LOCUS       sequence-258329-        6792 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Intermediate Filaments, Excitation: 505 / 569, 
            Emission: 516 / 581.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6792)
  AUTHORS   Rizzo MA, Davidson MW, Piston DW
  TITLE     Fluorescent protein tracking and detection: fluorescent protein 
            structure and color variants.
  JOURNAL   Cold Spring Harb Protoc. 2009 Dec;2009(12):pdb.top63. doi: 
            10.1101/pdb.top63.
  PUBMED    20150100
REFERENCE   2  (bases 1 to 6792)
  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: "Cold Spring
            Harb Protoc."; date: "2009-12"; volume: "2009(12):pdb.top63. doi"; 
            pages: " 10.1101/pdb.top63"
COMMENT     Sequence Label: tdEos-Vimentin-N-18
FEATURES             Location/Qualifiers
     source          1..6792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        104..407
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        408..611
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     562..582
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2098..2775
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="MSAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFHGVNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIRMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     CDS             2824..3501
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="SAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVKE
                     GGPLPFAFDILTTAFHYGNRVFAEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIARN
                     DITMEGDTFYNKVRFHGVNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIRMALLLE
                     GNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPDN
                     ARR"
     polyA_signal    3623..3744
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3660..3679)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3714..3733
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3751..4206)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3888..3909)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4100..4119
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4233..4337
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4339..4696
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4339..4530
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4362..4382)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4547..4682
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4731..5525
                     /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             4731..5525
                     /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(4785..4804)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5395..5414
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5713..5732)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5757..5804
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6133..6721
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6622..6641
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
       61 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      121 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      181 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      241 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      301 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      361 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      421 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      481 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      541 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      601 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ccaccatgtc caccaggtcc
      661 gtgtcctcgt cctcctaccg caggatgttc ggcggcccgg gcaccgcgag ccggccgagc
      721 tccagccgga gctacgtgac tacgtccacc cgcacctaca gcctgggcag cgcgctgcgc
      781 cccagcacca gccgcagcct ctacgcctcg tccccgggcg gcgtgtatgc cacgcgctcc
      841 tctgccgtgc gcctgcggag cagcgtgccc ggggtgcggc tcctgcagga ctcggtggac
      901 ttctcgctgg ccgacgccat caacaccgag ttcaagaaca cccgcaccaa cgagaaggtg
      961 gagctgcagg agctgaatga ccgcttcgcc aactacatcg acaaggtgcg cttcctggag
     1021 cagcagaata agatcctgct ggccgagctc gagcagctca agggccaagg caagtcgcgc
     1081 ctgggggacc tctacgagga ggagatgcgg gagctgcgcc ggcaggtgga ccagctaacc
     1141 aacgacaaag cccgcgtcga ggtggagcgc gacaacctgg ccgaggacat catgcgcctc
     1201 cgggagaaat tgcaggagga gatgcttcag agagaggaag ccgaaaacac cctgcaatct
     1261 ttcagacagg atgttgacaa tgcgtctctg gcacgtcttg accttgaacg caaagtggaa
     1321 tctttgcaag aagagattgc ctttttgaag aaactccacg aagaggaaat ccaggagctg
     1381 caggctcaga ttcaggaaca gcatgtccaa atcgatgtgg atgtttccaa gcctgacctc
     1441 acggctgccc tgcgtgacgt acgtcagcaa tatgaaagtg tggctgccaa gaacctgcag
     1501 gaggcagaag aatggtacaa atccaagttt gctgacctct ctgaggctgc caaccggaac
     1561 aatgacgccc tgcgccaggc aaagcaggag tccactgagt accggagaca ggtgcagtcc
     1621 ctcacctgtg aagtggatgc ccttaaagga accaatgagt ccctggaacg ccagatgcgt
     1681 gaaatggaag agaactttgc cgttgaagct gctaactacc aagacactat tggccgcctg
     1741 caggatgaga ttcagaatat gaaggaggaa atggctcgtc accttcgtga ataccaagac
     1801 ctgctcaatg ttaagatggc ccttgacatt gagattgcca cctacaggaa gctgctggaa
     1861 ggcgaggaga gcaggatttc tctgcctctt ccaaactttt cctccctgaa cctgagggaa
