LOCUS       sequence-234643-        8560 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 8560)
  TITLE     Rehwinkel Lab plasmids
REFERENCE   2  (bases 1 to 8560)
  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: pLenti6.3/2K-NS4B
FEATURES             Location/Qualifiers
     source          1..8560
                     /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(1175..1199)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1252..1293
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1317..1905
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1370..1390)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1788..1799)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        1920..2236
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2087..2222
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2284..2331
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2350..2748
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             2835..3068
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3140..3261
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3177..3196)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3231..3250
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3301..3436
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3456..3475)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3457..3475)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3485..3502)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3485..3501)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3494..3516)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3643..4098
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3730..3749)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3940..3961
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4124..4228
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4229..4297,4298..5089)
                     /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(4447..4466)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5260..5848
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5749..5768
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6002..6019
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6136..6157
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6172..6177,6178..6195,6196..6202)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6210..6226
                     /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     6215..6237
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6234..6250
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6234..6250
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6269..6289
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6271..6289
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6317..6543
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6544..6724
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6771..6896
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7389..7622
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7807..7851
                     /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    8118..8235
                     /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 acagctatgt ctccccagga caaccaaatg
      361 gcaatcatca tcatggtagc agtaggtctt ctgggcttga ttaccgccaa tgaactcgga
      421 tggttggaga gaacaaagag tgacctaagc catctaatgg gaaggagaga ggagggagca
      481 accataggat tctcaatgga cattgacctg cggccagcct cagcttgggc catctatgct
      541 gccttgacaa ctttcattac cccagccgtc caacatgcag tgaccacttc atacaacaac
      601 tactccttaa tggcgatggc cacgcaagct ggagtgttgt ttggtatggg caaagggatg
      661 ccattctacg catgggactt tggagtcccg ctgctaatga taggttgcta ctcacaatta
      721 acacccctga ccctaatagt ggccatcatt ttgctcgtgg cgcactacat gtacttgatc
      781 ccagggctgc aggcagcagc tgcgcgtgct gcccagaaga gaacggcagc tggcatcatg
      841 aagaaccctg ttgtggatgg aatagtggtg actgacattg acacaatgac aattgacccc
      901 caagtggaga aaaagatggg acaggtgcta ctcatagcag tagcagtctc cagcgccata
      961 ctgtcgcgga ccgcctgggg gtggggggag gctggggccc tgatcacagc cgcaacttcc
     1021 actttgtggg aaggctctcc gaacaagtac tggaactcct ctacagccac ttcactgtgt
     1081 aacattttta ggggaagtta cttggctgga gcttctctaa tctacatagt aacaagaaac
     1141 gctggcttgg tcaagagacg taagggcgaa ttcgacccag ctttcttgta caaagtggtt
     1201 gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgcggttcga aggtaagcct
     1261 atccctaacc ctctcctcgg tctcgattct acgcgtaccg gttagtaatg atcgacaatc
     1321 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     1381 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     1441 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     1501 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     1561 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     1621 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     1681 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     1741 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     1801 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     1861 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctggc gatggtaccg
     1921 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     1981 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2041 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     2101 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     2161 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     2221 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca
     2281 cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg
     2341 aactaaacca tggccaagcc tttgtctcaa gaagaatcca ccctcattga aagagcaacg
     2401 gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc agctctctct
     2461 agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg accttgtgca
     2521 gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc
     2581 gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg acaggtgctt
     2641 ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca gccgacggca
     2701 gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaagc acaattcgag
     2761 ctcggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     2821 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc
     2881 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     2941 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     3001 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     3061 ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     3121 aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa
     3181 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3241 caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc
     3301 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3361 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3421 ggcttttttg gaggcctagg gacgtaccca attcgcccta tagtgagtcg tattacgcgc
     3481 gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     3541 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     3601 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     3661 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3721 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3781 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3841 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3901 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3961 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4021 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4081 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4141 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4201 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4261 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4321 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4381 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4441 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4501 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4561 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4621 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4681 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4741 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4801 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4861 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4921 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4981 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5041 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5101 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5161 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5221 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5281 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5341 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5401 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5461 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5521 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5581 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5641 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5701 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5761 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5821 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5881 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5941 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6001 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     6061 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     6121 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     6181 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     6241 cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct
     6301 ggagctgcaa gcttaatgta gtcttatgca atactcttgt agtcttgcaa catggtaacg
     6361 atgagttagc aacatgcctt acaaggagag aaaaagcacc gtgcatgccg attggtggaa
     6421 gtaaggtggt acgatcgtgc cttattagga aggcaacaga cgggtctgac atggattgga
     6481 cgaaccactg aattgccgca ttgcagagat attgtattta agtgcctagc tcgatacata
     6541 aacgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     6601 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     6661 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     6721 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     6781 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     6841 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     6901 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     6961 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     7021 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     7081 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     7141 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     7201 agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga
     7261 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     7321 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     7381 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     7441 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     7501 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     7561 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     7621 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     7681 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     7741 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     7801 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     7861 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     7921 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     7981 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     8041 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatct
     8101 cgacggtatc ggttaacttt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     8161 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     8221 aaaattcaaa attttatcga taagcttggg agttccgctt acataactta cggtaaatgg
     8281 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     8341 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     8401 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     8461 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     8521 ttggcagtac atctacgtat tagtcatcgc tattaccatg
//
