LOCUS       sequence-235417-        8188 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Lentiviral expression of ZIKV protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8188)
  TITLE     Rehwinkel Lab plasmids
REFERENCE   2  (bases 1 to 8188)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pLenti6.3/NS4A-2K
FEATURES             Location/Qualifiers
     source          1..8188
                     /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(1107..1131)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1184..1225
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1249..1837
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1302..1322)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1720..1731)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        1852..2168
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2019..2154
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2216..2263
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2282..2680
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             2767..3000
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3072..3193
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3109..3128)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3163..3182
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3233..3368
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3388..3407)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3389..3407)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3417..3434)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3417..3433)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3426..3448)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3575..4030
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3662..3681)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3872..3893
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4056..4160
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4161..4229,4230..5021)
                     /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(4379..4398)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5192..5780
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5681..5700
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5934..5951
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6068..6089
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6104..6109,6110..6127,6128..6134)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6142..6158
                     /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     6147..6169
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6166..6182
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6166..6182
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6201..6221
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6203..6221
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6249..6475
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6476..6656
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6703..6828
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7321..7554
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7739..7783
                     /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    8050..8167
                     /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 atgggagcgg cttttggagt
      661 gatggaagcc ctgggaacac tgccaggaca catgacagag agattccagg aagccattga
      721 caacctcgct gtgctcatgc gggcagagac tggaagcagg ccttacaaag ccgcggcggc
      781 ccaattgccg gagaccctag agaccattat gcttttgggg ttgctgggaa cagtctcgct
      841 gggaatcttt ttcgtcttga tgaggaacaa gggcataggg aagatgggct ttggaatggt
      901 gactcttggg gccagcgcat ggctcatgtg gctctcggaa attgagccag ccagaattgc
      961 atgtgtcctc attgttgtgt tcctattgct ggtggtgctc atacctgagc cagaaaagca
     1021 aagatctccc caggacaacc aaatggcaat catcatcatg gtagcagtag gtcttctggg
     1081 cttgattacc gccaagggcg aattcgaccc agctttcttg tacaaagtgg ttgatatcca
     1141 gcacagtggc ggccgctcga gtctagaggg cccgcggttc gaaggtaagc ctatccctaa
     1201 ccctctcctc ggtctcgatt ctacgcgtac cggttagtaa tgatcgacaa tcaacctctg
     1261 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     1321 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     1381 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     1441 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     1501 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     1561 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     1621 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     1681 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     1741 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     1801 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gcgatggtac cggtgtggaa
     1861 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     1921 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     1981 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2041 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2101 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2161 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
     2221 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     2281 catggccaag cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat
     2341 caacagcatc cccatctctg aagactacag cgtcgccagc gcagctctct ctagcgacgg
     2401 ccgcatcttc actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt
     2461 ggtgctgggc actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg
     2521 aaatgagaac aggggcatct tgagcccctg cggacggtgc cgacaggtgc ttctcgatct
     2581 gcatcctggg atcaaagcca tagtgaagga cagtgatgga cagccgacgg cagttgggat
     2641 tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa gcacaattcg agctcggtac
     2701 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     2761 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
     2821 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     2881 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     2941 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     3001 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     3061 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3121 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3181 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc
     3241 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3301 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3361 tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg
     3421 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     3481 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     3541 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     3601 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3661 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3721 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3781 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3841 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3901 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3961 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4021 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4081 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4141 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4201 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4261 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4321 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4381 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4441 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4501 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4561 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4621 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4681 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     4741 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4801 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4861 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4921 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4981 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5041 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5101 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5161 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5221 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5281 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5341 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5401 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5461 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5521 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5581 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5641 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5701 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5761 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5821 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5881 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     5941 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     6001 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     6061 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     6121 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     6181 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc
     6241 aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta
     6301 gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg
     6361 gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac
     6421 tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc
     6481 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     6541 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     6601 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
     6661 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
     6721 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
     6781 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
     6841 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
     6901 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
     6961 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
     7021 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
     7081 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
     7141 aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat
     7201 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     7261 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     7321 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     7381 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     7441 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     7501 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     7561 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     7621 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     7681 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     7741 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     7801 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     7861 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     7921 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
     7981 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta
     8041 tcggttaact tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     8101 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     8161 aaattttatc gataagcttg ggagttcc
//
