LOCUS       sequence-466801-        5634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For production of Viral like particles (VLPs) by expressing the MLV 
            GAG protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5634)
  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 5634)
  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: pBS CMV GAG
FEATURES             Location/Qualifiers
     source          1..5634
                     /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             837..1253
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1183..1205
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1221..1237
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     polyA_signal    2436..2660
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2734..2753)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2735..2753)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2760..2777)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2760..2776)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2769..2791)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2918..3373
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3005..3024)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3215..3236
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3399..3503
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3504..3572,3573..4364)
                     /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(3722..3741)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4535..5123
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5024..5043
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5277..5294
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5411..5432
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5447..5452,5453..5470,5471..5477)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5485..5501
                     /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     5490..5512
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5509..5525
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5509..5525
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5546..5564
                     /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 ccaagcttgg
      781 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aatatgggcc
      841 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
      901 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
      961 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
     1021 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
     1081 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
     1141 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
     1201 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gccaaaccta
     1261 aacctcaagt tctttctgac agtggggggc cgctcatcga cctacttaca gaagaccccc
     1321 cgccttatag ggacccaaga ccaccccctt ccgacaggga cggaaatggt ggagaagcga
     1381 cccctgcggg agaggcaccg gacccctccc caatggcatc tcgcctacgt gggagacggg
     1441 agccccctgt ggccgactcc actacctcgc aggcattccc cctccgcgca ggaggaaacg
     1501 gacagcttca atactggccg ttctcctctt ctgaccttta caactggaaa aataataacc
     1561 cttctttttc tgaagatcca ggtaaactga cagctctgat cgagtctgtc ctcatcaccc
     1621 atcagcccac ctgggacgac tgtcagcagc tgttggggac tctgctgacc ggagaagaaa
     1681 aacaacgggt gctcttagag gctagaaagg cggtgcgggg cgatgatggg cgccccactc
     1741 aactgcccaa tgaagtcgat gccgcttttc ccctcgagcg cccagactgg gattacacca
     1801 cccaggcagg taggaaccac ctagtccact atcgccagtt gctcctagcg ggtctccaaa
     1861 acgcgggcag aagccccacc aatttggcca aggtaaaagg aataacacaa gggcccaatg
     1921 agtctccctc ggccttccta gagagactta aggaagccta tcgcaggtac actccttatg
     1981 accctgagga cccagggcaa gaaactaatg tgtctatgtc tttcatttgg cagtctgccc
     2041 cagacattgg gagaaagtta gagaggttag aagatttaaa aaacaagacg cttggagatt
     2101 tggttagaga ggcagaaaag atctttaata aacgagaaac cccggaagaa agagaggaac
     2161 gtatcaggag agaaacagag gaaaaagaag aacgccgtag gacagaggat gagcagaaag
     2221 agaaagaaag agatcgtagg agacatagag agatgagcaa gctattggcc actgtcgtta
     2281 gtggacagaa acaggataga cagggaggag aacgaaggag gtcccaactc gatcgcgacc
     2341 agtgtgccta ctgcaaagaa aaggggcact gggctaaaga ttgtcccaag aaaccacgag
     2401 gacctcgggg accaagaccc cagacctccc tcctgctgtg ccttctagtt gccagccatc
     2461 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2521 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2581 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     2641 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct cgatcctcta
     2701 gagcggccgc caccgcggtg gagctccaat tcgccctata gtgagtcgta ttacaattca
     2761 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     2821 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     2881 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     2941 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3001 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3061 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3121 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3181 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3241 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3301 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     3361 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3421 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3481 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3541 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3601 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3661 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3721 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3781 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3841 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3901 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3961 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4021 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4081 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4141 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4201 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4261 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4321 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4381 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4441 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4501 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4561 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4621 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4681 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4741 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4801 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4861 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4921 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4981 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5041 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5101 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5161 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5221 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     5281 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     5341 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     5401 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     5461 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     5521 atgaccatga ttacgccaag ctcgaaatta accctcacta aagggaacaa aagctggtac
     5581 cgggcccccc ctcgagatcc ggccattagc catattattc attggttata tagc
//
