LOCUS       sequence-234597-        8119 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 8119)
  TITLE     Rehwinkel Lab plasmids
REFERENCE   2  (bases 1 to 8119)
  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/NS4A
FEATURES             Location/Qualifiers
     source          1..8119
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    846..1079
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1264..1308
                     /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    1575..1692
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2018..2221
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2172..2192
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2223..2241
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2244..2262
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2303..2327
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2751..2775)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2828..2869
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    2893..3481
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2946..2966)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3364..3375)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        3496..3812
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3663..3798
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3860..3907
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3926..4324
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4411..4644
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4716..4837
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4753..4772)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4807..4826
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4877..5012
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5032..5051)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5033..5051)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5061..5078)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5061..5077)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5070..5092)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5219..5674
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5306..5325)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5516..5537
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5700..5804
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5805..5873,5874..6665)
                     /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(6023..6042)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6836..7424
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7325..7344
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7578..7595
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7712..7733
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7748..7753,7754..7771,7772..7778)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7786..7802
                     /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     7791..7813
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7810..7826
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7810..7826
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7845..7865
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7847..7865
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7893..8119
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1441 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     1501 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga
     1561 cggtatcggt taacttttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     1621 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     1681 attcaaaatt ttatcgataa gcttgggagt tccgcttaca taacttacgg taaatggccc
     1741 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     1801 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     1861 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     1921 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     1981 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     2041 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     2101 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     2161 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     2221 tctccctatc agtgatagag atctccctat cagtgataga gatcgtcgac tagtccagtg
     2281 tggtggaatt ctgcagatat caacaagttt gtacaaaaaa gcaggctccg aattcgccct
     2341 taggaggaca gctatgggag cggcttttgg agtgatggaa gccctgggaa cactgccagg
     2401 acacatgaca gagagattcc aggaagccat tgacaacctc gctgtgctca tgcgggcaga
     2461 gactggaagc aggccttaca aagccgcggc ggcccaattg ccggagaccc tagagaccat
     2521 tatgcttttg gggttgctgg gaacagtctc gctgggaatc tttttcgtct tgatgaggaa
     2581 caagggcata gggaagatgg gctttggaat ggtgactctt ggggccagcg catggctcat
     2641 gtggctctcg gaaattgagc cagccagaat tgcatgtgtc ctcattgttg tgttcctatt
     2701 gctggtggtg ctcatacctg agccagaaaa gcaaagaaag ggcgaattcg acccagcttt
     2761 cttgtacaaa gtggttgata tccagcacag tggcggccgc tcgagtctag agggcccgcg
     2821 gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtta
     2881 gtaatgatcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2941 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     3001 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     3061 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     3121 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     3181 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     3241 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     3301 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     3361 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     3421 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     3481 cctggcgatg gtaccggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3541 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3601 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3661 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3721 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3781 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3841 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
     3901 atacgacaag gtgaggaact aaaccatggc caagcctttg tctcaagaag aatccaccct
     3961 cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact acagcgtcgc
     4021 cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac
     4081 tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa
     4141 cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg
     4201 gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga
     4261 tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg
     4321 ctaagcacaa ttcgagctcg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     4381 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
     4441 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     4501 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     4561 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     4621 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     4681 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     4741 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4801 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     4861 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4921 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4981 ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt
     5041 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     5101 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     5161 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     5221 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5281 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5341 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     5401 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5461 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5521 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5581 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5641 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     5701 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5761 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     5821 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5881 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5941 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     6001 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     6061 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     6121 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     6181 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     6241 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     6301 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     6361 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     6421 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     6481 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6541 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6601 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6661 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     6721 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6781 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6841 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6901 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6961 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     7021 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     7081 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     7141 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     7201 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     7261 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     7321 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     7381 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     7441 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     7501 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     7561 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     7621 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     7681 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     7741 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     7801 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
     7861 aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc
     7921 ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc
     7981 atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg
     8041 tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg
     8101 cctagctcga tacataaac
//
