LOCUS       sequence-170937-        6229 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Myc tagged GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6229)
  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 6229)
  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
FEATURES             Location/Qualifiers
     source          1..6229
                     /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..747)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(31..33,34..744)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /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(769..788)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     primer_bind     complement(843..860)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    849..1073
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1119..1547
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1206..1225)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1416..1437
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1556..1576)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1561..1890
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1741..1876
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1803..1822
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1957..2751
                     /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(2011..2030)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2621..2640
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2925..3046
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2962..2981)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3016..3035
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3095..3111)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3095..3111)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3108..3130)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3119..3135
                     /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(3143..3149,3150..3167,3168..3173))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3188..3209
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3326..3343)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3497..4085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3577..3596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4256..5047,5048..5116))
                     /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     4879..4898
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5117..5221)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5296..5315)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5487..5866
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5867..6070
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6021..6041
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6115..6134
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6115..6133
                     /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 caagtaactc gagaagcttg ctagcagatc tttttccctc
      781 tgccaaaaat tatggggaca tcatgaagcc ccttgagcac ttaagtttaa accgctgatc
      841 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
      901 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
      961 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     1021 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     1081 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     1141 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     1201 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     1261 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     1321 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     1381 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     1441 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     1501 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     1561 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     1621 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     1681 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     1741 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     1801 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     1861 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     1921 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     1981 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     2041 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     2101 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     2161 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     2221 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     2281 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     2341 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     2401 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     2461 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     2521 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     2581 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     2641 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     2701 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     2761 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     2821 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     2881 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     2941 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3001 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     3061 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     3121 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     3181 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     3241 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     3301 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3361 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3421 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3481 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3541 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3601 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3661 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3721 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3781 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3841 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3901 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3961 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4021 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4081 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4141 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4201 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4261 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4321 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4381 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4441 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4501 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4561 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4621 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4681 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4741 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4801 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4861 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4921 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4981 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5041 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5101 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5161 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5221 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     5281 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     5341 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     5401 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     5461 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
     5521 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     5581 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     5641 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     5701 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     5761 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     5821 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     5881 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     5941 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     6001 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     6061 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
     6121 gactcactat agggagaccc aagctggcta gcgtttaaac gggccctcta gaattatcga
     6181 tccggaggta cccgggcggc cgctaggatt actgctcggt gacaccatg
//
