LOCUS       sequence-256755-        4941 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Ras GTPase, Excitation: 448 / 473, Emission: 500 / 
            528.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4941)
  AUTHORS   Shu X, Lev-Ram V, Deerinck TJ, Qi Y, Ramko EB, Davidson MW, Jin Y, 
            Ellisman MH, Tsien RY
  TITLE     A genetically encoded tag for correlated light and electron 
            microscopy of intact cells, tissues, and organisms.
  JOURNAL   PLoS Biol. 2011 Apr;9(4):e1001041. doi: 
            10.1371/journal.pbio.1001041. Epub 2011 Apr 5.
  PUBMED    21483721
REFERENCE   2  (bases 1 to 4941)
  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: "PLoS 
            Biol."; date: "2011-04"; volume: "9(4):e1001041. doi"; pages: " 
            10.1371/journal.pbio.1001041. Epub 2011 Apr 5"
COMMENT     Sequence Label: miniSOG-Rab5a-7
FEATURES             Location/Qualifiers
     source          1..4941
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        63..366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        367..570
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     521..541
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      609..618
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             615..932
                     /codon_start=1
                     /product="mini singlet oxygen generator, a fluorescent 
                     flavoprotein suitable for correlated light and electron 
                     microscopy (Shu et al., 2011)"
                     /label=miniSOG
                     /translation="MEKSFVITDPRLPDNPIIFASDGFLELTEYSREEILGRNGRFLQG
                     PETDQATVQKIRDAIRDQREITVQLINYTKSGKKFWNLLHLQPMRDQKGELQYFIGVQL
                     DG"
     polyA_signal    1731..1852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1768..1787)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1822..1841
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1859..2314)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1996..2017)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2208..2227
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2341..2445
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2447..2804
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2447..2638
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2470..2490)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2655..2790
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2839..3633
                     /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             2839..3633
                     /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(2893..2912)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3503..3522
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3821..3840)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3865..3912
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4241..4829
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4730..4749
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      541 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc
      601 taccggtcgc caccatggag aaaagtttcg tgataactga tccacggctg ccagacaatc
      661 ccatcatctt cgcatccgat ggcttcctgg agctgaccga gtattccaga gaggagatcc
      721 tgggccgcaa tggccgcttt ctgcagggac cagagacaga ccaggccaca gtgcagaaga
      781 ttcgcgatgc cattagagat cagcgcgaga ttaccgtgca gctgataaac tacacaaaaa
      841 gcgggaagaa attctggaac ctcctgcacc tccagcccat gagggaccag aagggtgagc
      901 tccagtattt catcggagtg cagctggatg gatccggact cagatctcga gccatggcta
      961 gtcgaggcgc aacaagaccc aacgggccaa atactggaaa taaaatatgc cagttcaaac
     1021 tagtacttct gggagagtct gctgttggca aatcaagcct agtgcttcgt tttgtgaagg
     1081 gccaatttca tgaatttcaa gagagtacca taggggctgc ttttctaacc caaactgtgt
     1141 gtcttgatga tacaacagta aagtttgaaa tatgggatac agctggtcaa gaacgatacc
     1201 atagcttagc accaatgtac tacagaggag cacaagcagc catagttgta tatgatatca
     1261 caaatgagga gtcctttgcc agagccaaaa actgggttaa agaacttcag aggcaagcca
     1321 gtcctaacat tgtaatagct ttatcaggaa acaaggctga tcttgcaaat aaaagagctg
     1381 tcgatttcca ggaagcacag tcctatgcag atgacaacag tttattattc atggagacat
     1441 cagctaaaac atcgatgaat gtaaatgaaa tattcatggc aatagctaaa aagttgccaa
     1501 agaacgaacc acagaatcca ggagcaaatt ctgccagagg aagaggagta gaccttactg
     1561 aacccacgca gccaaccagg aatcagtgtt gtagtaacta aggatccacc ggatctagat
     1621 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     1681 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     1741 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     1801 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
     1861 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     1921 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     1981 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     2041 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     2101 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     2161 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     2221 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     2281 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     2341 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2401 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     2461 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     2521 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     2581 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     2641 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     2701 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     2761 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     2821 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     2881 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     2941 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3001 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     3061 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3121 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3181 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3241 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3301 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3361 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3421 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3481 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     3541 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     3601 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     3661 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     3721 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     3781 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     3841 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     3901 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     3961 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     4021 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     4081 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4141 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4201 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4261 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4321 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4381 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4441 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4501 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4561 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4621 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4681 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4741 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4801 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4861 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     4921 ataaccgtat taccgccatg c
//
