LOCUS       sequence-478603-        8591 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Contains miniCMV promoter expressing iRFP670 mRNA taged 24xMS2 
            motif.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8591)
  AUTHORS   Zhou WJ, Wu MY, Shao XJ, Tang LJ, Wang F, Jiang JH
  TITLE     Fluorogenic Interacting Protein Stabilization for Orthogonal RNA 
            Imaging.
  JOURNAL   Angew Chem Int Ed Engl. 2025 Apr 10:e202502350. doi: 
            10.1002/anie.202502350.
  PUBMED    40208703
REFERENCE   2  (bases 1 to 8591)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Angew Chem 
            Int Ed Engl. 2025 Apr 10:e202502350. doi: 10.1002/anie.202502350."
COMMENT     Sequence Label: miniCMV-iRFP670-24xMS2
FEATURES             Location/Qualifiers
     source          1..8591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2225..2263
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2260..2284
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2301..2310
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2307..3242
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     misc_feature    4981..5569
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5034..5054)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5452..5463)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5644..5877
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5996..6014)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6082..6186
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6187..6255,6256..7047)
                     /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(6405..6424)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7218..7806
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7707..7726
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7960..7977
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8217..8238
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8253..8258,8259..8276,8277..8283)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8291..8307
                     /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     8296..8318
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8315..8331
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8315..8331
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8381..8400
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtacg
     2221 cgttggtagg cgtgtacggt gggaggccta tataagcaga gctcgtttag tgaaccgtca
     2281 gatcgctcta gagcggccgc gccaccatgg cgcgtaaggt cgatctcacc tcctgcgatc
     2341 gcgagccgat ccacatcccc ggcagcattc agccgtgcgg ctgcctgcta gcctgcgacg
     2401 cgcaggcggt gcggatcacg cgcattacgg aaaatgccgg cgcgttcttt ggacgcgaaa
     2461 ctccgcgggt cggtgagcta ctcgccgatt acttcggcga gaccgaagcc catgcgctgc
     2521 gcaacgcact ggcgcagtcc tccgatccaa agcgaccggc gctgatcttc ggttggcgcg
     2581 acggcctgac cggccgcacc ttcgacatct cactgcatcg ccatgacggt acatcgatca
     2641 tcgagttcga gcctgcggcg gccgaacagg ccgacaatcc gctgcggctg acgcggcaga
     2701 tcatcgcgcg caccaaagaa ctgaagtcgc tcgaagagat ggccgcacgg gtgccgcgct
     2761 atctgcaggc gatgctcggc tatcaccgcg tgatgttgta ccgcttcgcg gacgacggct
     2821 ccgggatggt gatcggcgag gcgaagcgca gcgacctcga gagctttctc ggtcagcact
     2881 ttccggcgtc gctggtcccg cagcaggcgc ggctactgta cttgaagaac gcgatccgcg
     2941 tggtctcgga ttcgcgcggc atcagcagcc ggatcgtgcc cgagcacgac gcctccggcg
     3001 ccgcgctcga tctgtcgttc gcgcacctgc gcagcatctc gccctgccat ctcgaatttc
     3061 tgcggaacat gggcgtcagc gcctcgatgt cgctgtcgat catcattgac ggcacgctat
     3121 ggggattgat catctgtcat cattacgagc cgcgtgccgt gccgatggcg cagcgcgtcg
     3181 cggccgaaat gttcgccgac ttcttatcgc tgcacttcac cgccgcccac caccaacgct
     3241 gagctagcta ggcggactgt tactgagctg cgttttacac cctttctttg acaaaaccta
     3301 acttgcgcag aaaaaaaaaa aataagagac aacattggca tggctttgtt tttttaaatt
     3361 ttttttaaag tttttttttt tttttttttt ttttaagttt ttttgttttg ttttggcgct
     3421 tttgactcag gatttaaaaa ctggaacggt gaaggcgaca gcagttggtt ggagcaaaca
     3481 tcccccaaag ttctacaaat gtggctgagg actttgtaca ttgttttgtt tttttttttt
     3541 ttggttttgt ctttttttaa tagtcattcc aagtatccat gaaataagtg gttacaggaa
     3601 gtccctcacc ctcccaaaag ccacccccac tcctaagagg aggatggtcg cgtccatgcc
     3661 ctgagtccac cccggggaag gtgacaaccg gtaacctaca aacgggtgga ggatcacccc
     3721 acccgacact tcacaatcaa ggggtacaat acacaagggt ggaggaacac cccaccctcc
     3781 agacacatta cacagaaatc caatcaaaca gaagcaccat cagggcttct gctaccaaat
     3841 ttatctcaaa aaactacaac aaggaatcac catcagggat tccctgtgca atatacgtca
     3901 aacgagggcc acgacgggag gacgatcacg cctcccgaat atcggcatgt ctggctttcg
     3961 aattcagtgc gtggagcatc agcccacgca gccaatcaga gtcgaataca agtcgacttt
     4021 cgcgaagagc atcagccttc gcgccattct tacacaaacc acactctccc ctacaggaac
     4081 agcatcagcg ttcctgccca gtacccaact caagaaaatt tatgtcccca tgcagcatca
     4141 gcgcatgggc cccaagaata catccccaac aaaatcacat ccgagcacca acagggctcg
