LOCUS       sequence-478588-        5974 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Contains EGFP-fMCP fluorogenic protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5974)
  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 5974)
  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-fMCP
FEATURES             Location/Qualifiers
     source          1..5974
                     /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"
     promoter        complement(1527..1545)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1528..1545)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1565..1582)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1571..1795
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1841..2269
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1928..1947)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2138..2159
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2278..2298)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2283..2612
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2463..2598
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2525..2544
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2679..3473
                     /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(2733..2752)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3343..3362
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3647..3768
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3684..3703)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3738..3757
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3817..3833)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3817..3833)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3830..3852)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3841..3857
                     /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(3865..3871,3872..3889,3890..3895))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3910..3931
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4048..4065)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4219..4807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4299..4318)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4978..5769,5770..5838))
                     /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     5601..5620
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5839..5943)
                     /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 tgggaggcgg aggctccgga ggcggaggct ccgcctggcg ttcgtactta aatatggaac
     1141 taaccattcc aattttcgcc acgaattccg actgcgagct tattgttaag gcaatgcaag
     1201 gtctcctaaa agatggaaac ccgattccct cagcaatcgc agcaaactcc ggcatctacg
     1261 gctccggaat ggcttctaac tttactcagt tcgttctcgt cgacaatggc ggaactggcg
     1321 acgtgactgt cgccccaagc aacttcgcta acgggatcgc tgaatggatc agctctaact
     1381 cgcgttcaca ggcttacaaa gtaacctgta gcgttcgtca gagctctgcg cagaatcgca
     1441 aatacaccat caaagtcgag gtgcctaaag gcggatcccg tcgccgccgt cgtcgccgcg
     1501 gttaagaatt ctctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga
     1561 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     1621 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     1681 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     1741 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     1801 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     1861 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     1921 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     1981 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2041 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2101 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2161 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2221 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     2281 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2341 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2401 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2461 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2521 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     2581 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     2641 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     2701 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     2761 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     2821 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     2881 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     2941 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3001 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3061 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3121 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3181 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     3241 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3301 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3361 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3421 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     3481 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     3541 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     3601 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     3661 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3721 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     3781 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3841 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3901 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3961 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4021 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4081 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4141 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4201 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4261 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4321 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4381 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4441 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4501 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4561 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4621 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4681 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4741 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4801 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4861 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4921 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4981 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5041 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5101 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5161 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5221 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5281 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5341 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5401 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5461 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5521 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5581 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5641 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5701 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5761 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5821 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5881 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5941 gcgcacattt ccccgaaaag tgccacctga cgtc
//
