LOCUS       sequence-247687-        6949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Golgi/ER Transport, Excitation: 505 / 569, Emission: 
            516 / 581.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6949)
  AUTHORS   Shtengel G, Galbraith JA, Galbraith CG, Lippincott-Schwartz J, 
            Gillette JM, Manley S, Sougrat R, Waterman CM, Kanchanawong P, 
            Davidson MW, Fetter RD, Hess HF
  TITLE     Interferometric fluorescent super-resolution microscopy resolves 3D 
            cellular ultrastructure.
  JOURNAL   Proc Natl Acad Sci U S A. 2009 Mar 3;106(9):3125-30. doi: 
            10.1073/pnas.0813131106. Epub 2009 Feb 6.
  PUBMED    19202073
REFERENCE   2  (bases 1 to 6949)
  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"; doi: 
            "10.1073/pnas.0813131106"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2009-03-3- 3"; volume: "106"; issue: "9"; pages: "3125-30"
COMMENT     Sequence Label: tdEos-VSVG-16
FEATURES             Location/Qualifiers
     source          1..6949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        97..400
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        401..604
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     555..575
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(661..708,709..2193)
                     /codon_start=1
                     /product="vesicular stomatitis virus G glycoprotein, 
                     thermosensitive tsO45 mutant"
                     /label=VSV-G tsO45
                     /note="thermosensitive variant of VSV-G Orsay"
                     /translation="MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSD
                     LNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPS
                     VEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAAIVQVTPHHVLVDEYTGEWVDSQ
                     FINGKCSNDICPTVHNSTTWHSDYKVKGLCDSNLISMDITSFSEDGELSSLGKEGTGFR
                     SNYFAYETGDKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQT
                     SVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPVFTIINGTLK
                     YFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLRTSSGYKFP
                     LYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDETLFFFGDTGLSKNPIEFVEGW
                     FSSWKSSIASFFFIIGLIIGLFLVLRVGIYLCIKLKHTKKRQIYTDIEMNRLG"
     CDS             2248..2925
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="MSAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFHGVNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIRMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     CDS             2974..3651
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="SAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVKE
                     GGPLPFAFDILTTAFHYGNRVFAEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIARN
                     DITMEGDTFYNKVRFHGVNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIRMALLLE
                     GNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPDN
                     ARR"
     polyA_signal    3773..3894
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3810..3829)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3864..3883
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3901..4356)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4038..4059)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4250..4269
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4383..4487
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4489..4846
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4489..4680
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4512..4532)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4697..4832
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4881..5675
                     /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             4881..5675
                     /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(4935..4954)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5545..5564
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5863..5882)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5907..5954
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6283..6871
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6772..6791
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
       61 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      121 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      181 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      241 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      301 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      361 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      421 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      481 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      541 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      601 agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct cgccgccacc
      661 atgaagtgcc ttttgtactt agctttttta ttcatcgggg tgaattgcaa gttcaccata
      721 gtttttccac acaaccaaaa aggaaactgg aaaaatgttc cttccaatta ccattattgc
      781 ccgtcaagct cagatttaaa ttggcataat gacttaatag gcacagcctt acaagtcaaa
      841 atgcccaaga gtcacaaggc tattcaagca gacggttgga tgtgtcatgc ttccaaatgg
      901 gtcactactt gtgatttccg ctggtacgga ccgaagtata taacacattc catccgatcc
      961 ttcactccat ctgtagaaca atgcaaggaa agcattgaac aaacgaaaca aggaacttgg
     1021 ctgaatccag gcttccctcc tcaaagttgt ggatatgcaa ctgtgacgga tgctgaagca
     1081 gcgattgtcc aggtgactcc tcaccatgtg cttgttgatg aatacacagg agaatgggtt
     1141 gattcacagt tcatcaacgg aaaatgcagc aatgacatat gccccactgt ccataactcc
     1201 acaacctggc attccgacta taaggtcaaa gggctatgtg attctaacct catttccatg
     1261 gacatcacct ccttctcaga ggacggagag ctatcatccc taggaaagga gggcacaggg
     1321 ttcagaagta actactttgc ttatgaaacc ggagacaagg cctgcaaaat gcagtactgc
     1381 aagcattggg gagtcagact cccatcaggt gtctggttcg agatggctga taaggatctc