     1921 actaatctgg attcactccc tctggttgat acccactcaa aaaggacact tctgattaag
     1981 acggttgaaa ctagagatgg acaggttatc aacgaaactt ctcagcatca cgatgacctt
     2041 gaaagcagcg gtggagcaag cgcagccagt ggtagcgcgg atccaccggt cgccaccatg
     2101 agtgcgatta agccagacat gaagatcaac ctccgtatgg aaggcaacgt aaacgggcac
     2161 cactttgtga tcgacggaga tggtacaggc aagccttttg agggaaaaca gagtatggat
     2221 cttgaagtca aagagggcgg acctctgcct tttgcctttg atatcctgac cactgcattc
     2281 cattacggca acagggtatt cgccgaatat ccagaccaca tacaagacta ttttaagcag
     2341 tcgtttccta aggggtattc gtgggaacga agcttgactt tcgaagacgg gggcatttgc
     2401 attgccagaa acgacataac aatggaaggg gacactttct ataataaagt tcgatttcac
     2461 ggtgtaaact ttcccgccaa tggtccagtt atgcagaaga agacgctgaa atgggagccc
     2521 tccactgaga aaatgtatgt gcgtgatgga gtgctgactg gtgatattcg catggctttg
     2581 ttgcttgaag gaaatgccca ttaccgatgt gacttcagaa ctacttacaa agctaaggag
     2641 aagggtgtca agttaccagg ctaccacttt gtggaccact gcattgagat tttaagccat
     2701 gacaaagatt acaacaaggt taagctgtat gagcatgctg ttgctcattc tggattgcct
     2761 gacaatgcca gacgatccgg tagcggtcac ggtactggtt ctactggttc tggttcttct
     2821 gagagtgcga ttaagccaga catgaagatc aacctccgta tggaaggcaa cgtaaacggg
     2881 caccactttg tgatcgacgg agatggtaca ggcaagcctt ttgagggaaa acagagtatg
     2941 gatcttgaag tcaaagaggg cggacctctg ccttttgcct ttgatatcct gaccactgca
     3001 ttccattacg gcaacagggt attcgccgaa tatccagacc acatacaaga ctattttaag
     3061 cagtcgtttc ctaaggggta ttcgtgggaa cgaagcttga ctttcgaaga cgggggcatt
     3121 tgcattgcca gaaacgacat aacaatggaa ggggacactt tctataataa agttcgattt
     3181 cacggtgtaa actttcccgc caatggtcca gttatgcaga agaagacgct gaaatgggag
     3241 ccctccactg agaaaatgta tgtgcgtgat ggagtgctga ctggtgatat tcgcatggct
     3301 ttgttgcttg aaggaaatgc ccattaccga tgtgacttca gaactactta caaagctaag
     3361 gagaagggtg tcaagttacc aggctaccac tttgtggacc actgcattga gattttaagc
     3421 catgacaaag attacaacaa ggttaagctg tatgagcatg ctgttgctca ttctggattg
     3481 cctgacaatg ccagacgata agcggccgcg actctagatc ataatcagcc ataccacatt
     3541 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     3601 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3661 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3721 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     3781 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     3841 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     3901 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3961 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     4021 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     4081 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     4141 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     4201 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4261 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4321 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     4381 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4441 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4501 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4561 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4621 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4681 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4741 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4801 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4861 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     4921 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4981 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     5041 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5101 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5161 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5221 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     5281 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5341 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5401 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5461 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5521 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5581 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5641 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     5701 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     5761 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     5821 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     5881 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     5941 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     6001 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6061 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6121 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6181 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6241 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6301 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6361 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6421 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6481 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6541 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6601 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6661 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6721 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6781 gttctttcct gc
//
