LOCUS       sequence-234606-        8128 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 8128)
  TITLE     Rehwinkel Lab plasmids
REFERENCE   2  (bases 1 to 8128)
  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/NS2B
FEATURES             Location/Qualifiers
     source          1..8128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1491..1535
                     /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    1802..1919
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2245..2448
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2399..2419
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2450..2468
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2471..2489
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2530..2554
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2987..3011)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3064..3105
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3129..3717
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3182..3202)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3600..3611)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        3732..4048
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3899..4034
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4096..4143
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4162..4560
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4647..4880
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4952..5073
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4989..5008)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5043..5062
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5113..5248
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5268..5287)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5269..5287)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5297..5314)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5297..5313)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5306..5328)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5455..5910
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5542..5561)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5752..5773
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5936..6040
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6041..6109,6110..6901)
                     /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(6259..6278)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7072..7660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7561..7580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7814..7831
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7948..7969
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7984..7989,7990..8007,8008..8014)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8022..8038
                     /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     8027..8049
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8046..8062
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8046..8062
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8081..8101
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8083..8101
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     1741 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcggttaa
     1801 cttttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     1861 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaatttta
     1921 tcgataagct tgggagttcc gcttacataa cttacggtaa atggcccgcc tggctgaccg
     1981 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     2041 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     2101 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     2161 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     2221 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2281 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2341 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2401 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt
     2461 gatagagatc tccctatcag tgatagagat cgtcgactag tccagtgtgg tggaattctg
     2521 cagatatcaa caagtttgta caaaaaagca ggctccgaat tcgcccttag gaggacagct
     2581 atgagctggc cccctagcga agtactcaca gctgttggcc tgatatgcgc attggctgga
     2641 gggttcgcca aggcagatat agagatggct gggcccatgg ccgcggtcgg tctgctaatt
     2701 gtcagttacg tggtctcagg aaagagtgtg gacatgtaca ttgaaagagc aggtgacatc
     2761 acatgggaaa aagatgcgga agtcactgga aacagtcccc ggctcgatgt ggcgctagat
     2821 gagagtggtg atttctccct ggtggaggat gacggtcccc ccatgagaga gatcatactc
     2881 aaggtggtcc tgatgaccat ctgtggcatg aacccaatag ccataccctt tgcagctgga
     2941 gcgtggtacg tatacgtgaa gactggaaaa aggaagggcg aattcgaccc agctttcttg
     3001 tacaaagtgg ttgatatcca gcacagtggc ggccgctcga gtctagaggg cccgcggttc
     3061 gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggttagtaa
     3121 tgatcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     3181 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     3241 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     3301 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     3361 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     3421 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     3481 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat
     3541 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3601 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     3661 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     3721 gcgatggtac cggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3781 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3841 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3901 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3961 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     4021 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     4081 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac
     4141 gacaaggtga ggaactaaac catggccaag cctttgtctc aagaagaatc caccctcatt
     4201 gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag cgtcgccagc
     4261 gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca ttttactggg
     4321 ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc tggcaacctg
     4381 acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg cggacggtgc
     4441 cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga cagtgatgga
     4501 cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa
     4561 gcacaattcg agctcggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     4621 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     4681 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     4741 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     4801 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     4861 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     4921 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     4981 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5041 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     5101 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5161 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     5221 aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt
     5281 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     5341 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     5401 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     5461 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5521 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5581 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5641 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     5701 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5761 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     5821 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5881 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     5941 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     6001 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     6061 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     6121 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     6181 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     6241 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     6301 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     6361 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6421 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     6481 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6541 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6601 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6661 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6721 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6781 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6841 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6901 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6961 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     7021 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     7081 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     7141 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     7201 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     7261 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     7321 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     7381 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7441 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     7501 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7561 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7621 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7681 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     7741 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     7801 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     7861 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     7921 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     7981 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     8041 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag
     8101 ggaacaaaag ctggagctgc aagcttaa
//
