LOCUS       sequence-74291-e        6111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses UVR8 fused to GFP with a prenylation motif for membrane 
            targeting.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6111)
  AUTHORS   Chen D, Gibson ES, Kennedy MJ
  TITLE     A light-triggered protein secretion system.
  JOURNAL   J Cell Biol. 2013 May 13;201(4):631-40. doi: 10.1083/jcb.201210119.
  PUBMED    23671313
REFERENCE   2  (bases 1 to 6111)
  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: "10.1083/jcb"; 
            journalName: "J Cell Biol"; date: "2013-05-13- 13"; volume: "201"; 
            issue: "4"; pages: "631-40"
COMMENT     Sequence Label: UVR8-GFPCaaX
FEATURES             Location/Qualifiers
     source          1..6111
                     /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      618..627
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1995..2004
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2001..2003,2004..2006,2007..2717)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2001..2003,2004..2006,2007..2717)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2046..2067)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2307..2326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2654..2675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2899..3020
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2936..2955)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2990..3009
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3027..3482)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3164..3185)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3376..3395
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3509..3613
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3615..3972
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3615..3806
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3638..3658)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3823..3958
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4007..4801
                     /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             4007..4801
                     /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(4061..4080)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4671..4690
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4989..5008)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5033..5080
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5409..5997
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5898..5917
                     /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 atctcgagcc accatggcgg aggatatggc tgccgacgaa gttacggctc
      661 ctcctcgtaa ggttcttatc atctccgctg gtgctagcca ctccgtcgct cttctctctg
      721 gtgacattgt ttgttcttgg ggtcgaggag aggatggaca gttaggtcat ggcgatgcag
      781 aggatcgacc ttctccgact cagcttagcg ctttagatgg ccaccaaatt gtttccgtta
      841 cctgtggtgc tgatcacact gttgcttatt cacaatcagg catggaagtc tacagttggg
      901 gatggggtga ttttgggaga ttaggccatg gtaactcaag cgacttgttt actccgctac
      961 caatcaaagc attgcacggt attcggatca agcagattgc ttgtggggat agtcattgtt
     1021 tggctgtcac tatggaagga gaggtccaga gttggggccg caaccagaat ggtcaacttg
     1081 gtctggggga caccgaagat tctctagtgc ctcagaagat tcaagccttt gagggaatac
     1141 gaatcaaaat ggttgctgct ggtgcagaac acactgctgc agttacagaa gatggtgacc
     1201 tctatggatg gggctgggga agatacggaa atttgggatt aggtgaccgg actgaccgct
     1261 tagttcctga aagagttacc tctactggtg gtgagaaaat gtcaatggtt gcttgtggat
     1321 ggcggcacac aatatcagtt tcctactctg gagcattgta tacttatgga tggagcaaat
     1381 atggacagct aggacatgga gacttggagg atcaccttat tcctcacaaa ctggaagcac
     1441 tgagcaacag ttttatctcc cagatttcgg gaggttggag acatacaatg gcattgactt
     1501 cagatggaaa actatatgga tggggttgga ataagtttgg acaagtagga gtcggcaata
     1561 atttagatca gtgttctcct gtgcaagtgc gatttcccga tgatcagaaa gtagttcaag
     1621 tctcatgtgg atggagacat accttggctg tcactgaaag aaataacgtg tttgcttggg
     1681 gtagaggtac aaatggacag ctcggcattg gagagtcggt tgacaggaac tttcccaaga
     1741 ttatagaggc actcagcgtc gatggagcaa gtggacaaca tatagaatct tctaatatcg
     1801 atccatcttc agggaaaagc tgggtgtcgc ctgcagagag atatgcagtt gttcctgatg
     1861 aaacgggcct aacggatggt tcaagcaaag gtaatggagg tgatatcagt gttccacaaa
     1921 ctgatgtcaa gcgtgtacga attaagcttc gaattctgca gtcgacggta ccgcgggccc
     1981 gggatccacc ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     2041 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     2101 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     2161 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     2221 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     2281 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     2341 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     2401 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     2461 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     2521 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2581 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     2641 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2701 tggacgagct gtataagggt aaaaagaaga aaaagaagtc aaagacaaag tgtgtaatta
     2761 tgtagtgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     2821 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     2881 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     2941 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3001 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     3061 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     3121 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     3181 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     3241 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     3301 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     3361 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     3421 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3481 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3541 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3601 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3661 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3721 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3781 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3841 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3901 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3961 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4021 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4081 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4141 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     4201 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4261 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4321 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4381 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4441 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4501 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     4561 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4621 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4681 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4741 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4801 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4861 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4921 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     4981 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     5041 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     5101 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     5161 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     5221 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     5281 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5341 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5401 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5461 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5521 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     5581 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5641 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5701 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5761 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5821 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5881 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5941 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6001 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6061 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
