LOCUS       sequence-358512-        6098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of ZIKV ORF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6098)
  AUTHORS   Sen Santara S, Crespo AC, Mulik S, Ovies C, Boulenouar S, Strominger
            JL, Lieberman J
  TITLE     Decidual NK cells kill Zika virus-infected trophoblasts.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Nov 23;118(47):e2115410118. doi: 
            10.1073/pnas.2115410118.
  PUBMED    34785597
REFERENCE   2  (bases 1 to 6098)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2021-11-23"; volume: "118(47):e2115410118. 
            doi"; pages: " 10.1073/pnas.2115410118"
COMMENT     Sequence Label: pFLAG-CMV4-ZIKV-NS2A
FEATURES             Location/Qualifiers
     source          1..6098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        243..446
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     397..417
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     491..510
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        491..509
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      566..575
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1253..1276
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1317..1334)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1323..1547
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1593..2021
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1680..1699)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1890..1911
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2030..2050)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2035..2364
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2215..2350
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2277..2296
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2431..3225
                     /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(2485..2504)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3095..3114
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3399..3520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3436..3455)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3490..3509
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3569..3585)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3569..3585)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3582..3604)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3593..3609
                     /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(3617..3623,3624..3641,3642..3647))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3662..3683
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3800..3817)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3971..4559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4051..4070)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4730..5521,5522..5590))
                     /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     5353..5372
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5591..5695)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5770..5789)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(5961..6098,1..242)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
       61 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      121 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      181 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      241 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      301 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      361 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      421 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc
      481 ttatcgaaat taatacgact cactataggg agacccaagc tggctagcgt ttaaacttaa
      541 gcttggtacc gagctcggat ccgccgccgc catgggatca actgatcaca tggaccactt
      601 ctcccttgga gtgcttgtga tcctgctcat ggtgcaggaa gggctgaaga agagaatgac
      661 cacaaagatc atcataagca catcaatggc agtgctggta gctatgatcc tgggaggatt
      721 ttcaatgagt gacctggcta agcttgcaat tttgatgggt gccaccttcg cggaaatgaa
      781 cactggagga gatgtagctc atctggcgct gatagcggca ttcaaagtca gaccagcgtt
      841 gctggtatct ttcatcttca gagctaattg gacaccccgt gaaagcatgc tgctggcctt
      901 ggcctcgtgt cttttgcaaa ctgcgatctc cgccttggaa ggcgacctga tggttctcat
      961 caatggtttt gctttggcct ggttggcaat acgagcgatg gttgttccac gcactgataa
     1021 catcaccttg gcaatcctgg ctgctctgac accactggcc cggggcacac tgcttgtggc
     1081 gtggagagca ggccttgcta cttgcggggg gtttatgctc ctctctctga agggaaaagg
     1141 cagtgtgaag aagaacttac catttgtcat ggccctggga ctaaccgctg tgaggctggt
     1201 cgaccccatc aacgtggtgg gactgctgtt gctcacaagg agtgggaagc gggactacaa
     1261 agacgatgac gacaagtagc tcgagtctag agggcccgtt taaacccgct gatcagcctc
     1321 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     1381 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     1441 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     1501 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     1561 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     1621 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     1681 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     1741 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     1801 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     1861 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     1921 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     1981 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     2041 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2101 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2161 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     2221 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     2281 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     2341 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     2401 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     2461 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     2521 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     2581 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     2641 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     2701 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     2761 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     2821 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     2881 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     2941 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3001 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3061 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3121 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3181 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     3241 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     3301 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     3361 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     3421 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3481 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     3541 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     3601 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3661 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3721 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3781 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3841 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3901 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3961 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4021 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4081 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4141 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4201 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4261 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4321 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4381 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4441 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4501 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4561 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4621 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4681 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4741 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4801 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4861 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4921 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4981 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5041 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5101 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5161 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5221 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5281 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5341 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5401 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5461 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5521 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5581 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5641 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5701 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg
     5761 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     5821 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     5881 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
     5941 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
     6001 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     6061 cccgcctggc tgaccgccca acgacccccg cccattga
//
