LOCUS       sequence-350168-        7580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of lysosome-associated membrane protein 2b 
            fused to the MS2 dimer and C-terminal HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7580)
  AUTHORS   Hung ME, Leonard JN
  TITLE     A platform for actively loading cargo RNA to elucidate limiting 
            steps in EV-mediated delivery.
  JOURNAL   J Extracell Vesicles. 2016 May 13;5:31027. doi: 
            10.3402/jev.v5.31027. eCollection 2016.
  PUBMED    27189348
REFERENCE   2  (bases 1 to 7580)
  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"; journalName: "J Extracell
            Vesicles."; date: "2016-05-13"; volume: "5:31027. doi"; pages: " 
            10.3402/jev.v5.31027. eCollection 2016"
COMMENT     Sequence Label: pMEH60 Lamp2b MS2 HA
FEATURES             Location/Qualifiers
     source          1..7580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      904..913
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2875..2901
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3002..3019)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3008..3232
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3278..3706
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3365..3384)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3575..3596
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3715..3735)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3720..4049
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3900..4035
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3962..3981
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4098..5123
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4098..5123
                     /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    5253..5374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5290..5309)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5344..5363
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5423..5439)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5423..5439)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5436..5458)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5447..5463
                     /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(5471..5477,5478..5495,5496..5501))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5516..5537
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5654..5671)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5825..6413)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5905..5924)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6584..7375,7376..7444))
                     /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     7207..7226
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7445..7549)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 gccgccgcca tggtgtgctt ccgcctcttc ccggttccgg gctcagggct cgttctggtc
      961 tgcctagtcc tgggagctgt gcggtcttat gcattggaac ttaatttgac agattcagaa
     1021 aatgccactt gcctttatgc aaaatggcag atgaatttca cagttcgcta tgaaactaca
     1081 aataaaactt ataaaactgt aaccatttca gaccatggca ctgtgacata taatggaagc
     1141 atttgtgggg atgatcagaa tggtcccaaa atagcagtgc agttcggacc tggcttttcc
     1201 tggattgcga attttaccaa ggcagcatct acttattcaa ttgacagcgt ctcattttcc
     1261 tacaacactg gtgataacac aacatttcct gatgctgaag ataaaggaat tcttactgtt
     1321 gatgaacttt tggccatcag aattccattg aatgaccttt ttagatgcaa tagtttatca
     1381 actttggaaa agaatgatgt tgtccaacac tactgggatg ttcttgtaca agcttttgtc
     1441 caaaatggca cagtgagcac aaatgagttc ctgtgtgata aagacaaaac ttcaacagtg
     1501 gcacccacca tacacaccac tgtgccatct cctactacaa cacctactcc aaaggaaaaa
     1561 ccagaagctg gaacctattc agttaataat ggcaatgata cttgtctgct ggctaccatg
     1621 gggctgcagc tgaacatcac tcaggataag gttgcttcag ttattaacat caaccccaat
     1681 acaactcact ccacaggcag ctgccgttct cacactgctc tacttagact caatagcagc
     1741 accattaagt atctagactt tgtctttgct gtgaaaaatg aaaaccgatt ttatctgaag
     1801 gaagtgaaca tcagcatgta tttggttaat ggctccgttt tcagcattgc aaataacaat
     1861 ctcagctact gggatgcccc cctgggaagt tcttatatgt gcaacaaaga gcaaactgtt
     1921 tcagtgtctg gagcatttca gataaatacc tttgatctaa gggttcagcc tttcaatgtg
     1981 acacaaggaa agtattctac agcccaagag tgttcgctgg atgatgacac cattctaatc
     2041 ccaattatag ttggtgctgg tctttcaggc ttgattatcg ttatagtgat tgcttacgta
     2101 attggcagaa gaaaaagtta tgctggatat cagactctgg gagctagtgg ctctggtagc
     2161 atggcttcta actttactca gttcgttctc gtcgacaatg gcggaactgg cgacgtgact
     2221 gtcgccccaa gcaacttcgc taacgggatc gctgaatgga tcagctctaa ctcgcgttca
     2281 caggcttaca aagtaacctg tagcgttcgt cagagctctg cgcagaatcg caaatacacc
     2341 atcaaagtcg aggtgcctaa aggcgcctgg cgttcgtact taaatatgga actaaccatt
     2401 ccaattttcg ccacgaattc cgactgcgag cttattgtta aggcaatgca aggtctccta
     2461 aaagatggaa acccgattcc ctcagcaatc gcagcgaata gcggtattta tgcaatggcg
     2521 agcaatttca cccagttcgt tctcgtcgac aatggcggaa ctggcgacgt gactgtcgcc
     2581 ccaagcaact tcgctaacgg gatcgctgaa tggatcagct ctaactcgcg ttcacaggct
     2641 tacaaagtaa cctgtagcgt tcgtcagagc tctgcgcaga atcgcaaata caccatcaaa
     2701 gtcgaggtgc ctaaaggcgc ctggcgttcg tacttaaata tggaactaac cattccaatt
     2761 ttcgccacga attccgactg cgagcttatt gttaaggcaa tgcaaggtct cctaaaagat
     2821 ggaaacccga ttccctcagc aatcgcagca aactccggca tctacggaag cggataccca
     2881 tacgatgtgc cagattacgc ttaaggatcg atccactagt ccagtgtggt ggaattctgc
     2941 agatatccag cacagtggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca
     3001 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3061 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3121 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3181 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     3241 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     3301 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3361 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3421 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3481 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3541 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3601 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3661 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3721 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3781 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3841 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3901 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3961 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4021 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4081 gatctgatca gcacgtgatg aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc
     4141 tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc
     4201 gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg
     4261 atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc
     4321 cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg
     4381 cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg
     4441 tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc
     4501 cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg
     4561 ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg
     4621 cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg
     4681 tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca
     4741 ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct
     4801 ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg
     4861 agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct
     4921 atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg
     4981 caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg
     5041 ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca
     5101 ctcgtccgag ggcaaaggaa tagcacgtgc tacgagattt cgattccacc gccgccttct
     5161 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5221 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5281 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5341 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5401 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5461 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5521 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5581 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5641 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5701 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5761 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5821 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5881 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5941 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6001 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6061 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6121 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6181 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6241 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6301 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6361 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6421 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6481 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6541 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6601 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6661 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6721 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6781 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6841 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6901 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6961 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7021 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7081 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7141 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7201 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7261 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7321 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7381 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7441 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7501 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7561 gaaaagtgcc acctgacgtc
//
