LOCUS       sequence-481928-        5906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the luminescent protein nLuc fused to the encapsulation 
            signal for QtEnc and fused to a degron signal for proteasomal 
            degradation when not encapsulated..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5906)
  AUTHORS   Berezin O, Piovesan A, Graf R, Samara E, Sigmund F, Westmeyer GG
  TITLE     Multiplexed genetic tags for electron and fluorescence microscopy.
  JOURNAL   Nat Protoc. 2025 Nov 19. doi: 10.1038/s41596-025-01260-7.
  PUBMED    41261217
REFERENCE   2  (bases 1 to 5906)
  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 Protoc.
            2025 Nov 19. doi: 10.1038/s41596-025-01260-7."
COMMENT     Sequence Label: DD-N-nLuc-QtSig
FEATURES             Location/Qualifiers
     source          1..5906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      958..967
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(964..966,967..1287)
                     /codon_start=1
                     /product="destabilization domain that can be stabilized by 
                     Shield1 in the ProteoTuner(TM) system"
                     /label=DD
                     /note="L106P mutant of FKBP12"
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKPE"
     primer_bind     complement(1914..1931)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1920..2144
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2190..2618
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2277..2296)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2487..2508
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2627..2647)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2632..2961
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2812..2947
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2874..2893
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3009..3056
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3075..3449
                     /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"
     polyA_signal    3579..3700
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3616..3635)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3670..3689
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3749..3765)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3749..3765)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3762..3784)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3773..3789
                     /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(3797..3803,3804..3821,3822..3827))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3842..3863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3980..3997)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4151..4739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4231..4250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4910..5701,5702..5770))
                     /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     5533..5552
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5771..5875)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatgggag tgcaggtgga aaccatctcc ccaggagacg ggcgcacctt ccccaagcgc
     1021 ggccagacct gtgtggtgca ctacaccggg atgcttgaag atggaaagaa agtcgattcc
     1081 tcccgggaca gaaacaagcc ctttaagttt atgctaggca agcaggaggt gatccgaggc
     1141 tgggaagaag gggttgccca gatgagtgtg ggtcagagag ccaaactgac tatatctcca
     1201 gattatgcct atggtgccac tgggcaccca ggcatcatcc caccacatgc cactctcgtc
     1261 ttcgatgtgg agcttctaaa accggaagga tctggggtgt tcaccctgga agatttcgtg
     1321 ggcgactggc ggcagaccgc tggctacaat ctggaccagg tgctggaaca gggcggcgtg
     1381 tcctctctgt ttcagaacct gggcgtgtcc gtgaccccca tccagagaat cgtgctgagc
     1441 ggcgagaacg gcctgaagat cgacatccac gtgatcatcc cttacgaggg cctgtccggc
     1501 gaccagatgg gccagatcga gaagatcttt aaggtggtgt accccgtgga cgaccaccac
     1561 ttcaaagtga tcctgcacta cggcaccctc gtgatcgacg gcgtgacccc taacatgatc
     1621 gactacttcg gcagacccta cgagggaatc gccgtgttcg acggcaagaa aatcaccgtg
     1681 accggcaccc tgtggaacgg caacaagatc atcgacgaga gactgatcaa ccccgacggc
     1741 agcctgctgt tcagagtgac catcaatggc gtgacaggct ggcggctgtg cgagagaatc
     1801 ctggccggcg gcggaggatc tggcggaggc ggatctaagg gcttcaccgt gggcagcctg
     1861 atccaatgag cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac
     1921 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     1981 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2041 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2101 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2161 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     2221 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2281 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2341 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2401 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2461 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2521 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2581 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     2641 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2701 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2761 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     2821 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     2881 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     2941 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3001 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     3061 tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg
     3121 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg
     3181 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
     3241 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg
     3301 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg
     3361 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg
     3421 tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg
     3481 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     3541 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     3601 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3661 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     3721 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     3781 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3841 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3901 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3961 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4021 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4081 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4141 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4201 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4261 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4321 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4381 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4441 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4501 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4561 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4621 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4681 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4741 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4801 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4861 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4921 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4981 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5041 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5101 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5161 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5221 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5281 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5341 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5401 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5461 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5521 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5581 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5641 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5701 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5761 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5821 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5881 ttccccgaaa agtgccacct gacgtc
//
