LOCUS       sequence-387294-        7395 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of FRET_control  in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7395)
  AUTHORS   Xue L, Schnacke P, Frei MS, Koch B, Hiblot J, Wombacher R, Fabritz 
            S, Johnsson K
  TITLE     Probing coenzyme A homeostasis with semisynthetic biosensors.
  JOURNAL   Nat Chem Biol. 2022 Oct 31. doi: 10.1038/s41589-022-01172-7.
  PUBMED    36316571
REFERENCE   2  (bases 1 to 7395)
  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: "Nat Chem 
            Biol. 2022 Oct 31. doi: 10.1038/s41589-022-01172-7."
COMMENT     Sequence Label: pCDNA5/FRT/TO_COX8-Halo-PanK
FEATURES             Location/Qualifiers
     source          1..7395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(16..33)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(187..775)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(267..286)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(946..1737,1738..1806))
                     /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     1569..1588
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1807..1911)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1986..2005)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2177..2556
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2557..2760
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2711..2731
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2762..2780
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2783..2801
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2811..2835
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2938..3012
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             3025..3096
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     CDS             3115..3186
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     CDS             3199..3270
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     CDS             3301..4185
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     primer_bind     complement(5289..5306)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5295..5519
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5652..5671)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5803..5850
                     /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)."
     CDS             5858..6880
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             5858..6880
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    7010..7131
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7047..7066)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7101..7120
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7180..7196)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7180..7196)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7193..7215)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7204..7220
                     /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(7228..7234,7235..7252,7253..7258))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7273..7294
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
       61 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
      121 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      181 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      241 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      301 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      361 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      421 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      481 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      541 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      601 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
      661 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
      721 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
      781 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
      841 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
      901 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
      961 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1021 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1081 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1141 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1201 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1261 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1321 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1381 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1441 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1501 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     1561 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     1621 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     1681 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     1741 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     1801 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     1861 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     1921 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     1981 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     2041 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     2101 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
     2161 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
     2221 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     2281 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     2341 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     2401 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     2461 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     2521 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     2581 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     2641 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     2701 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     2761 ctccctatca gtgatagaga tctccctatc agtgatagag atcgtcgacg agctcgttta
     2821 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
     2881 cgggaccgat ccagcctccg gactctagcg tttaaactta agcttggtac cgccaccatg
     2941 tctgtgctga cacctctgct gctgagaggc ctgacaggct ctgctagaag gctgcctgtg
     3001 cctagagcca agatccactc tctgagcgtg ctgaccccac tgctccttag gggactgact
     3061 ggatctgctc ggagactgcc agtgcctcgg gccaaaattc atagcctcgg cggatccgtc
     3121 ctgactccac tccttctgag aggactcacc ggcagtgcta gacgcctgcc agttccaagg
     3181 gctaagatcc acagtctgtc agtgctcact cccctgttgc tcagagggct gactggaagt
     3241 gcaagacgac tccccgtgcc acgcgccaag attcattctc ttggaggctc tggcggcagc
     3301 gagatcggaa ccggctttcc tttcgaccct cactacgtgg aagtgctggg cgagagaatg
     3361 cactatgtgg acgtgggccc cagagatgga acccctgtgc tgtttctgca cggcaaccct
     3421 accagcagct acgtgtggcg gaacatcatc cctcacgtgg cccctacaca cagatgtatc
     3481 gcccctgacc tgatcggcat gggcaagagc gataagcccg acctgggcta cttcttcgac
     3541 gaccacgtgc ggttcatgga cgcctttatt gaggccctcg gcctggaaga ggtggtgctg
     3601 gttattcacg attggggcag cgccctggga ttccactggg ccaagagaaa ccccgagaga
     3661 gtgaagggaa tcgccttcat ggaattcatc aggcccattc ctacctggga cgagtggcct
     3721 gagttcgcca gagagacatt ccaggccttc agaaccaccg acgtgggcag aaagctgatc
     3781 atcgaccaga acgtgttcat cgagggcacc ctgcctatgg gagtcgttag acctctgacc
     3841 gaggtggaaa tggaccacta cagagagccc tttctgaacc ccgtggaccg ggaacctctt
     3901 tggagattcc ccaacgagct gcccattgcc ggcgagcctg ctaatattgt ggccctggtg
     3961 gaagagtaca tggactggct gcatcagagc cccgtgccta agctgctgtt ttggggaaca
     4021 cccggcgtgc tgattcctcc tgctgaagct gccagactgg ccaagagcct gcctaattgc
     4081 aaggccgtgg atatcggccc tggcctgaat ctgctgcaag aggacaaccc cgatctgatc
     4141 ggctccgaga ttgccagatg gctgagcacc ctggaaatca gcggaggatc tctggctgaa
     4201 gccgccgcta aagaagccgc tgcaaaagag gctgccgcca aagaggcagc agcaaaagaa
     4261 gcagccgcta aggctgctgc tggcggatct acaccctacc tgcagttcga tagaaaccag
     4321 tgggccgctc tgagagacag cgtgccaatg acactgagcg aggacgagat cgccagactg
     4381 aagggcatca atgaggacct gagcctggaa gaggtggccg agatttacct gcccctgagc
     4441 cggctgctga acttctacat cagcagcaac ctgcggagac aggccgtgct ggaacagttc
     4501 ctgggaacaa acggccagcg gatcccctac atcatctcta tcgccggatc tgtggccgtg
     4561 ggcaagagca caacagctag agttctgcag gccctgctga gcagatggcc cgagcataga
     4621 agagtggaac tgattaccac cgccggcttt ctgcacccca accaggtgct gaaagaacgg
     4681 ggcctgatga agaagaaggg cttccccgag agctacgaca tgcaccggct ggtcaagttt
     4741 gtgtccgacc tgaagtccgg cgtgcccaat gtgacagccc ctgtgtacag ccacctgatc
     4801 tacgacgtga tccccgacgg cgataagacc gtggtgcagc ccgatatcct gattctggaa
     4861 ggcgacaacg tgctgcagag cggcatggac tatcctcatg atcctcacca cgtgttcgtg
     4921 tccgacttcg tggacttcag catctacgtg gacgcccctg aggatctgct gcagacctgg
     4981 tacatcaacc ggtttctgaa gttcagagag ggcgcctaca cagaccccga cagctacttc
     5041 cacaactacg ccaagctgac caaagaggaa gccatcaaga ccgccatgac ctggtggaaa
     5101 gagatcaact ggctgaacct gaagcagaac atcctgccta ccagagagcg ggccagcctg
     5161 atcctgacca aatctgccaa tcacgccgtg gaagaagtgc ggctgagaaa aggcgctcca
     5221 ggattctcct caatttccgc ttaagcggcc gctcgagtct agagggcccg tttaaacccg
     5281 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     5341 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     5401 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     5461 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     5521 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     5581 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     5641 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     5701 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     5761 cgaccccaaa aaacttgatt agggtgatgg ttcacgtacc tagaagttcc tattccgaag
     5821 ttcctattct ctagaaagta taggaacttc cttggccaaa aagcctgaac tcaccgcgac
     5881 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc
     5941 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg
     6001 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc
     6061 ggccgcgctc ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta
     6121 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc
     6181 cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca
     6241 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga
     6301 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac
     6361 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc
     6421 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg
     6481 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt
     6541 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt
     6601 cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat
     6661 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc
     6721 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat
     6781 cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg
     6841 aaaccgacgc cccagcactc gtccgagggc aaaggaatag cacgtactac gagatttcga
     6901 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     6961 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     7021 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     7081 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     7141 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     7201 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     7261 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     7321 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     7381 cggtttgcgt attgg
//
