LOCUS       sequence-229572-        8242 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 8242)
  TITLE     Rehwinkel Lab plasmids
REFERENCE   2  (bases 1 to 8242)
  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/pr-Membrane
FEATURES             Location/Qualifiers
     source          1..8242
                     /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(3103..3127)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3180..3221
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3245..3833
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3298..3318)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3716..3727)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        3848..4164
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4015..4150
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4212..4259
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4278..4676
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4763..4996
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5068..5189
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5105..5124)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5159..5178
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5229..5364
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5384..5403)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5385..5403)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5413..5430)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5413..5429)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5422..5444)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5571..6026
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5658..5677)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5868..5889
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6052..6156
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6157..6225,6226..7017)
                     /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(6375..6394)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7188..7776
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7677..7696
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7930..7947
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8064..8085
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8100..8105,8106..8123,8124..8130)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8138..8154
                     /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     8143..8165
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8162..8178
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8162..8178
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8197..8217
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8199..8217
                     /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 ctatggcgga ggtcactaga cgtgggagtg catactatat gtacttggac agaaacgatg
     2641 ctggggaggc catatctttt ccaaccacat tggggatgaa taagtgttat atacagatca
     2701 tggatcttgg acacatgtgt gatgccacca tgagctatga atgccctatg ctggatgagg
     2761 gggtggaacc agatgacgtc gattgttggt gcaacacgac gtcaacttgg gttgtgtacg
     2821 gaacctgcca tcacaaaaaa ggtgaagcac ggagatctag aagagctgtg acgctcccct
     2881 cccattccac taggaagctg caaacgcggt cgcaaacctg gttggaatca agagaataca
     2941 caaagcactt gattagagtc gaaaattgga tattcaggaa ccctggcttc gcgttagcag
     3001 cagctgccat cgcttggctt ttgggaagct caacgagcca aaaagtcata tacttggtca
     3061 tgatactgct gattgccccg gcatacagca agggcgaatt cgacccagct ttcttgtaca
     3121 aagtggttga tatccagcac agtggcggcc gctcgagtct agaggggccg cggttcgaag
     3181 gtaagcctat ccctaaccct ctcctcggtc tcgattctac gcgtaccggt tagtaatgat
     3241 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3301 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3361 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     3421 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     3481 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     3541 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     3601 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     3661 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     3721 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     3781 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggcga
     3841 tggtaccggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3901 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3961 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4021 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4081 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4141 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4201 gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     4261 aggtgaggaa ctaaaccatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa
     4321 gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag
     4381 ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac
     4441 cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt
     4501 gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac
     4561 aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc
     4621 cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaagcac
     4681 aattcgagct cggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     4741 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
     4801 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     4861 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     4921 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     4981 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     5041 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     5101 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5161 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     5221 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5281 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     5341 agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta
     5401 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     5461 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     5521 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     5581 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     5641 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5701 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5761 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5821 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5881 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5941 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     6001 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     6061 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     6121 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     6181 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6241 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6301 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6361 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6421 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6481 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6541 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6601 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6661 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6721 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6781 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6841 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6901 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6961 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     7021 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     7081 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     7141 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     7201 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7261 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7321 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7381 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7441 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7501 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7561 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7621 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7681 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7741 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7801 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7861 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     7921 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     7981 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     8041 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     8101 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     8161 acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa
     8221 caaaagctgg agctgcaagc tt
//
