LOCUS       sequence-426579-        7872 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces the Chikungunya virus nonstructural protein 1 with a 
            c-terminal 3xFLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7872)
  AUTHORS   Reyes Ballista JM, Hoover AJ, Noble JT, Acciani MD, Miazgowicz KL, 
            Harrison SA, Tabscott GAL, Duncan A, Barnes DN, Jimenez AR, Brindley
            MA
  TITLE     Chikungunya virus release is reduced by TIM-1 receptors through 
            binding of envelope phosphatidylserine.
  JOURNAL   J Virol. 2024 Jul 15:e0077524. doi: 10.1128/jvi.00775-24.
  PUBMED    39007616
REFERENCE   2  (bases 1 to 7872)
  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: "J Virol. 
            2024 Jul 15:e0077524. doi: 10.1128/jvi.00775-24."
COMMENT     Sequence Label: pcDNAintron-CHIKV-nsp1-c3xFLAG
FEATURES             Location/Qualifiers
     source          1..7872
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        3..382
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        383..586
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     537..557
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     583..607
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          707..1279
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     1334..1353
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1334..1352
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1520..1529
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3125..3190
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3231..3248)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3237..3461
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3507..3935
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3594..3613)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3804..3825
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3944..3964)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3949..4278
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4129..4264
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4191..4210
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4345..5139
                     /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(4399..4418)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5009..5028
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5313..5434
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5350..5369)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5404..5423
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5483..5499)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5483..5499)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5496..5518)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5507..5523
                     /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(5531..5537,5538..5555,5556..5561))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5576..5597
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5714..5731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5885..6473)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5965..5984)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6644..7435,7436..7504))
                     /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     7267..7286
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7505..7609)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7684..7703)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
       61 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      121 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      181 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      241 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      301 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      361 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      421 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      481 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      541 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg
      601 tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg
      661 atccagcctc cgggggatcg atccccccga tcctgagaac ttcagggtga gtttggggac
      721 ccttgattgt tctttctttt tcgctattgt aaaattcatg ttatatggag ggggcaaagt
      781 tttcagggtg ttgtttagaa tgggaagatg tcccttgtat caccatggac cctcatgata
      841 attttgtttc tttcactttc tactctgttg acaaccattg tctcctctta ttttcttttc
      901 attttctgta actttttcgt taaactttag cttgcatttg taacgaattt ttaaattcac
      961 ttttgtttat ttgtcagatt gtaagtactt tctctaatca cttttttttc aaggcaatca
     1021 gggtatatta tattgtactt cagcacagtt ttagagaaca attgttataa ttaaatgata
     1081 aggtagaata tttctgcata taaattctgg ctggcgtgga aatattctta ttggtagaaa
     1141 caactacatc ctggtcatca tcctgccttt ctctttatgg ttacaatgat atacactgtt
     1201 tgagatgagg ataaaatact ctgagtccaa accgggcccc tctgctaacc atgttcatgc
     1261 cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt
     1321 tggcaaagaa ttgtaatacg actcactata gggcgaattc ggatccacta gtccagtgtg
     1381 gtggaattct gcagatctag agcctctgct aaccatgttc atgccttctt ctttttccta
     1441 cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattcgag
     1501 ctcatcgatg catggtaccg ccaccatgga ttctgtgtac gtggacatag acgctgacag
     1561 cgcctttttg aaggccctgc aacgtgcgta ccccatgttt gaggtggaac ctaggcaggt
     1621 cacatcgaat gaccatgcta atgctagagc gttctcgcat ctagccataa aactaataga
     1681 gcaggaaatt gatcccgact caaccatcct ggatataggt agtgcgccag caaggaggat
     1741 gatgtcggac aggaagtacc actgcgtttg cccgatgcgc agcgcagaag atcccgagag
     1801 actcgctaat tatgcgagaa agctcgcatc tgccgcagga aaagtcctgg acagaaacat
     1861 ttctggaaag atcggggact tacaagcggt gatggccgtg ccagacacgg agacgccaac
     1921 attttgctta cacacagatg tctcatgtag acagagagca gacgtcgcga tataccaaga
     1981 cgtctatgct gtacacgcac ccacgtcgct ataccaccag gcgattaaag gagtccgagt
     2041 ggcgtactgg gtagggttcg acacaacccc gttcatgtac aacgctatgg cgggtgccta
     2101 cccctcatac tcgacaaatt gggcggatga gcaggtactg aaggctaaga acataggatt
     2161 atgttcaaca gacctgacgg aaggtagacg aggcaaattg tctatcatga gagggaaaaa
     2221 gctaaaaccg tgcgaccgtg tgctgttctc agtagggtca acgctttacc cggaaagccg
     2281 cacgctactt aagagctggc acctaccatc ggtgttccat ctaaagggca agcttagctt
     2341 cacatgccgc tgtgacacag tggtttcgtg tgagggctac gtcgttaaga gaataacgat
     2401 gagcccaggc ctttatggaa aaaccatagg gtatgcggta acccaccacg cagacggatt
     2461 cttgatgtgc aagactaccg acacggttga cggcgaaaga gtgtcattct cggtgtgcac
     2521 gtacgtgccg gcgaccattt gtgatcaaat gaccggcatc cttgctacag aagtcacgcc
     2581 ggaggatgca cagaagctgt tggtggggct gaaccagagg atagtggtta acggcagaac
     2641 gcaacggaac acgaacacca tgaagaacta cctacttccc gtggtcgccc aggccttcag
     2701 taagtgggca aaggagtgcc ggaaggacat ggaagatgag aagcttctgg gggtcagaga
     2761 aagaacacta acctgctgct gtctatgggc atttaagaag cagaaaacac acacggtcta
     2821 caagaggcct gatacccagt caatccagaa ggttcaggcc gaatttgaca gctttgtagt
     2881 accgggcctg tggtcgtccg ggttgtcaat cccgttgagg actagaatca agtggttgtt
     2941 acgcaaggtg ccgaaaacag acctgatccc atacagcggg aatgcccaag aagcccagga
     3001 tgcagaaaaa gaagcagagg aagaacgaga agcagaactg actcatgagg ctctaccacc
     3061 cctacaggca gcacaggaag atgtccaggt cgaaatcgac gtggaacagc ttgaggatag
     3121 agcggactac aaagaccatg acggtgatta taaagatcat gatatcgact acaaggatga
     3181 cgatgacaag tgactcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     3241 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3301 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3361 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3421 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3481 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3541 tgtggtggtt acgcgcagcg ttaccgctac acttgccagc gccctagcgc ccgctccttt
     3601 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3661 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3721 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3781 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3841 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3901 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3961 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4021 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4081 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4141 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4201 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4261 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4321 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4381 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4441 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4501 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4561 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4621 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4681 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4741 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4801 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4861 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4921 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4981 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5041 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5101 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5161 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5221 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5281 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5341 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5401 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5461 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5521 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5581 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5641 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5701 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5761 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5821 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5881 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5941 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6001 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6061 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6121 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6181 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6241 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6301 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6361 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6421 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6481 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6541 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6601 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6661 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6721 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6781 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6841 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6901 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6961 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7021 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7081 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7141 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7201 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7261 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7321 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7381 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7441 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7501 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7561 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7621 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     7681 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     7741 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     7801 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     7861 cagatatacg cg
//
