LOCUS       sequence-229582-        8119 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of ZIKV protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8119)
  TITLE     Rehwinkel Lab plasmids
REFERENCE   2  (bases 1 to 8119)
  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"
COMMENT     Sequence Label: pLenti6.3/NS4A
FEATURES             Location/Qualifiers
     source          1..8119
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1075..1308
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1804..1921
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2247..2450
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2401..2421
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2452..2470
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2473..2491
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2532..2556
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2980..3004)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3057..3098
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3122..3710
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3175..3195)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3593..3604)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        3725..4041
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3892..4027
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4089..4136
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4155..4553
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4640..4873
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4945..5066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4982..5001)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5036..5055
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5106..5241
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5261..5280)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5262..5280)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5290..5307)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5290..5306)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5299..5321)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5448..5903
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5535..5554)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5745..5766
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5929..6033
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6034..6102,6103..6894)
                     /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     complement(6252..6271)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7065..7653
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7554..7573
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7807..7824
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7941..7962
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7977..7982,7983..8000,8001..8007)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8015..8031
                     /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)."
     primer_bind     8020..8042
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8039..8055
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8039..8055
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8074..8094
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8076..8094
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgataag cttgggagtt ccgcttacat aacttacggt aaatggcccg cctggctgac
     1981 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     2041 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     2101 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     2161 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     2221 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     2281 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     2341 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     2401 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctccctatca
     2461 gtgatagaga tctccctatc agtgatagag atcgtcgact agtccagtgt ggtggaattc
     2521 tgcagatatc aacaagtttg tacaaaaaag caggctccga attcgccctt aggaggacag
     2581 ctatgggagc ggcttttgga gtgatggaag ccctgggaac actgccagga cacatgacag
     2641 agagattcca ggaagccatt gacaacctcg ctgtgctcat gcgggcagag actggaagca
     2701 ggccttacaa agccgcggcg gcccaattgc cggagaccct agagaccatt atgcttttgg
     2761 ggttgctggg aacagtctcg ctgggaatct ttttcgtctt gatgaggaac aagggcatag
     2821 ggaagatggg ctttggaatg gtgactcttg gggccagcgc atggctcatg tggctctcgg
     2881 aaattgagcc agccagaatt gcatgtgtcc tcattgttgt gttcctattg ctggtggtgc
     2941 tcatacctga gccagaaaag caaagaaagg gcgaattcga cccagctttc ttgtacaaag
     3001 tggttgatat ccagcacagt ggcggccgct cgagtctaga ggggccgcgg ttcgaaggta
     3061 agcctatccc taaccctctc ctcggtctcg attctacgcg taccggttag taatgatcga
     3121 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     3181 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     3241 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     3301 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     3361 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     3421 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     3481 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     3541 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     3601 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3661 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggcgatgg
     3721 taccggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3781 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3841 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3901 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3961 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4021 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4081 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     4141 tgaggaacta aaccatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag
     4201 caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc
     4261 tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt
     4321 gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta
     4381 tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg
     4441 tgcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga
     4501 cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taagcacaat
     4561 tcgagctcgg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     4621 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt
     4681 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     4741 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     4801 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     4861 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     4921 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
     4981 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5041 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
     5101 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5161 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     5221 gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta
     5281 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     5341 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     5401 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     5461 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     5521 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     5581 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     5641 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     5701 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5761 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     5821 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     5881 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     5941 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     6001 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     6061 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     6121 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     6181 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     6241 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     6301 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     6361 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     6421 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     6481 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     6541 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     6601 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     6661 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6721 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6781 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6841 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6901 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6961 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     7021 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     7081 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     7141 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     7201 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7261 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     7321 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     7381 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     7441 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     7501 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7561 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     7621 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     7681 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     7741 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     7801 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     7861 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     7921 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     7981 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     8041 ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa
     8101 aagctggagc tgcaagctt
//
