LOCUS       sequence-182520-        5356 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of VAMP3 N-terminally conjugated to FRB and C-terminally 
            conjugated to mCitrine..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5356)
  AUTHORS   Verboogen DRJ, Gonzalez Mancha N, Ter Beest M, van den Bogaart G
  TITLE     Fluorescence lifetime imaging microscopy reveals rerouting of SNARE 
            trafficking driving dendritic cell activation.
  JOURNAL   Elife. 2017 May 19;6. pii: e23525. doi: 10.7554/eLife.23525.
  PUBMED    28524818
REFERENCE   2  (bases 1 to 5356)
  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: "Elife."; 
            date: "2017-05-19"; volume: "6. pii: e23525. doi"; pages: " 
            10.7554/eLife.23525"
COMMENT     Sequence Label: FRB_T2098L-VAMP3-mCherry
FEATURES             Location/Qualifiers
     source          1..5356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1305..1314
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1311..2021
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1456..1474)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1724..1743
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2144..2265
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2181..2200)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2235..2254
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2272..2727)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2409..2430)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2621..2640
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2754..2858
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2860..3217
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2860..3051
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2883..2903)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3068..3203
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3252..4046
                     /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             3252..4046
                     /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(3306..3325)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3916..3935
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4234..4253)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4278..4325
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4654..5242
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5143..5162
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgaggc accatggagc tgatccgcgt ggccatcctg tggcacgaga
      661 tgtggcacga gggcctggag gaggccagcc gcctgtactt cggcgagcgc aacgtgaagg
      721 gcatgttcga ggtgctggag cccctgcacg ccatgatgga gcgcggcccc cagaccctga
      781 aggagaccag cttcaaccag gcctacggcc gcgacctgat ggaggcccag gagtggtgcc
      841 gcaagtacat gaagagcggc aacgtgaagg acctgctgca ggcctgggac ctgtactacc
      901 acgtgttccg ccgcatcagc aagcagtctg gtggcggtgg ctcgggcgga ggtgggtcgg
      961 gtggcggcgg atcagaattc atgtctacag gggtgccttc agggtccagt gctgccactg
     1021 gcagtaatcg aagactccag cagacacaga atcaagtaga tgaggtggtg gacatcatga
     1081 gagtcaatgt ggataaggtg ttagaaagag accagaagct ctcagagcta gacgaccgcg
     1141 cagatgcact gcaggcaggc gcctcgcagt ttgaaacaag tgctgccaag ttgaagagaa
     1201 agtattggtg gaagaactgc aagatgtggg cgatagggat cagtgtcctg gtgattattg
     1261 tcattatcat cattgtgtgg tgtgtctcta aggatccacc ggtcgccacc atggtgagca
     1321 agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg
     1381 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     1441 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg
     1501 acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca
     1561 tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     1621 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca
     1681 tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga
     1741 agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg
     1801 gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga
     1861 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt
     1921 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg
     1981 gccgccactc caccggcggc atggacgagc tgtacaagta aagcggccgc gactctagat
     2041 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     2101 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     2161 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2221 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     2281 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     2341 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     2401 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     2461 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     2521 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     2581 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     2641 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2701 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2761 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2821 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     2881 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2941 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3001 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3061 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3121 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     3181 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     3241 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3301 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3361 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3421 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3481 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3541 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3601 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3661 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3721 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     3781 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3841 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3901 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3961 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4021 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4081 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4141 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4201 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     4261 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     4321 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     4381 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     4441 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     4501 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4561 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4621 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4681 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4741 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4801 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4861 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4921 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4981 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5041 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5101 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5161 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5221 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5281 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5341 tattaccgcc atgcat
//
