LOCUS       sequence-478602-        6475 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Contains iRFP670-fCse3 fluorogenic protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6475)
  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 6475)
  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: iRFP670-fCse3
FEATURES             Location/Qualifiers
     source          1..6475
                     /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"
     promoter        235..273
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     270..294
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             309..1241
                     /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"
     CDS             1281..1307
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             1317..1343
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     promoter        complement(2028..2046)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2029..2046)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2066..2083)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2072..2296
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2342..2770
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2429..2448)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2639..2660
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2779..2799)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2784..3113
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2964..3099
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3026..3045
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3180..3974
                     /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(3234..3253)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3844..3863
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4148..4269
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4185..4204)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4239..4258
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4318..4334)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4318..4334)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4331..4353)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4342..4358
                     /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(4366..4372,4373..4390,4391..4396))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4411..4432
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4549..4566)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4720..5308)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4800..4819)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5479..6270,6271..6339))
                     /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     6102..6121
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6340..6444)
                     /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 cgttggtagg
      241 cgtgtacggt gggaggccta tataagcaga gctcgtttag tgaaccgtca gatcgcgccg
      301 acatcaccat ggcgcgtaag gtcgatctca cctcctgcga tcgcgagccg atccacatcc
      361 ccggcagcat tcagccgtgc ggctgcctgc tagcctgcga cgcgcaggcg gtgcggatca
      421 cgcgcattac ggaaaatgcc ggcgcgttct ttggacgcga aactccgcgg gtcggtgagc
      481 tactcgccga ttacttcggc gagaccgaag cccatgcgct gcgcaacgca ctggcgcagt
      541 cctccgatcc aaagcgaccg gcgctgatct tcggttggcg cgacggcctg accggccgca
      601 ccttcgacat ctcactgcat cgccatgacg gtacatcgat catcgagttc gagcctgcgg
      661 cggccgaaca ggccgacaat ccgctgcggc tgacgcggca gatcatcgcg cgcaccaaag
      721 aactgaagtc gctcgaagag atggccgcac gggtgccgcg ctatctgcag gcgatgctcg
      781 gctatcaccg cgtgatgttg taccgcttcg cggacgacgg ctccgggatg gtgatcggcg
      841 aggcgaagcg cagcgacctc gagagctttc tcggtcagca ctttccggcg tcgctggtcc
      901 cgcagcaggc gcggctactg tacttgaaga acgcgatccg cgtggtctcg gattcgcgcg
      961 gcatcagcag ccggatcgtg cccgagcacg acgcctccgg cgccgcgctc gatctgtcgt
     1021 tcgcgcacct gcgcagcatc tcgccctgcc atctcgaatt tctgcggaac atgggcgtca
     1081 gcgcctcgat gtcgctgtcg atcatcattg acggcacgct atggggattg atcatctgtc
     1141 atcattacga gccgcgtgcc gtgccgatgg cgcagcgcgt cgcggccgaa atgttcgccg
     1201 acttcttatc gctgcacttc accgccgccc accaccaacg cctgtacaag ggaggcggag
     1261 gctccggagg cggaggctcc ctgccccccc tggagcgcct gaccctggga ggctccctgc
     1321 cccccctgga gcgcctgacc ctgggttccg gtggatccgg ttccggagtt gcgacagtca
     1381 ttaaaactaa acaggttgaa tttcaacttc aggttggtgt tccactctat tttcggcttc
     1441 gggcaaatcc gatcaaaact attctcgaca atcaaaagcg cctggacagt aaagggaata
     1501 ttaaacgctg tcgggttccg ttaataaaag aagcagaaca aatcgcgtgg ttgcaacgta
     1561 aattgggcaa tgcggcgcgc gttgaagatg tgcatcccat atcggaacgg ccacagtatt
     1621 tttctggtga tggtaaaagt ggaaagatcc aaacggtttg ctttgaaggt gtgctcacca
     1681 tcaacgacgc gccagcgtta atagatcttg tacagcaagg tattgggcca gctaaatcga
     1741 tgggatgtgg cttgctatct ttggctccac tgggctccgg aatgtatctc agtaaagtca
     1801 tcattgccag ggcctggagc agggatcttt accaacttgc ccagggatta tggcatttat
     1861 ttccaaacag accggatgct gctcgtgatt ttctttttca tgttgagaag cgaaacacac
     1921 cagaaggctg tcatgtttta ttgcagtcag cgcaaatgcc tgtttcaact gccggttccc
     1981 gtcgccgccg tcgtcgccgc ggttaagaat tctctagagg gccctattct atagtgtcac
     2041 ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2101 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2161 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2221 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     2281 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     2341 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2401 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2461 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2521 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2581 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2641 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2701 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2761 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     2821 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     2881 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2941 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3001 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3061 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3121 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     3181 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3241 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3301 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3361 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3421 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3481 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3541 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3601 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3661 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     3721 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     3781 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     3841 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     3901 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     3961 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4021 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4081 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4141 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4201 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4261 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4321 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4381 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4441 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4501 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4561 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4621 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4681 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4741 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4801 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4861 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4921 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4981 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5041 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5101 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5161 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5221 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5281 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5341 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5401 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5461 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5521 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5581 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5641 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5701 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5761 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5821 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5881 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5941 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6001 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6061 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6121 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6181 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6241 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6301 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6361 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6421 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
