LOCUS       sequence-478592-        6250 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Contains EGFP-fCse3 fluorogenic protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6250)
  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 6250)
  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: EGFP-fCse3
FEATURES             Location/Qualifiers
     source          1..6250
                     /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             join(309..311,312..314,315..1025)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(309..311,312..314,315..1025)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(354..375)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(615..634)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     962..983
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1056..1082
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             1092..1118
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     promoter        complement(1803..1821)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1804..1821)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1841..1858)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1847..2071
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2117..2545
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2204..2223)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2414..2435
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2554..2574)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2559..2888
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2739..2874
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2801..2820
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2955..3749
                     /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(3009..3028)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3619..3638
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3923..4044
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3960..3979)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4014..4033
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4093..4109)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4093..4109)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4106..4128)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4117..4133
                     /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(4141..4147,4148..4165,4166..4171))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4186..4207
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4324..4341)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4495..5083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4575..4594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5254..6045,6046..6114))
                     /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     5877..5896
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6115..6219)
                     /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 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      361 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      421 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      481 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
      541 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      601 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      661 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      721 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
      781 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
      841 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      901 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
      961 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1021 acaagggagg cggaggctcc ggaggcggag gctccctgcc ccccctggag cgcctgaccc
     1081 tgggaggctc cctgcccccc ctggagcgcc tgaccctggg ttccggtgga tccggttccg
     1141 gagttgcgac agtcattaaa actaaacagg ttgaatttca acttcaggtt ggtgttccac
     1201 tctattttcg gcttcgggca aatccgatca aaactattct cgacaatcaa aagcgcctgg
     1261 acagtaaagg gaatattaaa cgctgtcggg ttccgttaat aaaagaagca gaacaaatcg
     1321 cgtggttgca acgtaaattg ggcaatgcgg cgcgcgttga agatgtgcat cccatatcgg
     1381 aacggccaca gtatttttct ggtgatggta aaagtggaaa gatccaaacg gtttgctttg
     1441 aaggtgtgct caccatcaac gacgcgccag cgttaataga tcttgtacag caaggtattg
     1501 ggccagctaa atcgatggga tgtggcttgc tatctttggc tccactgggc tccggaatgt
     1561 atctcagtaa agtcatcatt gccagggcct ggagcaggga tctttaccaa cttgcccagg
     1621 gattatggca tttatttcca aacagaccgg atgctgctcg tgattttctt tttcatgttg
     1681 agaagcgaaa cacaccagaa ggctgtcatg ttttattgca gtcagcgcaa atgcctgttt
     1741 caactgccgg ttcccgtcgc cgccgtcgtc gccgcggtta agaattctct agagggccct
     1801 attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt
     1861 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     1921 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     1981 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2041 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     2101 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2161 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     2221 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2281 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     2341 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2401 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     2461 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     2521 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     2581 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2641 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2701 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2761 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2821 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2881 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     2941 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     3001 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     3061 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     3121 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     3181 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     3241 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     3301 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     3361 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     3421 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     3481 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     3541 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     3601 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     3661 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     3721 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     3781 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     3841 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     3901 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     3961 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4021 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     4081 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4141 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4201 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4261 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4321 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4381 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4441 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4501 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4561 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4621 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4681 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4741 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4801 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4861 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4921 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4981 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5041 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5101 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5161 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5221 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5281 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5341 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5401 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5461 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5521 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5581 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5641 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5701 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5761 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5821 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5881 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5941 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6001 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6061 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6121 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6181 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6241 acctgacgtc
//
