LOCUS       sequence-177761-        6287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Membrane/Vesicles, Excitation: 580, Emission: 608.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6287)
  AUTHORS   Shemiakina II, Ermakova GV, Cranfill PJ, Baird MA, Evans RA, 
            Souslova EA, Staroverov DB, Gorokhovatsky AY, Putintseva EV, 
            Gorodnicheva TV, Chepurnykh TV, Strukova L, Lukyanov S, Zaraisky AG,
            Davidson MW, Chudakov DM, Shcherbo D
  TITLE     A monomeric red fluorescent protein with low cytotoxicity.
  JOURNAL   Nat Commun. 2012;3:1204. doi: 10.1038/ncomms2208.
  PUBMED    23149748
REFERENCE   2  (bases 1 to 6287)
  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: "Nat 
            Commun."; date: "2012"; volume: "3:1204. doi"; pages: " 
            10.1038/ncomms2208"
COMMENT     Sequence Label: FusionRed-C-Src-7
FEATURES             Location/Qualifiers
     source          1..6287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             575..616
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     regulatory      2189..2198
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2195..2893
                     /codon_start=1
                     /product="monomeric red fluorescent protein optimized for 
                     use as a fusion tag (Shemiakina et al., 2012)"
                     /label=FusionRed
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMPMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGACDMAL
                     KLVGGGHLICNLETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARY
                     STGGAGDGGK"
     polyA_signal    3015..3136
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3052..3071)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3106..3125
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3143..3598)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3280..3301)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3492..3511
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3625..3729
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3731..4088
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3731..3922
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3754..3774)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3939..4074
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4123..4917
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4123..4917
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4177..4196)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4787..4806
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5105..5124)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5149..5196
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5525..6113
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6014..6033
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcatgggg agcagcaaga gcaagcccaa
      601 ggaccccagc cagcgccggc gcagcctgga gccacccgac agcacccacc acgggggatt
      661 cccagcctcg cagaccccca acaagacagc agcccccgac acgcaccgca cccccagccg
      721 ctcctttggg accgtggcca ccgagcccaa gctcttcggg ggcttcaaca cttctgacac
      781 cgtcacgtcg ccgcagcgtg ccggggcact ggctggcggc gtcaccactt tcgtggctct
      841 ctatgactac gagtcccgga ctgaaacgga cttgtccttc aagaaaggag aacgcctgca
      901 gattgtcaac aacacggaag gtgactggtg gctggctcat tccctcacta caggacagac
      961 gggctacatc cccagtaact atgtcgcgcc ctcagactcc atccaggctg aagagtggta
     1021 ctttgggaag atcactcgtc gggagtccga gcggctgctg ctcaaccccg aaaacccccg
     1081 gggaaccttc ttggtccggg agagcgagac gacaaaaggt gcctattgcc tctccgtttc
     1141 tgactttgac aacgccaagg ggctcaatgt gaagcactac aagatccgca agctggacag
     1201 cggcggcttc tacatcacct cacgcacaca gttcagcagc ctgcagcagc tggtggccta
     1261 ctactccaaa catgctgatg gcttgtgcca ccgcctgacc aacgtctgcc ccacgtccaa
     1321 gccccagacc cagggactcg ccaaggacgc gtgggaaatc ccccgggagt cgctgcggct
     1381 ggaggtgaag ctggggcagg gctgctttgg agaggtctgg atggggacct ggaacggcac
     1441 caccagagtg gccataaaga ctctgaagcc cggcaccatg tccccggagg ccttcctgca
     1501 ggaagcccaa gtgatgaaga agctccggca tgagaagctg gttcagctgt acgcagtggt
     1561 gtcggaagag cccatctaca tcgtcactga gtacatgagc aaggggagcc tcctggattt
     1621 cctgaaggga gagatgggca agtacctgcg gctgccacag ctcgtcgata tggctgctca
     1681 gattgcatcc ggcatggcct atgtggagag gatgaactac gtgcaccgag acctgcgggc
     1741 ggccaacatc ctggtggggg agaacctggt gtgcaaggtg gctgactttg ggctggcacg
     1801 cctcatcgag gacaacgagt acacagcacg gcaaggtgcc aagttcccca tcaagtggac
     1861 agcccccgag gcagccctct atggccggtt caccatcaag tcggatgtct ggtccttcgg
     1921 catcctgctg actgagctga ccaccaaggg ccgggtgcca tacccaggga tggtcaacag
     1981 ggaggtgctg gaccaggtgg agaggggcta ccgcatgccc tgcccgcccg agtgccccga
     2041 gtcgctgcat gacctcatgt gccagtgctg gcggaaggac cctgaggagc ggcccacttt
     2101 tgagtacctg caggccttcc tggaggacta cttcacctcg acagagcccc agtaccagcc
     2161 tggagagaac ctacgggatc caccggtcgc caccatggtg agcgagctga ttaaggagaa
     2221 catgcccatg aagctgtaca tggagggcac cgtgaacaac caccacttca agtgcacatc
     2281 cgagggcgaa ggcaagccct acgagggcac ccagaccatg agaatcaagg tcgtcgaggg
     2341 cggccctctc cccttcgcct tcgacatcct ggctaccagc ttcatgtacg gcagcagaac
     2401 cttcatcaag caccctccgg gcatccccga cttctttaag cagtccttcc ctgagggctt
     2461 cacatgggag agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac
     2521 cagcctccag gacggctgcc tcatctacaa cgtcaaggtt agaggggtga acttcccagc
     2581 caacggccct gtgatgcaga agaaaacact cggctgggag gcctccaccg agacgatgta
     2641 ccccgctgac ggcggcctgg aaggcgcatg tgacatggcc ctgaagctcg tgggcggggg
     2701 ccacctgatc tgcaaccttg agaccacata cagatccaag aaacccgcta cgaacctcaa
     2761 gatgcccggc gtctacaacg tggaccacag actggaaaga atcaaggagg ccgacgatga
     2821 gacctacgtc gagcagcacg aggtggctgt ggccagatac tctactggtg gcgctggtga
     2881 tggaggtaaa tgagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     2941 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     3001 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3061 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3121 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     3181 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3241 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3301 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3361 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3421 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3481 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     3541 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     3601 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3661 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3721 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     3781 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3841 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3901 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3961 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4021 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4081 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4141 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4201 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4261 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     4321 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4381 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4441 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4501 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4561 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4621 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     4681 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4741 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4801 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4861 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4921 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4981 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5041 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     5101 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     5161 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     5221 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     5281 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     5341 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     5401 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5461 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5521 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5581 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5641 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5701 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5761 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5821 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5881 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5941 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6001 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6061 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6121 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6181 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag
     6241 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccg
//
