LOCUS       sequence-426582-        7791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces the Lassa virus glycoprotein with a c-terminal HA tag and 
            3xFLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7791)
  AUTHORS   Willard KA, Alston JT, Acciani M, Brindley MA
  TITLE     Identification of Residues in Lassa Virus Glycoprotein Subunit 2 
            That Are Critical for Protein Function.
  JOURNAL   Pathogens. 2018 Dec 26;8(1):1. doi: 10.3390/pathogens8010001.
  PUBMED    30587764
REFERENCE   2  (bases 1 to 7791)
  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: 
            "Pathogens."; date: "2018-12-26"; volume: "8(1):1. doi"; pages: " 
            10.3390/pathogens8010001"
COMMENT     Sequence Label: pcDNAintron-LASV-GPC-HA-c3xFLAG
FEATURES             Location/Qualifiers
     source          1..7791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     572..596
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          696..1268
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     1323..1342
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1323..1341
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1509..1518
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2997..3023
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3033..3098
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3139..3156)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3145..3369
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3415..3843
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3502..3521)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3712..3733
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3852..3872)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3857..4186
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4037..4172
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4099..4118
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4253..5047
                     /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(4307..4326)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4917..4936
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5221..5342
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5258..5277)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5312..5331
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5391..5407)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5391..5407)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5404..5426)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5415..5431
                     /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(5439..5445,5446..5463,5464..5469))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5484..5505
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5622..5639)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5793..6381)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5873..5892)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6552..7343,7344..7412))
                     /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     7175..7194
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7413..7517)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7592..7611)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(7783..7791,1..371)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct
      601 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc
      661 gggggatcga tccccccgat cctgagaact tcagggtgag tttggggacc cttgattgtt
      721 ctttcttttt cgctattgta aaattcatgt tatatggagg gggcaaagtt ttcagggtgt
      781 tgtttagaat gggaagatgt cccttgtatc accatggacc ctcatgataa ttttgtttct
      841 ttcactttct actctgttga caaccattgt ctcctcttat tttcttttca ttttctgtaa
      901 ctttttcgtt aaactttagc ttgcatttgt aacgaatttt taaattcact tttgtttatt
      961 tgtcagattg taagtacttt ctctaatcac ttttttttca aggcaatcag ggtatattat
     1021 attgtacttc agcacagttt tagagaacaa ttgttataat taaatgataa ggtagaatat
     1081 ttctgcatat aaattctggc tggcgtggaa atattcttat tggtagaaac aactacatcc
     1141 tggtcatcat cctgcctttc tctttatggt tacaatgata tacactgttt gagatgagga
     1201 taaaatactc tgagtccaaa ccgggcccct ctgctaacca tgttcatgcc ttcttctttt
     1261 tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat
     1321 tgtaatacga ctcactatag ggcgaattcg gatccactag tccagtgtgg tggaattctg
     1381 cagatctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
     1441 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattcgagc tcatcgatgc
     1501 atggtaccgc caccatgggt cagattgtga cattcttcca ggaagtgcct catgtgattg
     1561 aagaggtgat gaacattgtg ctcattgccc tgtccgtgct cgccgtgctg aaaggtctgt
     1621 acaattttgc cacctgtggc ctggtgggtc tggtgacatt cctcctgctg tgtggtaggt
     1681 cctgcacaac cagcctctac aaaggtgtgt acgagctcca gacactggaa ctgaacatgg
     1741 agacactcaa tatgaccatg cctctctcct gcacaaagaa caacagccat cattacatca
     1801 tggtgggcaa tgagacaggc ctggaactga ccctgaccaa cacaagcatt attaatcaca
     1861 aattttgcaa tctgtccgat gcccacaaaa agaacctcta cgaccacgcc ctcatgagca
     1921 ttatctccac cttccacctg tccatcccca acttcaatca gtacgaggcc atgagctgcg
     1981 attttaatgg cggtaagatt agcgtgcagt acaacctgag ccacagctac gccggcgatg
     2041 ccgccaacca ttgtggtaca gtggccaatg gtgtgctgca gacctttatg aggatggcct
     2101 ggggtggtag ctacattgcc ctcgactccg gcaggggcaa ctgggactgt attatgacca
     2161 gctaccagta cctgattatc cagaatacaa cctgggaaga tcactgccag ttctccagac
     2221 catcccccat cggttacctc ggtctcctct cccagaggac cagagatatt tacattagca
     2281 gaagactgct cggcacattc acatggacac tgtccgattc cgaaggtaaa gacacaccag
     2341 gtggctactg tctgaccagg tggatgctca ttgaggccga actcaaatgc ttcggtaaca
     2401 cagccgtggc caaatgtaat gagaagcatg atgaggaatt ttgtgacatg ctgaggctgt
     2461 ttgacttcaa caaacaggcc attcagaggc tgaaagccga agcccagatg agcattcagc
     2521 tgatcaacaa agccgtgaac gccctgatta atgaccagct catcatgaag aaccatctga
     2581 gggacatcat gggcattcca tactgtaatt acagcaagta ctggtatctc aaccacacaa
     2641 ccacaggtag aacatccctg cccaaatgtt ggctcgtgtc caatggttcc tacctgaacg
     2701 agacccactt ttccgatgat attgaacagc aggccgacaa tatgatcacc gagatgctgc
     2761 agaaggagta catggagagg cagggtaaga caccactggg tctggtggac ctctttgtgt
     2821 tcagcacaag cttctacctc attagcatct tcctccacct ggtgaaaatt ccaacacata
     2881 ggcatattgt gggcaagtcc tgtcccaaac ctcacagact gaatcatatg ggcatttgtt
     2941 cctgtggtct ctacaaacag cctggtgtgc ctgtgaaatg gaagagaggc ggaagctacc
     3001 cgtacgatgt tccagattac gccggagggt ccgactacaa agaccatgac ggtgattata
     3061 aagatcatga tatcgactac aaggatgacg atgacaagtg actcgagtct agagggcccg
     3121 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     3181 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     3241 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     3301 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     3361 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     3421 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3481 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3541 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3601 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3661 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3721 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3781 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3841 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3901 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3961 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4021 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4081 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     4141 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     4201 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     4261 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4321 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4381 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     4441 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4501 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4561 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4621 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4681 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4741 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     4801 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4861 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4921 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4981 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5041 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5101 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5161 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     5221 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5281 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5341 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     5401 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5461 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5521 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5581 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5641 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5701 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5761 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5821 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5881 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5941 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6001 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6061 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6121 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6181 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6241 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6301 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6361 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6421 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6481 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6541 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6601 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6661 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6721 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6781 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6841 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6901 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6961 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7021 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7081 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7141 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7201 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7261 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7321 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7381 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7441 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7501 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     7561 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     7621 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     7681 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
     7741 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga t
//
