LOCUS       sequence-478930-        5322 bp    DNA     circular SYN 17-JUL-2025
DEFINITION  Localization: Ras GTPase, Excitation: 490 / 553, Emission: 507 / 
            573.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5322)
  TITLE     Davidson Photoactivatable Fluorescent Proteins
REFERENCE   2  (bases 1 to 5322)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: Dendra2-Rab4a-7
FEATURES             Location/Qualifiers
     source          1..5322
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(105..693)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(185..204)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    1022..1069
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1094..1113
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1301..2095)
                     /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             complement(1301..2095)
                     /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(1412..1431)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2022..2041
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2130..2487)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2144..2279
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        2296..2487
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2444..2464
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(2489..2593)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2620..3075
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2707..2726)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2917..2938
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    3082..3203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3093..3112)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3147..3166
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4011..4700)
                     /codon_start=1
                     /product="monomeric photoswitchable green-to-red 
                     fluorescent protein from Dendronephthya"
                     /label=Dendra2
                     /note="mammalian codon-optimized"
                     /translation="MNTPGINLIKEDMRVKVHMEGNVNGHAFVIEGEGKGKPYEGTQTA
                     NLTVKEGAPLPFSYDILTTAVHYGNRVFTKYPEDIPDYFKQSFPEGYSWERTMTFEDKG
                     ICTIRSDISLEGDCFFQNVRFKGTNFPPNGPVMQKKTLKWEPSTEKLHVRDGLLVGNIN
                     MALLLEGGGHYLCDFKTTYKAKKVVQLPDAHFVDHRIEILGNDSDYNKVKLYEHAVARY
                     SPLPSQVW"
     promoter        complement(4745..4948)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4774..4794)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4949..5252
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
       61 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
      121 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
      181 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
      241 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
      301 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
      361 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
      421 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
      481 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
      541 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
      601 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
      661 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
      721 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
      781 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
      841 tatgagtaac ctgaggctat ggcagggcct gccgccccga cgttggctgc gagccctggg
      901 ccttcacccg aacttggggg gtggggtggg gaaaaggaag aaacgcgggc gtattggccc
      961 caatggggtc tcggtggggt atcgacagag tgccagccct gggaccgaac cccgcgttta
     1021 tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg
     1081 gttccttccg gtattgtctc cttccgtgtt tcagttagcc tccccctagg gtgggcgaag
     1141 aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt
     1201 ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg
     1261 ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa
     1321 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc
     1381 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct
     1441 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt
     1501 ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg
     1561 gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt
     1621 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat
     1681 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg
     1741 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc
     1801 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag
     1861 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttgcagtt
     1921 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca
     1981 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata
     2041 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa
     2101 acgatcctca tcctgtctct tgatcgatct ttgcaaaagc ctaggcctcc aaaaaagcct
     2161 cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa
     2221 aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg
     2281 ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt
     2341 ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg
     2401 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca
     2461 ttccacagct ggttctttcc gcctcaggac tcttcctttt tcaatattat tgaagcattt
     2521 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2581 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     2641 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2701 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2761 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2821 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2881 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2941 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3001 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3061 taacgcttac aatttacgcg ttaagataca ttgatgagtt tggacaaacc acaactagaa
     3121 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     3181 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     3241 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg
     3301 attatgatca gttatctaga tccggtggat ccctaacaac cacactcctg agcgttcggg
     3361 gcctgtgcgc gccgcggtga cctcagctgt ctcaaggcag catctccgta ctgaatacct
     3421 gagcccattc tttctgggtc cagctcacct gattcgattt tgttaagtat ttttcttgca
     3481 cactgtacaa aagcctcttc tacattctcc cctgtgagcg cacttgtttc caaaaacatc
     3541 agctcatttt cttgagcaaa tctggaggct tctaagaagg taacttcacg atctgcatcc
     3601 aggtccttct tgtttccaca aaggatgatc acaatgttct ggctcgccag cattcgggca
     3661 tctgttaacc aattagtaag cgcattgtag gtttctcggc tggtgatatc atagacgagg
     3721 agagccccgg ccgcgcctcg gtaataactt ctcgtcacgg acctgaatcg ttcttgtcct
     3781 gctgtatccc atatttgtaa ctttacatat ttaccaccaa catttattat ctttgaacca
     3841 aattccactc ctattgtatg atttgagtca tctttgaatt ttttttcaat aaactgatga
     3901 agtaagcaag atttgccagt tcctgcattt ccaataacca agaacttaaa caaaaaatcg
     3961 taggtttcgg acatggccgt ctgcgacatg gctcgagatc tgagtccgga ccacacctgg
     4021 ctgggcaggg ggctgtagcg ggccacggcg tgctcgtaca gcttcacctt gttgtagtcg
     4081 ctgtcgttgc ccaggatctc gatgcggtgg tccacgaagt gggcgtcggg cagctgcacc
     4141 accttcttgg ccttgtaggt ggtcttgaag tcgcacaggt agtggccgcc gccctccagc
     4201 agcagggcca tgttgatgtt gcccaccagc aggccgtcgc gcacgtgcag cttctcggtg
     4261 ctgggctccc acttcagggt cttcttctgc atcacggggc cgttgggggg gaagttggtg
     4321 cccttgaagc gcacgttctg gaagaagcag tcgccctcca ggctgatgtc gctgcggatg
     4381 gtgcagatgc ccttgtcctc gaaggtcatg gtgcgctccc agctgtagcc ctcggggaag
     4441 ctctgcttga agtagtcggg gatgtcctcg gggtacttgg tgaacacccg gttgccgtag
     4501 tgcacggcgg tggtcaggat gtcgtagctg aagggcaggg gggcgccctc cttcacggtc
     4561 aggttggcgg tctgggtgcc ctcgtagggc ttgcccttgc cctcgccctc gatcacgaag
     4621 gcgtggccgt tcacgttgcc ctccatgtgc accttcacgc gcatgtcctc cttgatcagg
     4681 ttaattcccg gggtgttcat ggtggcgacc ggtagcgcta gcggatctga cggttcacta
     4741 aaccagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
     4801 gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc
     4861 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
     4921 cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt
     4981 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     5041 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     5101 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     5161 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     5221 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     5281 atgaccccgt aattgattac tattaataac taatgcatgg cg
//
