LOCUS       sequence-210879-        6393 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Lysosomes, Excitation: 505 / 569, Emission: 516 / 581.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6393)
  TITLE     Davidson Photoactivatable Fluorescent Proteins
REFERENCE   2  (bases 1 to 6393)
  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"
COMMENT     Sequence Label: tdEos-CathespinB-6
FEATURES             Location/Qualifiers
     source          1..6393
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        115..418
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        419..622
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     573..593
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1710..2387
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="MSAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFHGVNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIRMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     CDS             2436..3113
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="SAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVKE
                     GGPLPFAFDILTTAFHYGNRVFAEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIARN
                     DITMEGDTFYNKVRFHGVNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIRMALLLE
                     GNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPDN
                     ARR"
     polyA_signal    3235..3356
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3272..3291)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3326..3345
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3363..3818)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3500..3521)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3712..3731
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3845..3949
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3951..4308
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3951..4142
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3974..3994)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4159..4294
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4343..5137
                     /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             4343..5137
                     /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(4397..4416)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5007..5026
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5325..5344)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5369..5416
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5745..6333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6234..6253
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
       61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      241 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      421 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      481 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      541 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      601 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggac
      661 tcagatctcg agctcaagct tcgaattcca ggatgtggtg gtccttgatc cctctctctt
      721 gcctgctggc actgaccagt gcccatgaca agccttcctt tcacccactg tcggacgaca
      781 tgattaacta tatcaacaaa cagaatacaa catggcaggc tggacgcaac ttctacaatg
      841 ttgacataag ctatctgaag aagctgtgtg gaactgtcct gggtggaccc aagctgccgg
      901 aaagggttgg gttcagcgag gacataaatc tacctgaatc ctttgatgca cgggaacagt
      961 ggtccaattg cccgaccatc gcacagatca gagaccaggg gtcctgtggc tcttgttggg
     1021 catttggggc agtggaagcc atgtctgacc gaatctgcat tcacaccaat ggccgagtca
     1081 atgtggaggt gtctgctgag gacctgctta cctgctgtgg tatccagtgt ggggatggct
     1141 gtaatggtgg ctatccctct ggagcatgga acttctggac tagaaaaggc ctggtttctg
     1201 gtggagtata caattctcat ataggctgct taccctacac catccctccc tgtgaacacc
     1261 atgtcaatgg ctcccgtccc ccatgcactg gagaaggaga tactcccaag tgcaacaaga
     1321 tgtgtgaggc tggctactcc acatcctaca aggaagataa gcactatggg tacacttcct
     1381 acagtgtgtc tgacagcgag aaggagatca tggcggaaat ctacaaaaat ggcccagtgg
     1441 agggtgcttt tactgtgttt tctgacttct tgacttacaa atcaggcgta tacaagcatg
     1501 aagccggtga tgtgatggga ggccatgcca tccgcattct gggctgggga atagagaatg
     1561 gagtacccta ctggctggta gcaaactcct ggaacgttga ctggggtgat aatggtttct
     1621 ttaaaatcct cagaggagag aaccactgtg gaattgaatc agaaatcgtg gctggaatcc
     1681 cacgcactca ggatccaccg gtcgccacca tgagtgcgat taagccagac atgaagatca
     1741 acctccgtat ggaaggcaac gtaaacgggc accactttgt gatcgacgga gatggtacag
     1801 gcaagccttt tgagggaaaa cagagtatgg atcttgaagt caaagagggc ggacctctgc
     1861 cttttgcctt tgatatcctg accactgcat tccattacgg caacagggta ttcgccgaat
     1921 atccagacca catacaagac tattttaagc agtcgtttcc taaggggtat tcgtgggaac
     1981 gaagcttgac tttcgaagac gggggcattt gcattgccag aaacgacata acaatggaag
     2041 gggacacttt ctataataaa gttcgatttc acggtgtaaa ctttcccgcc aatggtccag
     2101 ttatgcagaa gaagacgctg aaatgggagc cctccactga gaaaatgtat gtgcgtgatg
     2161 gagtgctgac tggtgatatt cgcatggctt tgttgcttga aggaaatgcc cattaccgat
     2221 gtgacttcag aactacttac aaagctaagg agaagggtgt caagttacca ggctaccact
     2281 ttgtggacca ctgcattgag attttaagcc atgacaaaga ttacaacaag gttaagctgt
     2341 atgagcatgc tgttgctcat tctggattgc ctgacaatgc cagacgatcc ggtagcggtc
     2401 acggtactgg ttctactggt tctggttctt ctgagagtgc gattaagcca gacatgaaga
     2461 tcaacctccg tatggaaggc aacgtaaacg ggcaccactt tgtgatcgac ggagatggta
     2521 caggcaagcc ttttgaggga aaacagagta tggatcttga agtcaaagag ggcggacctc
     2581 tgccttttgc ctttgatatc ctgaccactg cattccatta cggcaacagg gtattcgccg
     2641 aatatccaga ccacatacaa gactatttta agcagtcgtt tcctaagggg tattcgtggg
     2701 aacgaagctt gactttcgaa gacgggggca tttgcattgc cagaaacgac ataacaatgg
     2761 aaggggacac tttctataat aaagttcgat ttcacggtgt aaactttccc gccaatggtc
     2821 cagttatgca gaagaagacg ctgaaatggg agccctccac tgagaaaatg tatgtgcgtg
     2881 atggagtgct gactggtgat attcgcatgg ctttgttgct tgaaggaaat gcccattacc
     2941 gatgtgactt cagaactact tacaaagcta aggagaaggg tgtcaagtta ccaggctacc
     3001 actttgtgga ccactgcatt gagattttaa gccatgacaa agattacaac aaggttaagc
     3061 tgtatgagca tgctgttgct cattctggat tgcctgacaa tgccagacga taagcggccg
     3121 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     3181 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3241 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3301 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     3361 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3421 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3481 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3541 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3601 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3661 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3721 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3781 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3841 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3901 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     3961 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4021 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4081 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4141 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4201 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4261 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     4321 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4381 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4441 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4501 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     4561 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4621 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4681 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4741 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4801 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4861 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4921 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4981 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5041 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5101 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     5161 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5221 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5281 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     5341 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     5401 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     5461 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     5521 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     5581 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5641 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5701 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5761 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5821 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5881 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5941 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6001 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6061 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6121 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6181 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6241 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6301 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6361 ttcctggcct tttgctggcc ttttgctcac atg
//
