LOCUS       sequence-229583-        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        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(3421..3445)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3498..3539
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3563..4151
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3616..3636)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4034..4045)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4166..4482
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4333..4468
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4530..4577
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4596..4994
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             5081..5314
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5386..5507
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5423..5442)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5477..5496
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5547..5682
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5702..5721)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5703..5721)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5731..5748)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5731..5747)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5740..5762)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5889..6344
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5976..5995)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6186..6207
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6370..6474
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6475..6543,6544..7335)
                     /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(6693..6712)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7506..8094
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7995..8014
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8248..8265
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8382..8403
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8418..8423,8424..8441,8442..8448)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8456..8472
                     /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     8461..8483
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8480..8496
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8480..8496
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8515..8535
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8517..8535
                     /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 ctatgtctcc ccaggacaac caaatggcaa tcatcatcat ggtagcagta ggtcttctgg
     2641 gcttgattac cgccaatgaa ctcggatggt tggagagaac aaagagtgac ctaagccatc
     2701 taatgggaag gagagaggag ggagcaacca taggattctc aatggacatt gacctgcggc
     2761 cagcctcagc ttgggccatc tatgctgcct tgacaacttt cattacccca gccgtccaac
     2821 atgcagtgac cacttcatac aacaactact ccttaatggc gatggccacg caagctggag
     2881 tgttgtttgg tatgggcaaa gggatgccat tctacgcatg ggactttgga gtcccgctgc
     2941 taatgatagg ttgctactca caattaacac ccctgaccct aatagtggcc atcattttgc
     3001 tcgtggcgca ctacatgtac ttgatcccag ggctgcaggc agcagctgcg cgtgctgccc
     3061 agaagagaac ggcagctggc atcatgaaga accctgttgt ggatggaata gtggtgactg
     3121 acattgacac aatgacaatt gacccccaag tggagaaaaa gatgggacag gtgctactca
     3181 tagcagtagc agtctccagc gccatactgt cgcggaccgc ctgggggtgg ggggaggctg
     3241 gggccctgat cacagccgca acttccactt tgtgggaagg ctctccgaac aagtactgga
     3301 actcctctac agccacttca ctgtgtaaca tttttagggg aagttacttg gctggagctt
     3361 ctctaatcta catagtaaca agaaacgctg gcttggtcaa gagacgtaag ggcgaattcg
     3421 acccagcttt cttgtacaaa gtggttgata tccagcacag tggcggccgc tcgagtctag
     3481 aggggccgcg gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc
     3541 gtaccggtta gtaatgatcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     3601 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3661 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3721 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3781 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3841 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3901 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     3961 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     4021 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4081 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4141 cgcctccccg cctggcgatg gtaccggtgt ggaaagtccc caggctcccc agcaggcaga
     4201 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4261 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4321 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4381 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4441 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4501 atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg
     4561 gcatagtata atacgacaag gtgaggaact aaaccatggc caagcctttg tctcaagaag
     4621 aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact
     4681 acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat
     4741 atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg
     4801 cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc
     4861 cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga
     4921 aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg
     4981 tgtgggaggg ctaagcacaa ttcgagctcg gtacctttaa gaccaatgac ttacaaggca
     5041 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     5101 caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     5161 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     5221 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     5281 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
     5341 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     5401 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5461 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
     5521 tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5581 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     5641 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc
     5701 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     5761 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     5821 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     5881 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     5941 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     6001 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     6061 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     6121 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     6181 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     6241 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     6301 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     6361 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6421 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     6481 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     6541 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     6601 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6661 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     6721 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6781 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6841 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6901 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6961 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     7021 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     7081 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     7141 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     7201 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     7261 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     7321 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     7381 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     7441 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     7501 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     7561 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     7621 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     7681 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7741 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7801 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7861 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7921 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7981 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     8041 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     8101 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     8161 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     8221 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     8281 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     8341 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     8401 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     8461 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     8521 aaccctcact aaagggaaca aaagctggag ctgcaagctt
//
