcaagccc tacgagggca cccagaccat gagaatcaag gtcgtcgagg gcggccctct
     3421 ccccttcgcc ttcgacatcc tggctaccag cttcatgtac ggcagcagaa ccttcatcaa
     3481 gcaccctccg ggcatccccg acttctttaa gcagtccttc cctgagggct tcacatggga
     3541 gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca
     3601 ggacggctgc ctcatctaca acgtcaaggt tagaggggtg aacttcccag ccaacggccc
     3661 tgtgatgcag aagaaaacac tcggctggga ggcctccacc gagacgatgt accccgctga
     3721 cggcggcctg gaaggcgcat gtgacatggc cctgaagctc gtgggcgggg gccacctgat
     3781 ctgcaacctt gagaccacat acagatccaa gaaacccgct acgaacctca agatgcccgg
     3841 cgtctacaac gtggaccaca gactggaaag aatcaaggag gccgacgatg agacctacgt
     3901 cgagcagcac gaggtggctg tggccagata ctctactggt ggcgctggtg atggaggtaa
     3961 agcggccacg cgtatgagtt tgtaccgttt gatttacagc agtcagggca ttcccaattt
     4021 acaaccccag gatttaaaag atatcttaga atcttcccaa agaaataatc cggccaatgg
     4081 cattaccgga ctgctctgct acagcaagcc ggcttttctg caggtattgg aaggagagtg
     4141 tgaacaggtt aatgaaacct accaccggat tgtgcaagat gaacgccacc acagccccca
     4201 gatcattgag tgtatgccga tcagaagaag aaacttcgag gtttggtcta tgcaagcgat
     4261 cacggtgaat gatttgtcaa cggaacaggt taagactttg gtattgaaat attccggctt
     4321 caccacccta cggcccagtg ccatggaccc ggaacaatgc ctaaactttt tgctagacat
     4381 tgccaagatt tacgagttgt ccgataactt tttcctcgac ctcgaattcg atatctaaaa
     4441 gcttatcgat accgtcgacc tcgagtaatg atcatctctg cttttgtgcg cgtatgttta
     4501 tgtatgtacc tctctctcta tttctatttt taaaccaccc tctcaataaa ataaaaataa
     4561 taaagtattt ttaaggaaaa gacgtgttta agcactgact ttatctactt tttgtacgtt
     4621 ttcattgata taatgtgttt tgtctctccc ttttctacga aaatttcaaa aattgaccaa
     4681 aaaaaggaat atatatacga aaaactatta tatttatata tcatagtgtt gataaaaaat
     4741 gtttatccat tggaccgtgt atcatattta tcgccggcga ataaattaag gcgcgccaac
     4801 atatccggag gtaccataaa acggaatgag gaataatcgt aatattagta tgtagaaata
     4861 tagattccat tttgaggatt cctatatcct cgaggagaac ttctagtata ttctgtatac
     4921 ctaatattat agcctttatc aacaatggaa tcccaacaat tatctcaaca ttcacccatt
     4981 tctcagttta aacggccacg ctggccccct atagtgagtc gtattacaat tcactggccg
     5041 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     5101 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     5161 aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc
     5221 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     5281 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     5341 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     5401 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     5461 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     5521 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     5581 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     5641 aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataggg
     5701 taataactga tataattaaa ttgaagctct aatttgtgag tttagtatta ggatctctac
     5761 ccttggcgaa aagtcctctg ccaacaatga tgatatctga tccaccactt acaacttcgt
     5821 cgagggttct gtactgctga cccaatatgc gtatatatac caatctaagt ctgtgctcct
     5881 tccttcgttc ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa
     5941 atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag gtggtatggt gcactctcag
     6001 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     6061 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     6121 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     6181 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     6241 acgcgccgag gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     6301 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     6361 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     6421 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     6481 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     6541 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     6601 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     6661 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     6721 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6781 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     6841 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6901 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     6961 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     7021 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     7081 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     7141 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     7201 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     7261 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     7321 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     7381 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     7441 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     7501 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     7561 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     7621 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     7681 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     7741 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     7801 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     7861 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     7921 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     7981 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     8041 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     8101 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     8161 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     8221 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     8281 a
//
