LOCUS       sequence-331476-        7171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  optimized miniCMV (OminiCMV).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7171)
  AUTHORS   Gao N, Hu J, He B, Ji Z, Hu X, Huang J, Wei Y, Peng J, Wei Y, Zhou 
            Y, Shen X, Li H, Feng X, Xiao Q, Shi L, Sun Y, Zhou C, Zhou H, Yang 
            H
  TITLE     Endogenous promoter-driven sgRNA for monitoring the expression of 
            low-abundance transcripts and lncRNAs.
  JOURNAL   Nat Cell Biol. 2021 Jan;23(1):99-108. doi: 
            10.1038/s41556-020-00610-9. Epub 2021 Jan 4.
  PUBMED    33398178
REFERENCE   2  (bases 1 to 7171)
  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"; doi: 
            "10.1038/s41556-020-00610-9"; journalName: "Nat Cell Biol"; date: 
            "2021-01"; volume: "23"; issue: "1"; pages: "99-108"
COMMENT     Sequence Label: PB-3XpolyA-3XPC-miniCMV-mCherry-WPRE-insulator
FEATURES             Location/Qualifiers
     source          1..7171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    8..129
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    141..262
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   557..591
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(620..636)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(620..636)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(633..655)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    644..660
                     /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(668..674,675..692,693..698))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    713..734
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(851..868)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1022..1610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1102..1121)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1781..2572,2573..2641))
                     /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     2404..2423
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(2659..3453)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2770..2789)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3380..3399
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      complement(3797..4225)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3907..3928)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4119..4138
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4351..4373
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4365..4382
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4366..4382
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4389..4408
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4389..4407
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   4493..4555
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    5202..5426
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5415..5432
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    5468..6056
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             5574..5585
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     5983..6003
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6062..6772)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(6340..6359)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6609..6627
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      6769..6778
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6792..6816)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(6813..6851)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     polyA_signal    7033..7167
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
       61 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
      121 aaacaagttt ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
      181 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
      241 tataagctgc aataaacaag ttccgcaccg gcgttaatta atgatatcta taacaagaaa
      301 atatatatat aataagttat cacgtaagta gaacatgaaa taacaatata attatcgtat
      361 gagttaaatc ttaaaagtca cgtaaaagat aatcatgcgt cattttgact cacgcggtcg
      421 ttatagttca aaatcagtga cacttaccgc attgacaagc acgcctcacg ggagctccaa
      481 gcggcgactg agatgtccta aatgcacagc gacggattcg cgctatttag aaagagagag
      541 caatatttca agaatgcatg cgtcaatttt acgcagacta tctttctagg gttaaggtac
      601 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
      661 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
      721 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
      781 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
      841 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
      901 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      961 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1021 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1081 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1141 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1201 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1261 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1321 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1381 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1441 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     1501 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1561 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1621 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1681 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1741 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1801 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1861 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1921 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1981 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     2041 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2101 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     2161 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     2221 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     2281 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     2341 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     2401 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     2461 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     2521 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     2581 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2641 tactcttcct ttttcaattc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg
