LOCUS       sequence-420135-        7713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for FLAG-VSVG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7713)
  AUTHORS   Najia MA, Borrajo J, Le A, Tsai F, Huang JY, Griffith LG, Daley GQ, 
            Blainey PC
  TITLE     Live-cell transcriptomics with engineered virus-like particles
  JOURNAL   bioRxiv 2024.10.01.616098
REFERENCE   2  (bases 1 to 7713)
  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 
            2024.10.01.616098"
COMMENT     Sequence Label: pFLAG-VSVG
FEATURES             Location/Qualifiers
     source          1..7713
                     /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"
     CDS             984..1007
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2454..2486
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     CDS             2502..2558
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(3304..3321)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3310..3534
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3580..4008
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3667..3686)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3877..3898
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4017..4037)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4022..4351
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4202..4337
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4264..4283
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4418..5212
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4472..4491)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5082..5101
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5386..5507
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5423..5442)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5477..5496
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5556..5572)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5556..5572)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5569..5591)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5580..5596
                     /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(5604..5610,5611..5628,5629..5634))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5649..5670
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5787..5804)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5958..6546)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6038..6057)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6717..7508,7509..7577))
                     /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     7340..7359
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7578..7682)
                     /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 gctggctagc
      901 ccaccatgaa gtgccttttg tacttagcct ttttattcat tggggtgaat tgcaagttca
      961 ccatagtttt tccatccgga ggagattaca aggatgacga cgataagggc ggaagcttgg
     1021 gacacaacca aaaaggaaac tggaaaaatg ttccttctaa ttaccattat tgcccgtcaa
     1081 gctcagattt aaattggcat aatgacttaa taggcacagc cttacaagtc aaaatgccca
     1141 agagtcacaa ggctattcaa gcagacggtt ggatgtgtca tgcttccaaa tgggtcacta
     1201 cttgtgattt ccgctggtat ggaccgaagt atataacaca ttccatccga tccttcactc
     1261 catctgtaga acaatgcaag gaaagcattg aacaaacgaa acaaggaact tggctgaatc
     1321 caggcttccc tcctcaaagt tgtggatatg caactgtgac ggatgccgaa gcagtgattg
     1381 tccaggtgac tcctcaccat gtgctggttg atgaatacac aggagaatgg gttgattcac
     1441 agttcatcaa cggaaaatgc agcaattaca tatgccccac tgtccataac tctacaacct
     1501 ggcattctga ctataaggtc aaagggctat gtgattctaa cctcatttcc atggacatca
     1561 ccttcttctc agaggacgga gagctatcat ccctgggaaa ggagggcaca gggttcagaa
     1621 gtaactactt tgcttatgaa actggaggca aggcctgcaa aatgcaatac tgcaagcatt
     1681 ggggagtcag actcccatca ggtgtctggt tcgagatggc tgataaggat ctctttgctg
     1741 cagccagatt ccctgaatgc ccagaagggt caagtatctc tgctccatct cagacctcag
     1801 tggatgtaag tctaattcag gacgttgaga ggatcttgga ttattccctc tgccaagaaa
     1861 cctggagcaa aatcagagcg ggtcttccaa tctctccagt ggatctcagc tatcttgctc
     1921 ctaaaaaccc aggaaccggt cctgctttca ccataatcaa tggtacccta aaatactttg
     1981 agaccagata catcagagtc gatattgctg ctccaatcct ctcaagaatg gtcggaatga
     2041 tcagtggaac taccacagaa agggaactgt gggatgactg ggcaccatat gaagacgtgg
     2101 aaattggacc caatggagtt ctgaggacca gttcaggata taagtttcct ttatacatga
     2161 ttggacatgg tatgttggac tccgatcttc atcttagctc aaaggctcag gtgttcgaac
     2221 atcctcacat tcaagacgct gcttcgcaac ttcctgatga tgagagttta ttttttggtg
     2281 atactgggct atccaaaaat ccaatcgagc ttgtagaagg ttggttcagt agttggaaaa
     2341 gctctattgc ctcttttttc tttatcatag ggttaatcat tggactattc ttggttctcc
     2401 gagttggtat ccatctttgc attaaattaa agcacaccaa gaaaagacag atttatacag
     2461 acatagagat gaaccgactt ggaaaggaat tcggatccgg cgcaacaaac ttctctctgc
     2521 tgaaacaagc cggagatgtc gaagagaatc ctggaccggt gtccaagggc gaggaagata
     2581 acatggccat catcaaggag ttcatgaggt ttaaggtcca catggagggt tcagtcaatg
     2641 gccacgagtt cgagattgaa ggcgagggcg agggccgccc ctacgaaggg acacagacgg
     2701 cgaaattgaa ggtgaccaaa ggcgggccat tgcccttcgc atgggacatc ttgtcccctc
     2761 agtttatgta tggcagcaag gcctatgtta agcaccccgc tgatatcccg gactacttga
     2821 agctgtcctt tccagagggg tttaaatggg agcgcgttat gaatttcgaa gacggaggag
     2881 tggttacggt gacgcaggac tcatccctgc aggacggaga atttatatat aaggttaagt
     2941 tgagaggcac aaacttccca agcgacggcc ctgtgatgca gaagaaaaca atggggtggg
     3001 aagcttccag cgagcgcatg taccccgaag atggcgccct caagggcgag ataaagcaaa
     3061 ggctgaaact taaggacggc ggtcattacg acgcggaggt caagacaact tacaaggcta
     3121 aaaaacccgt tcagttgcct ggggcttaca atgttaatat caaacttgac atcacaagcc
     3181 acaatgaaga ctatacgatc gtggagcagt atgaacgagc ggaaggcagg cactcaacgg
     3241 gggggatgga cgagctttac aagtaactcg agtctagagg gcccgtttaa acccgctgat
     3301 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3361 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3421 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3481 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     3541 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3601 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3661 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3721 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3781 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3841 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3901 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3961 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4021 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4081 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4141 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4201 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4261 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     4321 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4381 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4441 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4501 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4561 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4621 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4681 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4741 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4801 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4861 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4921 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4981 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5041 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5101 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5161 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5221 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5281 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5341 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5401 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5461 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5521 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5581 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5641 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5701 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5761 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5821 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5881 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5941 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6001 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6061 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6121 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6181 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6241 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6301 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6361 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6421 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6481 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6541 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6601 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6661 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6721 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6781 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6841 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6901 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6961 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7021 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7081 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7141 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7201 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7261 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7321 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7381 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7441 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7501 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7561 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7621 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7681 cgcacatttc cccgaaaagt gccacctgac gtc
//
