LOCUS       sequence-229577-        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        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(3352..3376)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3429..3470
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3494..4082
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3547..3567)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3965..3976)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4097..4413
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4264..4399
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4461..4508
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4527..4925
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             5012..5245
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5317..5438
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5354..5373)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5408..5427
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5478..5613
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5633..5652)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5634..5652)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5662..5679)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5662..5678)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5671..5693)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5820..6275
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5907..5926)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6117..6138
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6301..6405
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6406..6474,6475..7266)
                     /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(6624..6643)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7437..8025
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7926..7945
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8179..8196
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8313..8334
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8349..8354,8355..8372,8373..8379)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8387..8403
                     /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     8392..8414
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8411..8427
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8411..8427
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8446..8466
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8448..8466
                     /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 ctatgaatga actcggatgg ttggagagaa caaagagtga cctaagccat ctaatgggaa
     2641 ggagagagga gggagcaacc ataggattct caatggacat tgacctgcgg ccagcctcag
     2701 cttgggccat ctatgctgcc ttgacaactt tcattacccc agccgtccaa catgcagtga
     2761 ccacttcata caacaactac tccttaatgg cgatggccac gcaagctgga gtgttgtttg
     2821 gtatgggcaa agggatgcca ttctacgcat gggactttgg agtcccgctg ctaatgatag
     2881 gttgctactc acaattaaca cccctgaccc taatagtggc catcattttg ctcgtggcgc
     2941 actacatgta cttgatccca gggctgcagg cagcagctgc gcgtgctgcc cagaagagaa
     3001 cggcagctgg catcatgaag aaccctgttg tggatggaat agtggtgact gacattgaca
     3061 caatgacaat tgacccccaa gtggagaaaa agatgggaca ggtgctactc atagcagtag
     3121 cagtctccag cgccatactg tcgcggaccg cctgggggtg gggggaggct ggggccctga
     3181 tcacagccgc aacttccact ttgtgggaag gctctccgaa caagtactgg aactcctcta
     3241 cagccacttc actgtgtaac atttttaggg gaagttactt ggctggagct tctctaatct
     3301 acatagtaac aagaaacgct ggcttggtca agagacgtaa gggcgaattc gacccagctt
     3361 tcttgtacaa agtggttgat atccagcaca gtggcggccg ctcgagtcta gaggggccgc
     3421 ggttcgaagg taagcctatc cctaaccctc tcctcggtct cgattctacg cgtaccggtt
     3481 agtaatgatc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3541 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3601 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3661 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3721 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     3781 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3841 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     3901 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3961 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     4021 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     4081 gcctggcgat ggtaccggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4141 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4201 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4261 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4321 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4381 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4441 ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat
     4501 aatacgacaa ggtgaggaac taaaccatgg ccaagccttt gtctcaagaa gaatccaccc
     4561 tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg
     4621 ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta
     4681 ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca
     4741 acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac
     4801 ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg
     4861 atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg
     4921 gctaagcaca attcgagctc ggtaccttta agaccaatga cttacaaggc agctgtagat
     4981 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     5041 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     5101 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     5161 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     5221 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     5281 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     5341 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5401 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     5461 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     5521 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     5581 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag
     5641 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     5701 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     5761 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     5821 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     5881 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5941 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     6001 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     6061 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     6121 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     6181 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     6241 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     6301 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     6361 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     6421 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6481 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     6541 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     6601 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     6661 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6721 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6781 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6841 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6901 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6961 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     7021 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     7081 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     7141 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     7201 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     7261 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     7321 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     7381 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7441 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7501 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7561 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     7621 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7681 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7741 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7801 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7861 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7921 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7981 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     8041 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     8101 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     8161 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     8221 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     8281 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     8341 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     8401 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
     8461 taaagggaac aaaagctgga gctgcaagct t
//