     2701 cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag
     2761 ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag
     2821 ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca
     2881 ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc
     2941 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
     3001 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
     3061 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
     3121 ctcggcagga gcaaggtggg atgacaggag atcctgcccc ggcacttcgc ccaatagcag
     3181 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
     3241 ggccagccac gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc
     3301 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
     3361 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
     3421 agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg
     3481 atcagatctt gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac
     3541 tttgcagggc ttcccaacct taccagaggg cgccccagct ggcaattccg gttcgcttgc
     3601 tgtccataaa accgcccagt ctagctatcg ccatgtaagc ccactgcaag ctacctgctt
     3661 tctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatccgggg
     3721 tcagcaccgt ttctgcggac tggctttcta cgtgttccgc ttcctttagc agcccttgcg
     3781 ccctgaattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3841 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3901 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3961 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     4021 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4081 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4141 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4201 cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac tgttgggaag
     4261 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     4321 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     4381 gtgaattgta atacgactca ctatagggcg aattgggccc tctagatgca tgctcgagcg
     4441 gccgccagtg tgatggatat ctgcagaatt cgcccttatt ggccggcctt aaccctagaa
     4501 agataatcat attgtgacgt acgttaaaga taatcatgcg taaaattgac gcatgtgttt
     4561 tatcggtctg tatatcgagg tttatttatt aatttgaata gatattaagt tttattatat
     4621 ttacacttac atactaataa taaattcaac aaacaattta tttatgttta tttatttatt
     4681 aaaaaaaaac aaaaactcaa aatttcttct ataaagtaac aaaactttta ggcgcgtttt
     4741 ccccgtatcc ccccaggtgt ctgcaggctc aaagagcagc gagaagcgtt cagaggaaag
     4801 cgatcccgtg ccaccttccc cgtgcccggg ctgtccccgc acgctgccgg ctcggggatg
     4861 cggggggagc gccggaccgg agcggagccc cgggcggctc gctgctgccc cctagcgggg
     4921 gagggacgta attacatccc tgggggcttt gggggggggc tgtccctctt ttccccgtat
     4981 ccccccaggt gtctgcaggc tcaaagagca gcgagaagcg ttcagaggaa agcgatcccg
     5041 tgccaccttc cccgtgcccg ggctgtcccc gcacgctgcc ggctcgggga tgcgggggga
     5101 gcgccggacc ggagcggagc cccgggcggc tcgctgctgc cccctagcgg gggagggacg
     5161 taattacatc cctgggggct ttgggggggg gctgtccctc gccatagagc ccaccgcatc
     5221 cccagcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc
     5281 ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga
     5341 aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac
     5401 agatggctgg caactagaag gcacagtcga ggctgatcag cgggtttaaa cgggccccta
     5461 accggtggcg gggaggcggc ccaaagggag atccgactcg tctgagggcg aaggcggaga
     5521 cgcggaagag gccgcagagc cggcagcagg ccgcgggaag gaaggtccgc tggattgagg
     5581 gccgaaggga cgtagcagaa ggacgtcccg cgcagaatcc aggtggcaac acaggcgagc
     5641 agccaaggaa aggacgatga tttccccgac aacaccacgg aattgtcagt gcccaacagc
     5701 cgagcccctg tccagcagcg ggcaaggcag gcggcgatga gttccgccgt ggcaataggg
     5761 agggggaaag cgaaagtccc ggaaaggagc tgacaggtgg tggcaatgcc ccaaccagtg
     5821 ggggttgcgt cagcaaacac agtgcacacc acgccacgtt gcctgacaac gggccacaac
     5881 tcctcataaa gagacagcaa ccaggattta tacaaggagg agaaaatgaa agccatacgg
     5941 gaagcaatag catgatacaa aggcattaaa gcagcgtatc cacatagcgt aaaaggagca
     6001 acatagttaa gaataccagt caatctttca caaattttgt aatccagagg ttgattccgg
     6061 tttacttgta cagctcgtcc atgccgccgg tggagtggcg gccctcggcg cgttcgtact
     6121 gttccacgat ggtgtagtcc tcgttgtggg aggtgatgtc caacttgatg ttgacgttgt
     6181 aggcgccggg cagctgcacg ggcttcttgg ccttgtaggt ggtcttgacc tcagcgtcgt
     6241 agtggccgcc gtccttcagc ttcagcctct gcttgatctc gcccttcagg gcgccgtcct
     6301 cggggtacat ccgctcggag gaggcctccc agcccatggt cttcttctgc attacggggc
     6361 cgtcggaggg gaagttggtg ccgcgcagct tcaccttgta gatgaactcg ccgtcctgca
     6421 gggaggagtc ctgggtcacg gtcaccacgc cgccgtcctc gaagttcatc acgcgctccc
     6481 acttgaagcc ctcggggaag gacagcttca agtagtcggg gatgtcggcg gggtgcttca
     6541 cgtaggcctt ggagccgtac atgaactgag gggacaggat gtcccaggcg aagggcaggg
     6601 ggccaccctt ggtcaccttc agcttggcgg tctgggtgcc ctcgtagggg cggccctcgc
     6661 cctcgccctc gatctcgaac tcgtggccgt tcacggagcc ctccatgtgc accttgaagc
     6721 gcatgaactc cttgatgatg gccatgttat cctcctcgcc cttgctcacc atggtggcac
     6781 gcgttccagg cgatctgacg gttcactaaa cgagctctgc ttatataggc ctcccaccgt
     6841 acacgcctac cgctagctac gtccccactg tggggtggag gggacgtaaa ccctgcacgc
     6901 tcagttctcg ttggatacgt ccccactgtg gggtggaggg gacgtaaact cttctacgat
     6961 atagtccatt tgttacgtcc ccactgtggg gtggagggga cgtaaacgga tcccgattaa
     7021 tagtcgtcct ccgatccaga catgataaga tacattgatg agtttggaca aaccacaact
     7081 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     7141 accattataa gctgcaataa acaagtttca g
//
