LOCUS Exported 6988 bp ds-DNA circular SYN 05-JAN-2024 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 pcDNA GNSTM-3-RVG-10-Lamp2b-HA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6988) 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 6988) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 5, 2024 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6988 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 110..489 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 490..693 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 644..664 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 738..757 /label=T7 /note="T7 promoter, forward primer" promoter 738..756 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 779..788 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2165..2191 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(2288..2305) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2294..2518 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2564..2992 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2651..2670) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2861..2882 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3001..3021) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3006..3335 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3186..3321 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3248..3267 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3384..4409 /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 4539..4660 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4576..4595) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4630..4649 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4709..4725) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4709..4725) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4722..4744) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4733..4749 /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(4757..4787) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4802..4823 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4940..4957) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5111..5696) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5191..5210) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5867..6727) /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 6490..6509 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6728..6832) /gene="bla" /label=AmpR promoter primer_bind complement(6907..6926) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg 61 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 121 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 181 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 241 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 301 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 361 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 421 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 481 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 541 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 601 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 661 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct 721 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagtgccgc 781 cgccatggtg tgcttccgcc tcttcccggt tccgggctca gggctcgttc tggtctgcct 841 agtcctggga gctgtgcggt cttatgcagg taactcgact atgggcagtg gatacaccat 901 ttggatgccc gagaatccga gaccagggac accttgtgac atttttacca atagcagagg 961 gaagagagca tccaacgggg gcagtggatc tggatccggt ggctcgagtt tggaacttaa 1021 tttgacagat tcagaaaatg ccacttgcct ttatgcaaaa tggcagatga atttcacagt 1081 tcgctatgaa actacaaata aaacttataa aactgtaacc atttcagacc atggcactgt 1141 gacatataat ggaagcattt gtggggatga tcagaatggt cccaaaatag cagtgcagtt 1201 cggacctggc ttttcctgga ttgcgaattt taccaaggca gcatctactt attcaattga 1261 cagcgtctca ttttcctaca acactggtga taacacaaca tttcctgatg ctgaagataa 1321 aggaattctt actgttgatg aacttttggc catcagaatt ccattgaatg acctttttag 1381 atgcaatagt ttatcaactt tggaaaagaa tgatgttgtc caacactact gggatgttct 1441 tgtacaagct tttgtccaaa atggcacagt gagcacaaat gagttcctgt gtgataaaga 1501 caaaacttca acagtggcac ccaccataca caccactgtg ccatctccta ctacaacacc 1561 tactccaaag gaaaaaccag aagctggaac ctattcagtt aataatggca atgatacttg 1621 cctgctggct accatggggc tgcagctgaa catcactcag gataaggttg cttcagttat 1681 taacatcaac cccaatacaa ctcactccac aggcagctgc cgttctcaca ctgctctact 1741 tagactcaat agcagcacta ttaagtatct agactttgtc tttgctgtga aaaatgaaaa 1801 ccgattttat ctgaaggaag tgaacatcag catgtatttg gttaatggct ccgttttcag 1861 cattgcaaat aacaatctca gctactggga tgcccccctg ggaagttctt atatgtgcaa 1921 caaagagcag actgtttcag tgtctggagc atttcagata aatacctttg atctaagggt 1981 tcagcctttc aatgtgacac aaggaaagta ttctacagcc caagagtgtt cgctggatga 2041 tgacaccatt ctaatcccaa ttatagttgg tgctggtctt tcaggcttga ttatcgttat 2101 agtgattgct tacgtaattg gcagaagaaa aagttatgct ggatatcaga ctctgggaag 2161 cggataccca tacgatgtgc cagattacgc ttaaggatcc actagtccag tgtggtggaa 2221 ttctgcagat atccagcaca gtggcggccg ctcgagtcta gagggcccgt ttaaacccgc 2281 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg 2341 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt 2401 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc 2461 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct 2521 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc 2581 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 2641 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 2701 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc 2761 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 2821 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 2881 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt 2941 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt 3001 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 3061 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag 3121 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc 3181 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 3241 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt 3301 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca 3361 ttttcggatc tgatcagcac gtgatgaaaa agcctgaact caccgcgacg tctgtcgaga 3421 agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag 3481 aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct 3541 gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc 3601 cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc 3661 gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc 3721 agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt 3781 tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg 3841 cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt 3901 ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc 3961 acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag 4021 cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct 4081 tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc 4141 atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc 4201 aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat 4261 gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa 4321 gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc 4381 ccagcactcg tccgagggca aaggaatagc acgtgctacg agatttcgat tccaccgccg 4441 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc 4501 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata 4561 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc 4621 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga 4681 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc 4741 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct 4801 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa 4861 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 4921 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 4981 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 5041 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 5101 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 5161 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 5221 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 5281 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 5341 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 5401 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 5461 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 5521 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga 5581 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 5641 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 5701 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 5761 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 5821 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 5881 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 5941 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 6001 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 6061 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 6121 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 6181 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 6241 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 6301 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 6361 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 6421 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 6481 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 6541 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 6601 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 6661 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 6721 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 6781 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 6841 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac 6901 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt 6961 gttggaggtc gctgagtagt gcgcgagc //