LOCUS       sequence-170936-        6555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Myc tagged GFP-Vpr fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6555)
  AUTHORS   Votteler J, Ogohara C, Yi S, Hsia Y, Nattermann U, Belnap DM, King 
            NP, Sundquist WI
  TITLE     Designed proteins induce the formation of nanocage-containing 
            extracellular vesicles.
  JOURNAL   Nature. 2016 Dec 8;540(7632):292-295. doi: 10.1038/nature20607. Epub
            2016 Nov 30.
  PUBMED    27919066
REFERENCE   2  (bases 1 to 6555)
  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: 
            "10.1038/nature20607"; journalName: "Nature"; date: "2016-12-8- 8"; 
            volume: "540"; issue: "7632"; pages: "292-295"
COMMENT     Sequence Label: pCMV-Myc-GFP-Vpr-RR
FEATURES             Location/Qualifiers
     source          1..6555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..30
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(31..33,34..744)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(31..33,34..744)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(73..94)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(334..353)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     681..702
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1095..1114)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     primer_bind     complement(1169..1186)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1175..1399
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1445..1873
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1532..1551)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1742..1763
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1882..1902)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1887..2216
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2067..2202
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2129..2148
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2283..3077
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2337..2356)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2947..2966
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3251..3372
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3288..3307)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3342..3361
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3421..3437)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3421..3437)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3434..3456)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3445..3461
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3469..3475,3476..3493,3494..3499))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3514..3535
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3652..3669)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3823..4411)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3903..3922)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4582..5373,5374..5442))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5205..5224
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5443..5547)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5622..5641)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5813..6192
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6193..6396
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6347..6367
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6441..6460
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6441..6459
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gaacagaaac tgattagcga agaagatctg gtgagcaagg gcgaggagct gttcaccggg
       61 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
      121 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
      181 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
      241 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
      301 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
      361 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
      421 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
      481 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
      541 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
      601 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
      661 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
      721 ctcggcatgg acgagctgta caagggaggt ggcggtggag gtgaacaggc accggaagat
      781 caggggcctc agcgtgaacc gcataatgaa tggaccctgg aactgctgga agaactgaaa
      841 cgtgaagcgg tgcgtcattt cccgcgtccg tggctgcatg gcctgggcca gcatatctat
      901 gaaacctatg gcgatacctg ggctggcgtg gaagcgatta ttcgtattct gcagcagctg
      961 ttgtttattc attttcggat aggctgccaa catagccgga ttggcattat tcagcagcgt
     1021 cgtgcgcgtc gtaatggcgc gagccgtagc taagcctagc aaccaacagt atgctcgaga
     1081 agcttgctag cagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     1141 gagcacttaa gtttaaaccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     1201 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     1261 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     1321 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     1381 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     1441 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     1501 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     1561 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     1621 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     1681 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     1741 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     1801 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     1861 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     1921 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     1981 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2041 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2101 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     2161 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     2221 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     2281 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     2341 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     2401 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     2461 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     2521 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     2581 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     2641 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     2701 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     2761 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     2821 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     2881 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     2941 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     3001 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     3061 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     3121 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     3181 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     3241 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3301 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3361 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     3421 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3481 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3541 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3601 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3661 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3721 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3781 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3841 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3901 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3961 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4021 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4081 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4141 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4201 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4261 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4321 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4381 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4441 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4501 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4561 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4621 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4681 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4741 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4801 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4861 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4921 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4981 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5041 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5101 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5161 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5221 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5281 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5341 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5401 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5461 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5521 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     5581 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     5641 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     5701 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     5761 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
     5821 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     5881 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     5941 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     6001 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     6061 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     6121 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     6181 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     6241 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     6301 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     6361 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
     6421 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc tggctagcgt
     6481 ttaaacgggc cctctagaat tatcgatccg gaggtacccg ggcggccgct aggattactg
     6541 ctcggtgaca ccatg
//
