LOCUS       sequence-468750-        5079 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For production of Extracellular Vesicles (EVs), with the 
            transmembrane region of Neuraminidase protein fused to Streptavidin 
            on the vesicles surface.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5079)
  AUTHORS   Gonzalez-Magaldi M, Gullapalli A, Papoulas O, Liu C, Leung AY, Guo 
            L, Brilot A, Marcotte EM, Ke Z, Leahy DJ
  TITLE     Structure and organization of full-length Epidermal Growth Factor 
            Receptor in extracellular vesicles by cryo-electron tomography.
  JOURNAL   bioRxiv [Preprint]. 2024 Nov 28:2024.11.25.625301. doi: 
            10.1101/2024.11.25.625301.
  PUBMED    39651119
REFERENCE   2  (bases 1 to 5079)
  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: "bioRxiv 
            [Preprint]. 2024 Nov 28:2024.11.25.625301. doi: 
            10.1101/2024.11.25.625301."
COMMENT     Sequence Label: pEABR NA 3C FLAG SA
FEATURES             Location/Qualifiers
     source          1..5079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        96..475
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        476..679
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     630..650
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     676..700
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1262..1285
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1286..1357
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     primer_bind     complement(1875..1892)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1881..2105
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2179..2198)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2180..2198)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2205..2222)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2205..2221)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2214..2236)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2363..2818
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2450..2469)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2660..2681
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2844..2948
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2949..3017,3018..3809)
                     /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     complement(3167..3186)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3980..4568
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4469..4488
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4722..4739
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4856..4877
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4892..4897,4898..4915,4916..4922)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4930..4946
                     /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)."
     primer_bind     4935..4957
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4954..4970
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4954..4970
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4991..5009
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ataaatcaat attggctatt ggccattgca tacgttgtat ccatatcata atatgtacat
       61 ttatattggc tcatgtccaa cattaccgcc atgttgacat tgattattga ctagttatta
      121 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      181 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      241 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      301 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      361 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      421 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      481 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      541 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      601 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      661 ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac gccatccacg
      721 ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccgcggcc ccaagcttgc
      781 tttgcctggc aatcccgatc acagggaaat gggggaaact ctcccagaag aggttggaga
      841 atatcggcag ccatctggtg gcagtgtacc tgtgtctccg ggtccgccca gtggtcttga
      901 acccactagc tcctcacctt acggcggcgg ctcctttaac agctcaatca acaacatcca
      961 cgaaatggag attcagctga aggatgcact cgaaaagaat caacaatggc tcgtttacga
     1021 ccaacaacgc gaggtgtacg tgaaaggatt gcttgcaaaa attttcgaac tggaaaagaa
     1081 aacagagact gcagcacata gtcttcctat gaacccgaac cagaaaatta ttacgattgg
     1141 atctatctgc cttgtagtcg gtctcatcag cctgatcctc caaataggga atataatctc
     1201 aatatggatc agtcactcaa tacagaccgg cagtcagaat cacaccggag gcggcggatc
     1261 ccttgaagtc ctctttcagg gacccgatta taaggacgat gacgataagg attacaagga
     1321 tgatgatgac aaggattaca aggacgacga tgacaaggga ggcggatccg acccatccaa
     1381 ggactccaaa gcacaagtgt ctgcggctga agccgggata actgggacgt ggtacaacca
     1441 acttgggtct acgtttatag tcactgctgg ggcagacgga gcccttactg gtacctatgt
     1501 gaccgcgaga ggaaacgcag aaagccgcta cgttctcaca ggtcggtatg atagcgcccc
     1561 tgcgacagat gggagcggaa cagcattggg ttggaccgtc gcgtggaaga ataactacag
     1621 aaacgctcat agtgctacca cctggtctgg acagtatgta ggtggagcgg aggcccgaat
     1681 aaacacccag tggctcctca cgtcaggtac aacggaggca aacgcgtgga aatctacact
     1741 tgttggtcac gacacgttca ccaaagtcaa accgtcagca gcaagtatcg atgctgcaaa
     1801 gaaggctggg gtgaacaatg gtaatccgct tgacgcagta cagcagtgag aattcctgca
     1861 gccccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     1921 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     1981 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2041 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     2101 tatggcttct gaggcggaaa gaaccagctg gggctcgatc ctctagagcg gccgccactg
     2161 cggtggagct ccaattcgcc ctatagtgag tcgtattaca attcactggc cgtcgtttta
     2221 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     2281 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     2341 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2401 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2461 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2521 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2581 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2641 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2701 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2761 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     2821 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2881 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2941 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3001 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3061 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3121 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3181 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3241 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3301 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3361 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3421 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3481 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3541 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3601 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3661 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3721 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3781 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     3841 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     3901 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3961 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4021 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4081 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4141 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4201 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4261 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4321 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4381 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4441 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4501 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4561 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4621 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     4681 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     4741 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     4801 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     4861 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     4921 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     4981 ccaagctcga aattaaccct cactaaaggg aacaaaagct ggtaccgggc cccccctcga
     5041 gatccggcca ttagccatat tattcattgg ttatatagc
//
