LOCUS       sequence-229575-        8188 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 8188)
  TITLE     Rehwinkel Lab plasmids
REFERENCE   2  (bases 1 to 8188)
  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-2K
FEATURES             Location/Qualifiers
     source          1..8188
                     /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(3049..3073)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3126..3167
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3191..3779
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3244..3264)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3662..3673)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        3794..4110
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3961..4096
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4158..4205
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4224..4622
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4709..4942
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5014..5135
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5051..5070)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5105..5124
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5175..5310
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5330..5349)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5331..5349)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5359..5376)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5359..5375)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5368..5390)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5517..5972
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5604..5623)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5814..5835
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5998..6102
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6103..6171,6172..6963)
                     /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(6321..6340)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7134..7722
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7623..7642
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7876..7893
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8010..8031
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8046..8051,8052..8069,8070..8076)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8084..8100
                     /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     8089..8111
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8108..8124
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8108..8124
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8143..8163
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8145..8163
                     /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 ctatgggagc ggcttttgga gtgatggaag ccctgggaac actgccagga cacatgacag
     2641 agagattcca ggaagccatt gacaacctcg ctgtgctcat gcgggcagag actggaagca
     2701 ggccttacaa agccgcggcg gcccaattgc cggagaccct agagaccatt atgcttttgg
     2761 ggttgctggg aacagtctcg ctgggaatct ttttcgtctt gatgaggaac aagggcatag
     2821 ggaagatggg ctttggaatg gtgactcttg gggccagcgc atggctcatg tggctctcgg
     2881 aaattgagcc agccagaatt gcatgtgtcc tcattgttgt gttcctattg ctggtggtgc
     2941 tcatacctga gccagaaaag caaagatctc cccaggacaa ccaaatggca atcatcatca
     3001 tggtagcagt aggtcttctg ggcttgatta ccgccaaggg cgaattcgac ccagctttct
     3061 tgtacaaagt ggttgatatc cagcacagtg gcggccgctc gagtctagag gggccgcggt
     3121 tcgaaggtaa gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggttagt
     3181 aatgatcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     3241 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     3301 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     3361 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     3421 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     3481 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     3541 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     3601 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3661 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3721 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     3781 tggcgatggt accggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3841 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3901 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3961 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4021 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4081 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4141 ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat
     4201 acgacaaggt gaggaactaa accatggcca agcctttgtc tcaagaagaa tccaccctca
     4261 ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca
     4321 gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg
     4381 ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc
     4441 tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt
     4501 gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg
     4561 gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct
     4621 aagcacaatt cgagctcggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
     4681 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
     4741 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     4801 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     4861 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     4921 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa
     4981 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
     5041 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5101 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
     5161 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5221 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     5281 agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga
     5341 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     5401 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     5461 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc
     5521 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5581 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5641 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5701 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     5761 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     5821 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     5881 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5941 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     6001 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6061 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6121 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6181 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6241 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6301 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     6361 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     6421 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     6481 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     6541 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6601 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     6661 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6721 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     6781 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     6841 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     6901 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6961 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     7021 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     7081 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     7141 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     7201 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7261 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     7321 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     7381 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7441 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     7501 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     7561 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7621 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7681 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7741 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     7801 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     7861 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     7921 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     7981 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     8041 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     8101 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
     8161 agggaacaaa agctggagct gcaagctt
//
