LOCUS       sequence-365629-        5259 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-3 vector for stable integration of up to three cassettes via 
            lentiviral transduction 
            (PCMV-5'LTR-Psi-gag-env-A1::A2::A3-WPRE-3'LTR).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5259)
  AUTHORS   Haellman V, Strittmatter T, Bertschi A, Stucheli P, Fussenegger M
  TITLE     A versatile plasmid architecture for mammalian synthetic biology 
            (VAMSyB).
  JOURNAL   Metab Eng. 2021 Jul;66:41-50. doi: 10.1016/j.ymben.2021.04.003. Epub
            2021 Apr 18.
  PUBMED    33857582
REFERENCE   2  (bases 1 to 5259)
  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"; journalName: "Metab 
            Eng."; date: "2021-07"; volume: "66:41-50. doi"; pages: " 
            10.1016/j.ymben.2021.04.003. Epub 2021 Apr 18"
COMMENT     Sequence Label: pTS1106_Tier3(Lenti)
FEATURES             Location/Qualifiers
     source          1..5259
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(29..820,821..889))
                     /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     652..671
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(890..994)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        1042..1345
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1347..1545
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1500..1520
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1563..1743
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1790..1915
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2408..2641
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2826..2870
                     /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    3168..3285
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3623..4211
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3676..3696)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4094..4105)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4283..4516
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     rep_origin      complement(4579..5167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4659..4678)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
       61 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
      121 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcgg ctcccacgct
      181 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
      241 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
      301 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
      361 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
      421 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
      481 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
      541 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
      601 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
      661 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
      721 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
      781 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
      841 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
      901 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
      961 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     1021 acgtctaaga aaccaagatc tcgttacata acttacggta aatggcccgc ctggctgacc
     1081 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     1141 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     1201 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     1261 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     1321 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     1381 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     1441 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     1501 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagcgcgt tttgcctgta
     1561 ctgggtttct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     1621 cactgcttaa gcctcaataa agtttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     1681 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     1741 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg
     1801 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
     1861 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat
     1921 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa
     1981 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa
     2041 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc
     2101 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg
     2161 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga
     2221 gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag
     2281 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     2341 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     2401 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     2461 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     2521 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     2581 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     2641 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     2701 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     2761 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
     2821 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
     2881 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     2941 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
     3001 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa
     3061 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg
     3121 cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa
     3181 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
     3241 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
     3301 gacagcagtg acgcgagaag aagcggggat acttggtcac gcgtggtacc ctcgaggagc
     3361 tcttcgaagc ggaattcacc atgactagtg ctagcggatc caccggtgtc tagaaagctt
     3421 tgaggccggc ctgcaggggt accggatttc ttgtggacac aacactctcg agcgcgggag
     3481 accgtcagtt ggtctccagc tctcatagct gtttcctgtg taaaacgacg gccagcgcgg
     3541 gagacggtca gttcgtctcc agctattaat tgcgttgcgc tcactgcccg ctttccagtc
     3601 gggaaacctg tcgtgccaga tcaatcaacc tctggattac aaaatttgtg aaagattgac
     3661 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3721 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3781 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3841 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3901 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3961 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     4021 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     4081 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4141 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4201 cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     4261 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
     4321 ctttttgctt gtactgggtt tctctggtta gaccagatct gagcctggga gctctctggc
     4381 taactaggga acccactgct taagcctcaa taaagtttgc cttgagtgct tcaagtagtg
     4441 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     4501 tggaaaatct ctagcagaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4561 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4621 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4681 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4741 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4801 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4861 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4921 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4981 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5041 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5101 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5161 atctcaagaa gatcctttga tcttttctac ggggattatc aaaaaggatc ttcacctaga
     5221 tccttttaaa ttaaaaatga agttttaaat caatctaaa
//
