LOCUS       sequence-229588-        6042 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Ectopic expression of ZIKV protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6042)
  AUTHORS   Hertzog J, Dias Junior AG, Rigby RE, Donald CL, Mayer A, Sezgin E, 
            Song C, Jin B, Hublitz P, Eggeling C, Kohl A, Rehwinkel J
  TITLE     Infection with a Brazilian isolate of Zika virus generates RIG-I 
            stimulatory RNA and the viral NS5 protein blocks type I IFN 
            induction and signaling.
  JOURNAL   Eur J Immunol. 2018 Mar 23. doi: 10.1002/eji.201847483.
  PUBMED    29572905
REFERENCE   2  (bases 1 to 6042)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Eur J 
            Immunol. 2018 Mar 23. doi: 10.1002/eji.201847483."
COMMENT     Sequence Label: pcDNA3.2/pr-Membrane
FEATURES             Location/Qualifiers
     source          1..6042
                     /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"
     protein_bind    911..935
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1482..1506)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1532..1573
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1613..1632)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1657..1705
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1907..2335
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1994..2013)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2204..2225
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2344..2364)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2349..2678
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2529..2664
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2591..2610
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2745..3539
                     /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(2799..2818)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3409..3428
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3715..3836
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3752..3771)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3806..3825
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3885..3901)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3885..3901)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3898..3920)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3909..3925
                     /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(3933..3939,3940..3957,3958..3963))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3978..3999
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4116..4133)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4287..4875)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4367..4386)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5046..5837,5838..5906))
                     /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     5669..5688
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5907..6011)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gctggctagt
      901 taagctatca acaagtttgt acaaaaaagc aggctccgaa ttcgccctta ggaggacagc
      961 tatggcggag gtcactagac gtgggagtgc atactatatg tacttggaca gaaacgatgc
     1021 tggggaggcc atatcttttc caaccacatt ggggatgaat aagtgttata tacagatcat
     1081 ggatcttgga cacatgtgtg atgccaccat gagctatgaa tgccctatgc tggatgaggg
     1141 ggtggaacca gatgacgtcg attgttggtg caacacgacg tcaacttggg ttgtgtacgg
     1201 aacctgccat cacaaaaaag gtgaagcacg gagatctaga agagctgtga cgctcccctc
     1261 ccattccact aggaagctgc aaacgcggtc gcaaacctgg ttggaatcaa gagaatacac
     1321 aaagcacttg attagagtcg aaaattggat attcaggaac cctggcttcg cgttagcagc
     1381 agctgccatc gcttggcttt tgggaagctc aacgagccaa aaagtcatat acttggtcat
     1441 gatactgctg attgccccgg catacagcaa gggcgaattc gacccagctt tcttgtacaa
     1501 agtggttgat ctagagggcc cgcggttcga aggtaagcct atccctaacc ctctcctcgg
     1561 tctcgattct acgcgtaccg gttagtaatg agtttaaacg ggggaggcta actgaaacac
     1621 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     1681 cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     1741 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     1801 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     1861 cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc
     1921 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     1981 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2041 ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac
     2101 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2161 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2221 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg
     2281 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     2341 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2401 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2461 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     2521 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2581 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     2641 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     2701 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     2761 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     2821 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     2881 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     2941 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3001 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3061 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3121 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3181 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3241 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     3301 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     3361 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     3421 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     3481 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     3541 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     3601 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     3661 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     3721 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3781 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3841 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     3901 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3961 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4021 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4081 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4141 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4201 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4261 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4321 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4381 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4441 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     4501 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4561 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4621 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4681 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     4741 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4801 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4861 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4921 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4981 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5041 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5101 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5161 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5221 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5281 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5341 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5401 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5461 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5521 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5581 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5641 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5701 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5761 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5821 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5881 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5941 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6001 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
