LOCUS       sequence-410730-        7642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express beta-ACTIN-3′UTR-8xMS2 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7642)
  AUTHORS   Hu Y, Xu J, Gao E, Fan X, Wei J, Ye B, Xu S, Ma W
  TITLE     Enhanced single RNA imaging reveals dynamic gene expression in live 
            animals.
  JOURNAL   Elife. 2023 Mar 3;12:e82178. doi: 10.7554/eLife.82178.
  PUBMED    36867026
REFERENCE   2  (bases 1 to 7642)
  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: "Elife."; 
            date: "2023-03-3"; volume: "12:e82178. doi"; pages: " 
            10.7554/eLife.82178"
COMMENT     Sequence Label: pcDNA-puro-TagBFP-beta-ACTIN-3′UTR-8xMS2
FEATURES             Location/Qualifiers
     source          1..7642
                     /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"
     CDS             905..1603
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_RNA        3147..3165
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3186..3204
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3231..3249
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3270..3288
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3315..3333
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3354..3372
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3399..3417
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3438..3456
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     primer_bind     complement(3482..3499)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3488..3712
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3758..4186
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3845..3864)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4055..4076
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4195..4215)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4200..4529
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4380..4515
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4442..4461
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4542..5141
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4542..4561)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5038..5058
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5315..5436
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5352..5371)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5406..5425
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5485..5501)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5485..5501)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5498..5520)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5509..5525
                     /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(5533..5539,5540..5557,5558..5563))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5578..5599
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5716..5733)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5887..6475)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5967..5986)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6646..7437,7438..7506))
                     /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     7269..7288
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7507..7611)
                     /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 aaccatgagc gagctgatta aggagaacat gcacatgaag ctctacatgg agggcaccgt
      961 ggacaaccat cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca
     1021 gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc
     1081 tactagcttc ctctacggca gcaagacctt catcaaccac acccagggca tccccgactt
     1141 cttcaagcag tccttccctg agggcttcac atgggagaga gtcaccacat acgaagacgg
     1201 gggcgtgctg accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt
     1261 caagatcaga ggggtgaact tcacatccaa cggccctgtg atgcagaaga aaacactcgg
     1321 ctgggaggcc ttcaccgaga cgctgtaccc cgctgacggc ggcctggaag gcagaaacga
     1381 catggccctg aagctcgtgg gcgggagcca tctgatcgca aacatcaaga ccacatatag
     1441 atccaagaaa cccgctaaga acctcaagat gcctggcgtc tactatgtgg actacagact
     1501 ggaaagaatc aaggaggcca acaacgagac ctacgtcgag cagcacgagg tggcagtggc
     1561 cagatactgc gacctcccta gcaaactggg gcacaagctg aatctgtaca agaagcttgg
     1621 taccgatgat gatatcgccg cgctcgtcgt cgacaacggc tccggcatgt gcaaggccgg
     1681 cttcgcgggc gacgatgccc cccgggccgt cttcccctcc atcgtggggc gccccaggca
     1741 ccagggcgtg atggtgggca tgggtcagaa ggattcctat gtgggcgacg aggcccagag
     1801 caagagaggc atcctcaccc tgaagtaccc catcgagcac ggcatcgtca ccaactggga
     1861 cgacatggag aaaatctggc accacacctt ctacaatgag ctgcgtgtgg ctcccgagga
     1921 gcaccccgtg ctgctgaccg aggcccccct gaaccccaag gccaaccgcg agaagatgac
     1981 ccagatcatg tttgagacct tcaacacccc agccatgtac gttgctatcc aggctgtgct
     2041 atccctgtac gcctctggcc gtaccactgg catcgtgatg gactccggtg acggggtcac
     2101 ccacactgtg cccatctacg aggggtatgc cctcccccat gccatcctgc gtctggacct
     2161 ggctggccgg gacctgactg actacctcat gaagatcctc accgagcgcg gctacagctt
     2221 caccaccacg gccgagcggg aaatcgtgcg tgacattaag gagaagctgt gctacgtcgc
     2281 cctggacttc gagcaagaga tggccacggc tgcttccagc tcctccctgg agaagagcta
     2341 cgagctgcct gacggccagg tcatcaccat tggcaatgag cggttccgct gccctgaggc
     2401 actcttccag ccttccttcc tgggcatgga gtcctgtggc atccacgaaa ctaccttcaa
     2461 ctccatcatg aagtgtgacg tggacatccg caaagacctg tacgccaaca cagtgctgtc
     2521 tggcggcacc accatgtacc ctggcattgc cgacaggatg cagaaggaga tcactgccct
     2581 ggcacccagc acaatgaaga tcaagatcat tgctcctcct gagcgcaagt actccgtgtg
     2641 gatcggcggc tccatcctgg cctcgctgtc caccttccag cagatgtgga tcagcaagca
     2701 ggagtatgac gagtccggcc cctccatcgt ccaccgcaaa tgcttctagg cggactatga
     2761 cttagttgcg ttacaccctt tcttgacaaa acctaacttg cgcagaaaac aagatgagat
     2821 tggcatggct ttatttgttt tttttgtttt gttttggttt tttttttttt tggcttgact
     2881 caggatttaa aaactggaac ggtgaaggtg acagcagtcg gttggagcga gcatccccca
     2941 aagttcacaa tgtggccgag gactttgatt gcacattgtt gtttttttaa tagtcattcc
     3001 aaatatgaga tgcattgtta caggaagtcc cttgccatcc taaaagccac cccacttctc
     3061 tctaaggaga atggcccagt cctctcccaa gtccacacag gggaggtgat agcattgctt
     3121 ggatccctgc aggtcgactc tagaaaacat gaggatcacc catgtctgca ggtcgactct
     3181 agaaaacatg aggatcaccc atgtgaattc ctgcaggtcg actctagaaa acatgaggat
     3241 cacccatgtc tgcaggtcga ctctagaaaa catgaggatc acccatgtga tatcctgcag
     3301 gtcgactcta gaaaacatga ggatcaccca tgtctgcagg tcgactctag aaaacatgag
     3361 gatcacccat gtctcgagct gcaggtcgac tctagaaaac atgaggatca cccatgtctg
     3421 caggtcgact ctagaaaaca tgaggatcac ccatgtgggc ccgtttaaac ccgctgatca
     3481 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3541 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3601 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3661 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     3721 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     3781 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3841 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3901 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3961 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4021 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4081 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4141 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4201 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4261 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4321 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4381 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4441 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4501 gcttttttgg aggcctaggc ttttgcaaaa agctcccaac catgaccgag tacaagccca
     4561 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     4621 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     4681 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     4741 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     4801 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     4861 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     4921 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     4981 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     5041 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     5101 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg agcgggactc tggggttcga
     5161 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5221 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5281 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5341 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5401 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5461 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5521 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5581 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5641 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5701 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5761 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5821 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5881 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5941 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6001 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6061 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6121 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6181 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6241 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6301 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6361 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6421 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6481 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6541 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6601 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6661 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6721 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6781 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6841 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6901 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6961 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7021 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7081 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7141 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7201 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7261 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7321 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7381 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7441 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7501 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7561 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7621 ccgaaaagtg ccacctgacg tc
//
