LOCUS       sequence-241012-        8491 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 8491)
  TITLE     Rehwinkel Lab plasmids
REFERENCE   2  (bases 1 to 8491)
  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/NS4B
FEATURES             Location/Qualifiers
     source          1..8491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    206..224
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    227..245
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    286..310
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1106..1130)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1183..1224
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1248..1836
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1301..1321)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1719..1730)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        1851..2167
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2018..2153
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2215..2262
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2281..2679
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             2766..2999
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3071..3192
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3108..3127)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3162..3181
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3232..3367
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3387..3406)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3388..3406)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3416..3433)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3416..3432)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3425..3447)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3574..4029
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3661..3680)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3871..3892
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4055..4159
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4160..4228,4229..5020)
                     /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(4378..4397)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5191..5779
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5680..5699
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5933..5950
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6067..6088
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6103..6108,6109..6126,6127..6133)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6141..6157
                     /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     6146..6168
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6165..6181
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6165..6181
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6200..6220
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6202..6220
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6248..6474
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6475..6655
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6702..6827
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7320..7553
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7738..7782
                     /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    8049..8166
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctctccct atcagtgata gagatctccc tatcagtgat
      241 agagatcgtc gactagtcca gtgtggtgga attctgcaga tatcaacaag tttgtacaaa
      301 aaagcaggct ccgaattcgc ccttaggagg acagctatga atgaactcgg atggttggag
      361 agaacaaaga gtgacctaag ccatctaatg ggaaggagag aggagggagc aaccatagga
      421 ttctcaatgg acattgacct gcggccagcc tcagcttggg ccatctatgc tgccttgaca
      481 actttcatta ccccagccgt ccaacatgca gtgaccactt catacaacaa ctactcctta
      541 atggcgatgg ccacgcaagc tggagtgttg tttggtatgg gcaaagggat gccattctac
      601 gcatgggact ttggagtccc gctgctaatg ataggttgct actcacaatt aacacccctg
      661 accctaatag tggccatcat tttgctcgtg gcgcactaca tgtacttgat cccagggctg
      721 caggcagcag ctgcgcgtgc tgcccagaag agaacggcag ctggcatcat gaagaaccct
      781 gttgtggatg gaatagtggt gactgacatt gacacaatga caattgaccc ccaagtggag
      841 aaaaagatgg gacaggtgct actcatagca gtagcagtct ccagcgccat actgtcgcgg
      901 accgcctggg ggtgggggga ggctggggcc ctgatcacag ccgcaacttc cactttgtgg
      961 gaaggctctc cgaacaagta ctggaactcc tctacagcca cttcactgtg taacattttt
     1021 aggggaagtt acttggctgg agcttctcta atctacatag taacaagaaa cgctggcttg
     1081 gtcaagagac gtaagggcga attcgaccca gctttcttgt acaaagtggt tgatatccag
     1141 cacagtggcg gccgctcgag tctagagggc ccgcggttcg aaggtaagcc tatccctaac
     1201 cctctcctcg gtctcgattc tacgcgtacc ggttagtaat gatcgacaat caacctctgg
     1261 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     1321 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     1381 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     1441 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     1501 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     1561 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     1621 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     1681 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     1741 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     1801 agacgagtcg gatctccctt tgggccgcct ccccgcctgg cgatggtacc ggtgtggaaa
     1861 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     1921 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     1981 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     2041 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     2101 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     2161 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
     2221 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     2281 atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc
     2341 aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc
     2401 cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg
     2461 gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga
     2521 aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg
     2581 catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt
     2641 cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag cacaattcga gctcggtacc
     2701 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     2761 gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg
     2821 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     2881 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     2941 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     3001 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     3061 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3121 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3181 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc
     3241 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3301 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3361 ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
     3421 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     3481 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     3541 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     3601 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3661 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3721 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3781 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3841 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3901 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3961 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     4021 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4081 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4141 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4201 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4261 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4321 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4381 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4441 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4501 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4561 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4621 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4681 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4741 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4801 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4861 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4921 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4981 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5041 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5101 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5161 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5221 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5281 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5341 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5401 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5461 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5521 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5581 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5641 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5701 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5761 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5821 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5881 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     5941 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     6001 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     6061 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     6121 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     6181 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca
     6241 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
     6301 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
     6361 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
     6421 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
     6481 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     6541 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     6601 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     6661 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     6721 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     6781 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     6841 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     6901 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     6961 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     7021 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     7081 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     7141 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     7201 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     7261 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     7321 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     7381 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     7441 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     7501 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     7561 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     7621 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     7681 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     7741 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     7801 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     7861 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     7921 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     7981 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat
     8041 cggttaactt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     8101 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     8161 aattttatcg ataagcttgg gagttccgct tacataactt acggtaaatg gcccgcctgg
     8221 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     8281 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     8341 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     8401 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     8461 catctacgta ttagtcatcg ctattaccat g
//
