LOCUS       sequence-426570-        7782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces the Lassa virus glycoprotein with a c-terminal 3xFLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7782)
  AUTHORS   Acciani M, Alston JT, Zhao G, Reynolds H, Ali AM, Xu B, Brindley MA
  TITLE     Mutational Analysis of Lassa Virus Glycoprotein Highlights Regions 
            Required for Alpha-Dystroglycan Utilization.
  JOURNAL   J Virol. 2017 Aug 24;91(18):e00574-17. doi: 10.1128/JVI.00574-17. 
            Print 2017 Sep 15.
  PUBMED    28679759
REFERENCE   2  (bases 1 to 7782)
  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"; doi: 
            "10.1128/JVI.00574-17"; journalName: "J Virol"; date: "2017-08-24- 
            24"; volume: "91"; issue: "18"; pages: "e00574-17"
COMMENT     Sequence Label: pcDNAintron-LASV-GPC-c3xFLAG
FEATURES             Location/Qualifiers
     source          1..7782
                     /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     815..839
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          939..1511
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     1566..1585
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1566..1584
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1752..1761
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3231..3296
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3373..3390)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3379..3603
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3649..4077
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3736..3755)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3946..3967
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4086..4106)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4091..4420
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4271..4406
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4333..4352
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4487..5281
                     /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(4541..4560)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5151..5170
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5455..5576
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5492..5511)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5546..5565
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5625..5641)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5625..5641)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5638..5660)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5649..5665
                     /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(5673..5679,5680..5697,5698..5703))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5718..5739
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5856..5873)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6027..6615)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6107..6126)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6786..7577,7578..7646))
                     /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     7409..7428
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7647..7751)
                     /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 gcagagctcg tttagtgaac cgtcagatcg
      841 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      901 tccgggggat cgatcccccc gatcctgaga acttcagggt gagtttgggg acccttgatt
      961 gttctttctt tttcgctatt gtaaaattca tgttatatgg agggggcaaa gttttcaggg
     1021 tgttgtttag aatgggaaga tgtcccttgt atcaccatgg accctcatga taattttgtt
     1081 tctttcactt tctactctgt tgacaaccat tgtctcctct tattttcttt tcattttctg
     1141 taactttttc gttaaacttt agcttgcatt tgtaacgaat ttttaaattc acttttgttt
     1201 atttgtcaga ttgtaagtac tttctctaat cacttttttt tcaaggcaat cagggtatat
     1261 tatattgtac ttcagcacag ttttagagaa caattgttat aattaaatga taaggtagaa
     1321 tatttctgca tataaattct ggctggcgtg gaaatattct tattggtaga aacaactaca
     1381 tcctggtcat catcctgcct ttctctttat ggttacaatg atatacactg tttgagatga
     1441 ggataaaata ctctgagtcc aaaccgggcc cctctgctaa ccatgttcat gccttcttct
     1501 ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag
     1561 aattgtaata cgactcacta tagggcgaat tcggatccac tagtccagtg tggtggaatt
     1621 ctgcagatct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
     1681 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattcg agctcatcga
     1741 tgcatggtac cgccaccatg ggtcagattg tgacattctt ccaggaagtg cctcatgtga
     1801 ttgaagaggt gatgaacatt gtgctcattg ccctgtccgt gctcgccgtg ctgaaaggtc
     1861 tgtacaattt tgccacctgt ggcctggtgg gtctggtgac attcctcctg ctgtgtggta
     1921 ggtcctgcac aaccagcctc tacaaaggtg tgtacgagct ccagacactg gaactgaaca
     1981 tggagacact caatatgacc atgcctctct cctgcacaaa gaacaacagc catcattaca
     2041 tcatggtggg caatgagaca ggcctggaac tgaccctgac caacacaagc attattaatc
     2101 acaaattttg caatctgtcc gatgcccaca aaaagaacct ctacgaccac gccctcatga
     2161 gcattatctc caccttccac ctgtccatcc ccaacttcaa tcagtacgag gccatgagct
     2221 gcgattttaa tggcggtaag attagcgtgc agtacaacct gagccacagc tacgccggcg
     2281 atgccgccaa ccattgtggt acagtggcca atggtgtgct gcagaccttt atgaggatgg
     2341 cctggggtgg tagctacatt gccctcgact ccggcagggg caactgggac tgtattatga
     2401 ccagctacca gtacctgatt atccagaata caacctggga agatcactgc cagttctcca
     2461 gaccatcccc catcggttac ctcggtctcc tctcccagag gaccagagat atttacatta
     2521 gcagaagact gctcggcaca ttcacatgga cactgtccga ttccgaaggt aaagacacac
     2581 caggtggcta ctgtctgacc aggtggatgc tcattgaggc cgaactcaaa tgcttcggta
     2641 acacagccgt ggccaaatgt aatgagaagc atgatgagga attttgtgac atgctgaggc
     2701 tgtttgactt caacaaacag gccattcaga ggctgaaagc cgaagcccag atgagcattc
     2761 agctgatcaa caaagccgtg aacgccctga ttaatgacca gctcatcatg aagaaccatc
     2821 tgagggacat catgggcatt ccatactgta attacagcaa gtactggtat ctcaaccaca
     2881 caaccacagg tagaacatcc ctgcccaaat gttggctcgt gtccaatggt tcctacctga
     2941 acgagaccca cttttccgat gatattgaac agcaggccga caatatgatc accgagatgc
     3001 tgcagaagga gtacatggag aggcagggta agacaccact gggtctggtg gacctctttg
     3061 tgttcagcac aagcttctac ctcattagca tcttcctcca cctggtgaaa attccaacac
     3121 ataggcatat tgtgggcaag tcctgtccca aacctcacag actgaatcat atgggcattt
     3181 gttcctgtgg tctctacaaa cagcctggtg tgcctgtgaa atggaagaga gactacaaag
     3241 accatgacgg tgattataaa gatcatgata tcgactacaa ggatgacgat gacaagtgac
     3301 tcgagctagc agatcatcca gcacagtggc ggccgctcga gtctagaggg cccgtttaaa
     3361 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3421 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3481 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3541 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3601 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3661 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3721 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3781 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3841 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3901 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3961 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4021 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     4081 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4141 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4201 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4261 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4321 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     4381 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     4441 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4501 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4561 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4621 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4681 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4741 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4801 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4861 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4921 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4981 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     5041 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5101 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5161 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     5221 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5281 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5341 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5401 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5461 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5521 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5581 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5641 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5701 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5761 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5821 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5881 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5941 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6001 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6061 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6121 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6181 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6241 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6301 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6361 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6421 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6481 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6541 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6601 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6661 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6721 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6781 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6841 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6901 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6961 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7021 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7081 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7141 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7201 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7261 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7321 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7381 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7441 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7501 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7561 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7621 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7681 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7741 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