     4201 gagtgttgtt tcttgtccaa ctggacaaac cctccatgga ccatcaggcc atggactctc
     4261 accaacaaga caaaaactac tcttctcgaa gcagcatcag cgcttcgaaa cactcgagca
     4321 tacattgtgc ctatttcttg ggtggacgat cacgccaccc atgctctcac gaatttcaaa
     4381 acacggacaa ggacgagcac caccagggct cgtcgttcca cgtccaatac gattacttac
     4441 ctttcgggat cacgatcacg gatcccgcag ctacatcact tccactcagg acattcaagc
     4501 atgcacgatc acggcatgct ccacaagtct caaccacaga aactaccaaa tgggttcagc
     4561 accagcgaac ccactcctac ctcaaacctc ttcccacaaa actggcaagc aggatcaccg
     4621 cttgcccatt ccaacatacc aaatcaaaaa caattactgg tacagcatca gcgtaccagc
     4681 ccacatctct cactactatc aaaaaccaaa ccgttcagca acagcgaacg gtacacacgg
     4741 aaaaatcaac tggtttacaa atacgaaaga cgatcacgct ttcgtccagc gcaaactatt
     4801 acgaaaaaca tccgacggga agagcaacag ccttcccgcg gcggaaaacc tcacaaaaac
     4861 acgacaaacg gatgcacgaa cacggcatcc gccgacaacc cacaaactta caaccaggca
     4921 aacggtgcag gatcaccgca ccgtacatca aacacctcag atctcatatc gatagatcct
     4981 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     5041 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     5101 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     5161 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     5221 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     5281 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     5341 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     5401 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     5461 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     5521 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tgagatcctt
     5581 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     5641 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt
     5701 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     5761 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     5821 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     5881 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     5941 gtgagaggcc cgggttaatt aaggaaaggg ctagatcatt cttgaagacg aaagggcctc
     6001 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     6061 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     6121 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     6181 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     6241 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     6301 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     6361 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     6421 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     6481 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     6541 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     6601 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     6661 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6721 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     6781 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     6841 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6901 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6961 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     7021 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     7081 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     7141 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     7201 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     7261 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     7321 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     7381 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     7441 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     7501 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     7561 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     7621 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     7681 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7741 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     7801 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     7861 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     7921 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     7981 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     8041 agcaagctca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     8101 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     8161 tccccgtggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     8221 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     8281 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg
     8341 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     8401 aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa
     8461 cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc
     8521 tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca
     8581 aacttagtag t
//