     1441 tttgctgcag ccagattccc tgaatgccca gaagggtcaa gtatctctgc tccatctcag
     1501 acctcagtgg atgtaagtct cattcaggac gttgagagga tcttggatta ttccctctgc
     1561 caagaaacct ggagcaaaat cagagcgggt cttcccatct ctccagtgga tctcagctat
     1621 cttgctccta aaaacccagg aaccggtcct gtctttacca taatcaatgg taccctaaaa
     1681 tactttgaga ccagatacat cagagtcgat attgctgctc caatcctctc aagaatggtc
     1741 ggaatgatca gtggaactac cacagaaagg gaactgtggg atgactgggc tccatatgaa
     1801 gacgtggaaa ttggacccaa tggagttctg aggaccagtt caggatataa gtttccttta
     1861 tatatgattg gacatggtat gttggactcc gatcttcatc ttagctcaaa ggctcaggtg
     1921 tttgaacatc ctcacattca agacgctgct tcgcagcttc ctgatgatga gactttattt
     1981 ttttttggtg atactgggct atccaaaaat ccaatcgagt ttgtagaagg ttggttcagt
     2041 agttggaaga gctctattgc ctcttttttc tttatcatag ggttaatcat tggactattc
     2101 ttggttctcc gagttggtat ttatctttgc attaaattaa agcacaccaa gaaaagacag
     2161 atttatacag acatagagat gaaccgactt ggaaacagaa ttctgcagtc gacggtaccg
     2221 cgggcccggg atccaccggt cgccaccatg agtgcgatta agccagacat gaagatcaac
     2281 ctccgtatgg aaggcaacgt aaacgggcac cactttgtga tcgacggaga tggtacaggc
     2341 aagccttttg agggaaaaca gagtatggat cttgaagtca aagagggcgg acctctgcct
     2401 tttgcctttg atatcctgac cactgcattc cattacggca acagggtatt cgccgaatat
     2461 ccagaccaca tacaagacta ttttaagcag tcgtttccta aggggtattc gtgggaacga
     2521 agcttgactt tcgaagacgg gggcatttgc attgccagaa acgacataac aatggaaggg
     2581 gacactttct ataataaagt tcgatttcac ggtgtaaact ttcccgccaa tggtccagtt
     2641 atgcagaaga agacgctgaa atgggagccc tccactgaga aaatgtatgt gcgtgatgga
     2701 gtgctgactg gtgatattcg catggctttg ttgcttgaag gaaatgccca ttaccgatgt
     2761 gacttcagaa ctacttacaa agctaaggag aagggtgtca agttaccagg ctaccacttt
     2821 gtggaccact gcattgagat tttaagccat gacaaagatt acaacaaggt taagctgtat
     2881 gagcatgctg ttgctcattc tggattgcct gacaatgcca gacgatccgg tagcggtcac
     2941 ggtactggtt ctactggttc tggttcttct gagagtgcga ttaagccaga catgaagatc
     3001 aacctccgta tggaaggcaa cgtaaacggg caccactttg tgatcgacgg agatggtaca
     3061 ggcaagcctt ttgagggaaa acagagtatg gatcttgaag tcaaagaggg cggacctctg
     3121 ccttttgcct ttgatatcct gaccactgca ttccattacg gcaacagggt attcgccgaa
     3181 tatccagacc acatacaaga ctattttaag cagtcgtttc ctaaggggta ttcgtgggaa
     3241 cgaagcttga ctttcgaaga cgggggcatt tgcattgcca gaaacgacat aacaatggaa
     3301 ggggacactt tctataataa agttcgattt cacggtgtaa actttcccgc caatggtcca
     3361 gttatgcaga agaagacgct gaaatgggag ccctccactg agaaaatgta tgtgcgtgat
     3421 ggagtgctga ctggtgatat tcgcatggct ttgttgcttg aaggaaatgc ccattaccga
     3481 tgtgacttca gaactactta caaagctaag gagaagggtg tcaagttacc aggctaccac
     3541 tttgtggacc actgcattga gattttaagc catgacaaag attacaacaa ggttaagctg
     3601 tatgagcatg ctgttgctca ttctggattg cctgacaatg ccagacgata agcggccgcg
     3661 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3721 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3781 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3841 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     3901 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     3961 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     4021 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4081 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     4141 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     4201 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     4261 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     4321 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     4381 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4441 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     4501 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4561 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4621 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4681 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4741 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4801 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     4861 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4921 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4981 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5041 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     5101 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5161 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5221 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5281 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5341 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5401 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5461 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5521 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5581 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5641 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5701 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5761 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5821 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     5881 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     5941 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     6001 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     6061 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     6121 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6181 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6241 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6301 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6361 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6421 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6481 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6541 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6601 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6661 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6721 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6781 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6841 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6901 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatcc
//
