LOCUS       sequence-366482-        6135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express histone H2B tagged with miniTurbo under the control of CAG 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6135)
  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 6135)
  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-H2B-mTurbo
FEATURES             Location/Qualifiers
     source          1..6135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..663
                     /label=chicken beta-actin promoter
     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"
     misc_feature    3001..3057
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(3157..3176)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3222..3277
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3276..3295)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3638..3654)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3638..3654)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3651..3673)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3662..3678
                     /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(3686..3692,3693..3710,3711..3716))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3731..3752
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3811..4006
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3857..3992
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4012..4146
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4049..4068)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4103..4122
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4214..4231)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4385..4973)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4465..4484)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5144..5935,5936..6004))
                     /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     5767..5786
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6005..6109)
                     /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 gggsggggsg gggsgrgggg
      541 sggggsgggg sgrggcggag 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 atgccagagc cagcgaagtc tgctcccgcg cccaagaagg gctcgaagaa agccgtgact
     1801 aaggcgcaga agaaggacgg caagaagcgc aagcgcagcc gcaaggagag ctactccgta
     1861 tacgtgtaca aggtgctgaa gcaggtccac cccgacaccg gcatctcctc taaggccatg
     1921 ggaatcatga actccttcgt caacgacatc ttcgaacgca tcgcgggtga ggcttcccgc
     1981 ctggcgcatt acaacaagcg ctcgaccatc acctccaggg agatccagac ggccgtgcgc
     2041 ctgctgctgc ccggggagtt ggccaagcac gccgtgtccg agggcaccaa ggccatcacc
     2101 aagtacacca gcgctaagat ggcctctatg actggtggtc aacaaatggg ccggggaagc
     2161 gaatttatga ttcctcttct gaacgcaaag caaatactgg gtcagctcga cgggggatca
     2221 gtcgctgttt tgcctgttgt tgactctact aatcaatatc tcttggatcg aatcggtgaa
     2281 ctcaaatctg gcgatgcctg tatcgccgaa taccaacagg caggtagagg ttctagaggg
     2341 agaaaatggt tctccccgtt tggtgccaat ctctatttga gtatgttttg gcggcttaag
     2401 cgaggaccgg cggctatcgg tctcggacct gttattggca tcgtcatggc cgaggcgctt
     2461 aggaaacttg gagccgataa agtcagggtc aagtggccaa atgatttgta tttgcaagac
     2521 agaaagctcg caggcatctt ggttgagctt gctgggataa cgggcgacgc agcacaaata
     2581 gtcattggtg ccggaatcaa tgtcgctatg cgccgggttg aagaaagtgt cgtgaaccaa
     2641 ggttggataa ctcttcaaga agcgggtatt aacctcgatc gcaatactct cgccgctatg
     2701 ctcatcaggg aactgcgagc ggccctggag ttgttcgagc aagaaggtct cgctccctat
     2761 ttgtcacgct gggaaaagtt ggataatttc attaaccgac cagtcaaact tattattggc
     2821 gataaagaaa tatttgggat ctcacgcggt atcgataagc aaggcgcatt gttgcttgaa
     2881 caggatggag tgattaaacc atggatggga ggtgagatca gccttagaag tgcggaaaag
     2941 aaatccggac tcaccggtgg aggcggttca ggcggaggtg gctctggcgg tggcggatcg
     3001 agatctcgag ctcaagcttc gaattctgca gtcgacggta ccgcgggccc gggatccacc
     3061 ggatctagat aactgatcaa ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg
     3121 tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta
     3181 tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt
     3241 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     3301 aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat
     3361 gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc
     3421 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
     3481 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
     3541 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag
     3601 atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     3661 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     3721 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     3781 agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc
     3841 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3901 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3961 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt
     4021 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4081 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4141 tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4201 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4261 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4321 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4381 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4441 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4501 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4561 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4621 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4681 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4741 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4801 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4861 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4921 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4981 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5041 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5101 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5161 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5221 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5281 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5341 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5401 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5461 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5521 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5581 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5641 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5701 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5761 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5821 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5881 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5941 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6001 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6061 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6121 gaaaagtgcc acctg
//
