LOCUS       sequence-358511-        5993 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 5993)
  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 5993)
  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-PrM
FEATURES             Location/Qualifiers
     source          1..5993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(81..101)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        86..415
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      266..401
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     328..347
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             482..1276
                     /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(536..555)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1146..1165
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1450..1571
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1487..1506)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1541..1560
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1620..1636)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1620..1636)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1633..1655)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1644..1660
                     /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(1668..1674,1675..1692,1693..1698))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1713..1734
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1851..1868)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2022..2610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2102..2121)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2781..3572,3573..3641))
                     /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     3404..3423
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3642..3746)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3821..3840)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4012..4391
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4392..4595
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4546..4566
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4640..4659
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4640..4658
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4715..4724
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5297..5320
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5361..5378)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5367..5591
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(5637..5993,1..72)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5724..5743)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5934..5955
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
       61 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
      121 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
      181 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
      241 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
      301 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
      361 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
      421 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
      481 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
      541 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
      601 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
      661 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
      721 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
      781 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
      841 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
      901 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
      961 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     1021 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     1081 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     1141 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     1201 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     1261 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     1321 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     1381 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     1441 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     1501 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     1561 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     1621 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     1681 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     1741 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     1801 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     1861 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1921 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1981 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2041 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2101 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2161 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2221 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2281 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2341 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2401 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2461 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2521 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2581 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2641 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2701 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2761 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2821 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2881 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     2941 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3001 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3061 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3121 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3181 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3241 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3301 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3361 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3421 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3481 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3541 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3601 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3661 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3721 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     3781 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     3841 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     3901 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     3961 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
     4021 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     4081 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     4141 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     4201 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     4261 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     4321 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     4381 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     4441 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     4501 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     4561 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg
     4621 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct ggctagcgtt
     4681 taaacttaag cttggtaccg agctcggatc cgccgccgcc atggagaaga agagacgagg
     4741 cgcagatact agtgtcggaa ttgttggcct cctgctgacc acagctatgg cagcggaggt
     4801 cactagacgt gggagtgcat actatatgta cttggacaga aacgatgctg gggaggccat
     4861 atcttttcca accacattgg ggatgaataa gtgttatata cagatcatgg atcttggaca
     4921 catgtgtgat gccaccatga gctatgaatg ccctatgctg gatgaggggg tggaaccaga
     4981 tgacgtcgat tgttggtgca acacgacgtc aacttgggtt gtgtacggaa cctgccatca
     5041 caaaaaaggt gaagcacgga gatctagaag agctgtgacg ctcccctccc attccaccag
     5101 gaagctgcaa acgcggtcgc aaacctggtt ggaatcaaga gaatacacaa agcacttgat
     5161 tagagtcgaa aattggatat tcaggaaccc tggcttcgcg ttagcagcag ctgccatcgc
     5221 ttggcttttg ggaagctcaa cgagccaaaa agtcatatac ttggtcatga tactgctgat
     5281 tgccccggca tacagcgact acaaagacga tgacgacaag tagctcgagt ctagagggcc
     5341 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     5401 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     5461 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     5521 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     5581 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     5641 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5701 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5761 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5821 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     5881 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     5941 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg att
//
