LOCUS       sequence-366479-        5757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express miniTurbo under the control of CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5757)
  AUTHORS   Sanada T, Yamaguchi J, Furuta Y, Kakuta S, Tanida I, Uchiyama Y
  TITLE     In-resin CLEM of Epon-embedded cells using proximity labeling.
  JOURNAL   Sci Rep. 2022 Jul 1;12(1):11130. doi: 10.1038/s41598-022-15438-6.
  PUBMED    35778550
REFERENCE   2  (bases 1 to 5757)
  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: "Sci Rep."; 
            date: "2022-07-1"; volume: "12(1):11130. doi"; pages: " 
            10.1038/s41598-022-15438-6"
COMMENT     Sequence Label: pCAG-mTurbo-G
FEATURES             Location/Qualifiers
     source          1..5757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          664..1680
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1604..1626
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1688..1707
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1735..1744
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2623..2679
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(2779..2798)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2844..2899
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2898..2917)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3260..3276)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3260..3276)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3273..3295)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3284..3300
                     /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(3308..3314,3315..3332,3333..3338))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3353..3374
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3433..3628
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3479..3614
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3634..3768
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3671..3690)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3725..3744
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3836..3853)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4007..4595)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4087..4106)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4766..5557,5558..5626))
                     /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     5389..5408
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5627..5731)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gcgccrggsg gggggggggg gggsgggggg
      541 sggggsgggg sgrggsggrg aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
      601 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
      661 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
      721 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
      781 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
      841 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag
      961 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc
     1021 gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt
     1081 gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg
     1141 cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg
     1201 cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg
     1261 gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc
     1321 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
     1381 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
     1441 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
     1501 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt
     1561 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
     1621 gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag
     1681 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaagc tagcgccacc
     1741 atggcctcta tgactggtgg tcaacaaatg ggccggggaa gcgaatttat gattcctctt
     1801 ctgaacgcaa agcaaatact gggtcagctc gacgggggat cagtcgctgt tttgcctgtt
     1861 gttgactcta ctaatcaata tctcttggat cgaatcggtg aactcaaatc tggcgatgcc
     1921 tgtatcgccg aataccaaca ggcaggtaga ggttctagag ggagaaaatg gttctccccg
     1981 tttggtgcca atctctattt gagtatgttt tggcggctta agcgaggacc ggcggctatc
     2041 ggtctcggac ctgttattgg catcgtcatg gccgaggcgc ttaggaaact tggagccgat
     2101 aaagtcaggg tcaagtggcc aaatgatttg tatttgcaag acagaaagct cgcaggcatc
     2161 ttggttgagc ttgctgggat aacgggcgac gcagcacaaa tagtcattgg tgccggaatc
     2221 aatgtcgcta tgcgccgggt tgaagaaagt gtcgtgaacc aaggttggat aactcttcaa
     2281 gaagcgggta ttaacctcga tcgcaatact ctcgccgcta tgctcatcag ggaactgcga
     2341 gcggccctgg agttgttcga gcaagaaggt ctcgctccct atttgtcacg ctgggaaaag
     2401 ttggataatt tcattaaccg accagtcaaa cttattattg gcgataaaga aatatttggg
     2461 atctcacgcg gtatcgataa gcaaggcgca ttgttgcttg aacaggatgg agtgattaaa
     2521 ccatggatgg gaggtgagat cagccttaga agtgcggaaa agaaatccgg actcaccggt
     2581 ggaggcggtt caggcggagg tggctctggc ggtggcggat cgagatctcg agctcaagct
     2641 tcgaattctg cagtcgacgg taccgcgggc ccgggatcca ccggatctag ataactgatc
     2701 aactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     2761 tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc
     2821 ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     2881 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca
     2941 gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa
     3001 aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt
     3061 ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt
     3121 ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc
     3181 ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc
     3241 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     3301 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     3361 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3421 gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3481 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3541 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3601 cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt
     3661 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3721 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat
     3781 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     3841 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3901 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3961 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4021 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4081 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4141 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4201 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4261 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4321 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4381 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4441 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4501 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4561 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4621 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4681 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4741 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4801 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4861 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4921 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4981 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5041 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5101 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5161 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5221 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5281 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5341 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5401 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5461 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5521 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5581 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5641 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5701 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctg
//
