LOCUS       sequence-418480-        6991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes (N to C): GNSTM glycosylation motif, 3 residue spacer, RVG 
            peptide, 10 residue spacer, Lamp2b (exosomal transmembrane protein),
            3 residue spacer, HA tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6991)
  AUTHORS   Hung ME, Leonard JN
  TITLE     Stabilization of exosome-targeting peptides via engineered 
            glycosylation.
  JOURNAL   J Biol Chem. 2015 Mar 27;290(13):8166-72. doi: 
            10.1074/jbc.M114.621383. Epub 2015 Feb 5.
  PUBMED    25657008
REFERENCE   2  (bases 1 to 6991)
  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: 
            "10.1074/jbc.M114.621383"; journalName: "J Biol Chem"; date: 
            "2015-03-27- 27"; volume: "290"; issue: "13"; pages: "8166-72"
COMMENT     Sequence Label: pcDNA GNSTM-3-RVG-10-Lamp2b-HA
FEATURES             Location/Qualifiers
     source          1..6991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     562..581
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(800..904)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(979..998)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1170..1549
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1550..1753
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1704..1724
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1798..1817
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1798..1816
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1839..1848
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3225..3251
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3348..3365)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3354..3578
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3624..4052
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3711..3730)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3921..3942
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4061..4081)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4066..4395
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4246..4381
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4308..4327
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4444..5469
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4444..5469
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    5599..5720
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5636..5655)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5690..5709
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5769..5785)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5769..5785)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5782..5804)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5793..5809
                     /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(5817..5823,5824..5841,5842..5847))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5862..5883
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6000..6017)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6171..6759)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6251..6270)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6930..6991,1..730,731..799))
                     /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"
ORIGIN
        1 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
       61 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
      121 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
      181 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
      241 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
      301 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
      361 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
      421 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
      481 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
      541 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
      601 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
      661 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
      721 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
      781 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
      841 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
      901 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
      961 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     1021 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     1081 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
     1141 tcgcgatgta cggaccagat atacgcgttg acattgatta ttgactagtt attaatagta
     1201 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     1261 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     1321 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     1381 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     1441 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     1501 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     1561 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     1621 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     1681 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     1741 tataagcaga gctctctggc taactagaga acccactgct tactggctta tcgaaattaa
     1801 tacgactcac tatagggaga cccaagctgg ctagtgccgc cgccatggtg tgcttccgcc
     1861 tcttcccggt tccgggctca gggctcgttc tggtctgcct agtcctggga gctgtgcggt
     1921 cttatgcagg taactcgact atgggcagtg gatacaccat ttggatgccc gagaatccga
     1981 gaccagggac accttgtgac atttttacca atagcagagg gaagagagca tccaacgggg
     2041 gcagtggatc tggatccggt ggctcgagtt tggaacttaa tttgacagat tcagaaaatg
     2101 ccacttgcct ttatgcaaaa tggcagatga atttcacagt tcgctatgaa actacaaata
     2161 aaacttataa aactgtaacc atttcagacc atggcactgt gacatataat ggaagcattt
     2221 gtggggatga tcagaatggt cccaaaatag cagtgcagtt cggacctggc ttttcctgga
     2281 ttgcgaattt taccaaggca gcatctactt attcaattga cagcgtctca ttttcctaca
     2341 acactggtga taacacaaca tttcctgatg ctgaagataa aggaattctt actgttgatg
     2401 aacttttggc catcagaatt ccattgaatg acctttttag atgcaatagt ttatcaactt
     2461 tggaaaagaa tgatgttgtc caacactact gggatgttct tgtacaagct tttgtccaaa
     2521 atggcacagt gagcacaaat gagttcctgt gtgataaaga caaaacttca acagtggcac
     2581 ccaccataca caccactgtg ccatctccta ctacaacacc tactccaaag gaaaaaccag
     2641 aagctggaac ctattcagtt aataatggca atgatacttg cctgctggct accatggggc
     2701 tgcagctgaa catcactcag gataaggttg cttcagttat taacatcaac cccaatacaa
     2761 ctcactccac aggcagctgc cgttctcaca ctgctctact tagactcaat agcagcacta
     2821 ttaagtatct agactttgtc tttgctgtga aaaatgaaaa ccgattttat ctgaaggaag
     2881 tgaacatcag catgtatttg gttaatggct ccgttttcag cattgcaaat aacaatctca
     2941 gctactggga tgcccccctg ggaagttctt atatgtgcaa caaagagcag actgtttcag
     3001 tgtctggagc atttcagata aatacctttg atctaagggt tcagcctttc aatgtgacac
     3061 aaggaaagta ttctacagcc caagagtgtt cgctggatga tgacaccatt ctaatcccaa
     3121 ttatagttgg tgctggtctt tcaggcttga ttatcgttat agtgattgct tacgtaattg
     3181 gcagaagaaa aagttatgct ggatatcaga ctctgggaag cggataccca tacgatgtgc
     3241 cagattacgc ttaaggatcc actagtccag tgtggtggaa ttctgcagat atccagcaca
     3301 gtggcggccg ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     3361 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3421 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3481 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3541 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3601 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3661 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3721 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3781 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3841 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3901 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3961 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4021 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4081 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4141 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4201 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4261 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4321 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4381 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
     4441 gtgatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc
     4501 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc
     4561 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa
     4621 gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac
     4681 attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg
     4741 ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg
     4801 gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa
     4861 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg
     4921 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat
     4981 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc
     5041 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag
     5101 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg
     5161 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg
     5221 ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt
     5281 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc
     5341 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat
     5401 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca
     5461 aaggaatagc acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5521 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5581 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5641 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5701 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     5761 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5821 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5881 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5941 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6001 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6061 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6121 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6181 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6241 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6301 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6361 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6421 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6481 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6541 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6601 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6661 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6721 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6781 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6841 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6901 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6961 tcagcgatct gtctatttcg ttcatccata g
//
