LOCUS       sequence-387297-        7038 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 7038)
  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 7038)
  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_Halo-PanK
FEATURES             Location/Qualifiers
     source          1..7038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(165..184)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        356..735
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        736..939
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     890..910
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    941..959
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    962..980
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     990..1014
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1111..1120
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1117..2007
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     primer_bind     complement(3111..3128)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3117..3341
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3474..3493)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3625..3672
                     /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             3680..4702
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3680..4702
                     /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    4832..4953
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4869..4888)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4923..4942
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5002..5018)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5002..5018)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5015..5037)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5026..5042
                     /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(5050..5056,5057..5074,5075..5080))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5095..5116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5233..5250)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5404..5992)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5484..5503)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6163..6954,6955..7023))
                     /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     6786..6805
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7024..7038,1..90))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
       61 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
      121 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
      181 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
      241 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
      301 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
      361 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      421 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      481 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      541 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      601 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      661 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      721 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      781 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      841 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      901 ggtaggcgtg tacggtggga ggtctatata agcagagctc tccctatcag tgatagagat
      961 ctccctatca gtgatagaga tcgtcgacga gctcgtttag tgaaccgtca gatcgcctgg
     1021 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
     1081 actctagcgt ttaaacttaa gcttggtacc gccaccatgg cagagatcgg aaccggcttt
     1141 cctttcgacc ctcactacgt ggaagtgctg ggcgagagaa tgcactatgt ggacgtgggc
     1201 cccagagatg gaacccctgt gctgtttctg cacggcaacc ctaccagcag ctacgtgtgg
     1261 cggaacatca tccctcacgt ggcccctaca cacagatgta tcgcccctga cctgatcggc
     1321 atgggcaaga gcgataagcc cgacctgggc tacttcttcg acgaccacgt gcggttcatg
     1381 gacgccttta ttgaggccct cggcctggaa gaggtggtgc tggttattca cgattggggc
     1441 agcgccctgg gattccactg ggccaagaga aaccccgaga gagtgaaggg aatcgccttc
     1501 atggaattca tcaggcccat tcctacctgg gacgagtggc ctgagttcgc cagagagaca
     1561 ttccaggcct tcagaaccac cgacgtgggc agaaagctga tcatcgacca gaacgtgttc
     1621 atcgagggca ccctgcctat gggagtcgtt agacctctga ccgaggtgga aatggaccac
     1681 tacagagagc cctttctgaa ccccgtggac cgggaacctc tttggagatt ccccaacgag
     1741 ctgcccattg ccggcgagcc tgctaatatt gtggccctgg tggaagagta catggactgg
     1801 ctgcatcaga gccccgtgcc taagctgctg ttttggggaa cacccggcgt gctgattcct
     1861 cctgctgaag ctgccagact ggccaagagc ctgcctaatt gcaaggccgt ggatatcggc
     1921 cctggcctga atctgctgca agaggacaac cccgatctga tcggctccga gattgccaga
     1981 tggctgagca ccctggaaat cagcggagga tctctggctg aagccgccgc taaagaagcc
     2041 gctgcaaaag aggctgccgc caaagaggca gcagcaaaag aagcagccgc taaggctgct
     2101 gctggcggat ctacacccta cctgcagttc gatagaaacc agtgggccgc tctgagagac
     2161 agcgtgccaa tgacactgag cgaggacgag atcgccagac tgaagggcat caatgaggac
     2221 ctgagcctgg aagaggtggc cgagatttac ctgcccctga gccggctgct gaacttctac
     2281 atcagcagca acctgcggag acaggccgtg ctggaacagt tcctgggaac aaacggccag
     2341 cggatcccct acatcatctc tatcgccgga tctgtggccg tgggcaagag cacaacagct
     2401 agagttctgc aggccctgct gagcagatgg cccgagcata gaagagtgga actgattacc
     2461 accgccggct ttctgcaccc caaccaggtg ctgaaagaac ggggcctgat gaagaagaag
     2521 ggcttccccg agagctacga catgcaccgg ctggtcaagt ttgtgtccga cctgaagtcc
     2581 ggcgtgccca atgtgacagc ccctgtgtac agccacctga tctacgacgt gatccccgac
     2641 ggcgataaga ccgtggtgca gcccgatatc ctgattctgg aaggcgacaa cgtgctgcag
     2701 agcggcatgg actatcctca tgatcctcac cacgtgttcg tgtccgactt cgtggacttc
     2761 agcatctacg tggacgcccc tgaggatctg ctgcagacct ggtacatcaa ccggtttctg
     2821 aagttcagag agggcgccta cacagacccc gacagctact tccacaacta cgccaagctg
     2881 accaaagagg aagccatcaa gaccgccatg acctggtgga aagagatcaa ctggctgaac
     2941 ctgaagcaga acatcctgcc taccagagag cgggccagcc tgatcctgac caaatctgcc
     3001 aatcacgccg tggaagaagt gcggctgaga aaaggcgctc caggattctc ctcaatttcc
     3061 gcttaagcgg ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     3121 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3181 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3241 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3301 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3361 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3421 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3481 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3541 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3601 ttagggtgat ggttcacgta cctagaagtt cctattccga agttcctatt ctctagaaag
     3661 tataggaact tccttggcca aaaagcctga actcaccgcg acgtctgtcg agaagtttct
     3721 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg
     3781 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
     3841 tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc
     3901 ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc
     3961 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt
     4021 cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc
     4081 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
     4141 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc
     4201 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
     4261 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat
     4321 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
     4381 gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga
     4441 gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta
     4501 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc
     4561 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc
     4621 cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac
     4681 tcgtccgagg gcaaaggaat agcacgtact acgagatttc gattccaccg ccgccttcta
     4741 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4801 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4861 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4921 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4981 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5041 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5101 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5161 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5221 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5281 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5341 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5401 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5461 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5521 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5581 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5641 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5701 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5761 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5821 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5881 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5941 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6001 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6061 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6121 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6181 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6241 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6301 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6361 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6421 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6481 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6541 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6601 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6661 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6721 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6781 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6841 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6901 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6961 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7021 catactcttc ctttttca
//
