LOCUS       sequence-234555-        8416 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 8416)
  TITLE     Rehwinkel Lab plasmids
REFERENCE   2  (bases 1 to 8416)
  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/NS2A
FEATURES             Location/Qualifiers
     source          1..8416
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    510..528
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    531..549
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    590..614
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1335..1359)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1412..1453
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1477..2065
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1530..1550)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1948..1959)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        2080..2396
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2247..2382
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2444..2491
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2510..2908
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             2995..3228
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3300..3421
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3337..3356)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3391..3410
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3461..3596
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3616..3635)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3617..3635)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3645..3662)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3645..3661)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3654..3676)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3803..4258
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3890..3909)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4100..4121
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4284..4388
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4389..4457,4458..5249)
                     /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(4607..4626)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5420..6008
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5909..5928
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6162..6179
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6296..6317
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6332..6337,6338..6355,6356..6362)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6370..6386
                     /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     6375..6397
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6394..6410
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6394..6410
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6429..6449
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6431..6449
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6477..6703
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6704..6884
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6931..7056
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7549..7782
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7967..8011
                     /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    8278..8395
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gcttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc tccctatcag
      541 tgatagagat cgtcgactag tccagtgtgg tggaattctg cagatatcaa caagtttgta
      601 caaaaaagca ggctccgaat tcgcccttag gaggacagct atgggatcaa ctgatcacat
      661 ggatcacttc tcccttggag tgcttgtgat tctgctcatg gtgcaggaag ggctgaagaa
      721 gagaatgacc acaaagatca tcataagcac atcaatggca gtgctggtag ctatgatcct
      781 gggaggattt tcaatgagtg acctggctaa gcttgcaatt ttgatgggcg ccaccttcgc
      841 ggaaatgaac actggaggag atgtagctca tctggcgctg atagcggcat tcaaagtcag
      901 accagcgttg ctggtatctt tcatcttcag agctaattgg acaccccgtg aaagcatgct
      961 gctggccttg gcctcgtgtc ttttgcaaac tgcgatctcc gccttggaag gcgacctgat
     1021 ggttctcatc aatggttttg ctttggcctg gttggcaata cgagcgatgg ttgttccacg
     1081 cactgataac atcaccttgg caatcctggc tgctctgaca ccactggccc ggggcacact
     1141 gcttgtggcg tggagagcag gccttgctac ttgcgggggg tttatgctcc tctctctgaa
     1201 gggaaaaggc agtgtgaaga agaacttacc atttgtcatg gccctgggac taaccgctgt
     1261 gaggctggtc gaccccatca acgtggtggg actgctgttg ctcacaagga gtgggaagcg
     1321 gaagggcgaa ttcgacccag ctttcttgta caaagtggtt gatatccagc acagtggcgg
     1381 ccgctcgagt ctagagggcc cgcggttcga aggtaagcct atccctaacc ctctcctcgg
     1441 tctcgattct acgcgtaccg gttagtaatg atcgacaatc aacctctgga ttacaaaatt
     1501 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     1561 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     1621 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     1681 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     1741 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     1801 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     1861 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     1921 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     1981 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     2041 atctcccttt gggccgcctc cccgcctggc gatggtaccg gtgtggaaag tccccaggct
     2101 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     2161 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2221 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2281 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     2341 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     2401 tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc
     2461 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca tggccaagcc
     2521 tttgtctcaa gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc
     2581 catctctgaa gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac
     2641 tggtgtcaat gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac
     2701 tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag
     2761 gggcatcttg agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat
     2821 caaagccata gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct
     2881 gccctctggt tatgtgtggg agggctaagc acaattcgag ctcggtacct ttaagaccaa
     2941 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     3001 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     3061 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     3121 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     3181 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     3241 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
     3301 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3361 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3421 atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc
     3481 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3541 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3601 gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     3661 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     3721 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     3781 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3841 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3901 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3961 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4021 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4081 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4141 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4201 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     4261 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4321 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4381 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4441 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4501 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4561 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4621 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4681 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4741 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4801 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4861 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4921 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4981 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5041 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5101 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5161 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5221 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5281 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5341 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5401 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5461 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5521 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5581 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5641 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5701 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5761 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5821 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5881 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5941 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6001 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6061 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     6121 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     6181 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     6241 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     6301 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     6361 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     6421 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa gcttaatgta
     6481 gtcttatgca atactcttgt agtcttgcaa catggtaacg atgagttagc aacatgcctt
     6541 acaaggagag aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc
     6601 cttattagga aggcaacaga cgggtctgac atggattgga cgaaccactg aattgccgca
     6661 ttgcagagat attgtattta agtgcctagc tcgatacata aacgggtctc tctggttaga
     6721 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     6781 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     6841 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
     6901 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
     6961 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
     7021 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
     7081 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
     7141 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca
     7201 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
     7261 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag
     7321 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc
     7381 accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg
     7441 gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac
     7501 caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt
     7561 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
     7621 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
     7681 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
     7741 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
     7801 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
     7861 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
     7921 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
     7981 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
     8041 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
     8101 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
     8161 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
     8221 gagagacaga gacagatcca ttcgattagt gaacggatct cgacggtatc ggttaacttt
     8281 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     8341 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     8401 taagcttggg agttcc
//
