LOCUS       sequence-182613-        7844 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7844)
  AUTHORS   Wu B, Chao JA, Singer RH
  TITLE     Fluorescence fluctuation spectroscopy enables quantitative imaging 
            of single mRNAs in living cells.
  JOURNAL   Biophys J. 2012 Jun 20;102(12):2936-44. Epub 2012 Jun 19.
  PUBMED    22735544
REFERENCE   2  (bases 1 to 7844)
  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: "Biophys 
            J."; date: "2012-06-20"; volume: "102(12)"; pages: "2936-44. Epub 
            2012 Jun 19"
COMMENT     Sequence Label: phage-cmv-cfp-24xms2
FEATURES             Location/Qualifiers
     source          1..7844
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        25..43
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        64..82
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        134..152
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        173..191
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        243..261
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        282..300
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        352..370
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        391..409
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        461..479
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        500..518
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        570..588
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        609..627
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_feature    676..1264
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(729..749)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1147..1158)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1339..1572
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(1691..1709)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1777..1881
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1882..1950,1951..2742)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2100..2119)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2913..3501
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3402..3421
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3655..3672
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3912..3933
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3948..3953,3954..3971,3972..3978)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3986..4002
                     /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)."
     primer_bind     3991..4013
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4010..4026
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4010..4026
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4076..4095
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             4287..4920
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    4967..5092
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5589..5822
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6007..6051
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    6344..6461
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        6513..6816
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6817..7020
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6971..6991
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7017..7041
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      7107..7116
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7113..7115,7116..7118,7119..7832)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7158..7179)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7419..7438)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7766..7787
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ctaaggtacc taattgccta gaaaacatga ggatcaccca tgtctgcagg tcgactctag
       61 aaaacatgag gatcacccat gtctgcagta ttcccgggtt cattagatcc taaggtacct
      121 aattgcctag aaaacatgag gatcacccat gtctgcaggt cgactctaga aaacatgagg
      181 atcacccatg tctgcagtat tcccgggttc attagatcct aaggtaccta attgcctaga
      241 aaacatgagg atcacccatg tctgcaggtc gactctagaa aacatgagga tcacccatgt
      301 ctgcagtatt cccgggttca ttagatccta aggtacctaa ttgcctagaa aacatgagga
      361 tcacccatgt ctgcaggtcg actctagaaa acatgaggat cacccatgtc tgcagtattc
      421 ccgggttcat tagatcctaa ggtacctaat tgcctagaaa acatgaggat cacccatgtc
      481 tgcaggtcga ctctagaaaa catgaggatc acccatgtct gcagtattcc cgggttcatt
      541 agatcctaag gtacctaatt gcctagaaaa catgaggatc acccatgtct gcaggtcgac
      601 tccagaaaac atgaggatca cccatgtctg cagtattccc gggttcatta gatctgcgcg
      661 cgatcgatag atcctaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
      721 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
      781 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
      841 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
      901 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
      961 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     1021 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     1081 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     1141 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     1201 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     1261 ccgcctgaga tcctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     1321 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt
     1381 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     1441 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     1501 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     1561 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     1621 aatgaatatc agagagtgag aggcccgggt taattaagga aagggctaga tcattcttga
     1681 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     1741 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1801 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     1861 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     1921 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     1981 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     2041 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     2101 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     2161 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     2221 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     2281 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     2341 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     2401 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     2461 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     2521 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     2581 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     2641 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2701 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     2761 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     2821 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     2881 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2941 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3001 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     3061 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3121 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3181 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3241 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3301 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3361 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3421 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3481 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3541 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3601 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3661 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     3721 ccgattcatt aatgcagcaa gctcatggct gactaatttt ttttatttat gcagaggccg
     3781 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3841 gcttttgcaa aaagctcccc gtggcacgac aggtttcccg actggaaagc gggcagtgag
     3901 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     3961 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     4021 tatgacatga ttacgaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4081 ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac
     4141 caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt
     4201 catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt
     4261 taaatatgta ctacaaactt agtagttgga agggctaatt cactcccaaa gaagacaaga
     4321 tatccttgat ctgtggatct accacacaca aggctacttc cctgattagc agaactacac
     4381 accagggcca ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt
     4441 tgagccagat aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt
     4501 gagcctgcat gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg
     4561 cctagcattt catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata
     4621 tcgagcttgc tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg
     4681 gactggggag tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg
     4741 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     4801 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     4861 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     4921 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag
     4981 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
     5041 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
     5101 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
     5161 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
     5221 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
     5281 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
     5341 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
     5401 aaacaaaagt aagaccaccg cacagcaagc ggccggccgc tgatcttcag acctggagga
     5461 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     5521 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     5581 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     5641 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     5701 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     5761 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     5821 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     5881 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     5941 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     6001 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     6061 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     6121 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     6181 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     6241 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatct
     6301 cgacggtatc gccgaattca caaatggcag tattcatcca caattttaaa agaaaagggg
     6361 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     6421 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     6481 gcagagatcc agtttggact agtggagttc cgcgttacat aacttacggt aaatggcccg
     6541 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     6601 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     6661 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     6721 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     6781 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     6841 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     6901 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     6961 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     7021 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
     7081 agacaccggc ggccgcacca ccggtcgcca ccatggtgag caagggcgag gagctgttca
     7141 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     7201 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     7261 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tggggcgtgc
     7321 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     7381 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     7441 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     7501 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac atcagccaca
     7561 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
     7621 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     7681 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     7741 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     7801 tcactctcgg catggacgag ctgtacaagt aaagcggccg gatc
//
