LOCUS       sequence-259352-        6118 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivector for CypA knockdown and blasticidin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6118)
  AUTHORS   Kim K, Dauphin A, Komurlu S, McCauley SM, Yurkovetskiy L, Carbone C,
            Diehl WE, Strambio-De-Castillia C, Campbell EM, Luban J
  TITLE     Cyclophilin A protects HIV-1 from restriction by human TRIM5alpha.
  JOURNAL   Nat Microbiol. 2019 Dec;4(12):2044-2051. doi: 
            10.1038/s41564-019-0592-5. Epub 2019 Oct 21.
  PUBMED    31636416
REFERENCE   2  (bases 1 to 6118)
  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"; doi: 
            "10.1038/s41564-019-0592-5"; journalName: "Nat Microbiol"; date: 
            "2019-12"; volume: "4"; issue: "12"; pages: "2044-2051"
COMMENT     Sequence Label: pABM-D4-miR30-CypA
FEATURES             Location/Qualifiers
     source          1..6118
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(59..79)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        64..393
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      244..379
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     306..325
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        405..784
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        786..984
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     939..959
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1002..1182
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1229..1354
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1851..2084
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2269..2313
                     /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    2611..2728
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2865..3272
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      3287..3296
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3477..3488
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     ncRNA           3786..3800
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     LTR             3934..4114
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4185..4773)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4265..4284)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4944..5735,5736..5804))
                     /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     5567..5586
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5805..5909)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5984..6003)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgctgtgg
       61 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
      121 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
      181 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
      241 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
      301 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
      361 ggaggctttt ttggaggcct aggcttttgc aaaaagctcg cgttgacatt gattattgac
      421 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      481 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      541 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      601 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      661 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      721 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      781 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      841 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      901 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      961 acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc
     1021 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     1081 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     1141 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
     1201 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
     1261 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
     1321 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
     1381 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
     1441 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
     1501 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     1561 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     1621 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
     1681 cgcacagcaa gcggccggcc gctgatcttc agacctggag gaggagatat gagggacaat
     1741 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     1801 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     1861 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     1921 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     1981 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     2041 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     2101 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     2161 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     2221 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     2281 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     2341 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     2401 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     2461 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
     2521 gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt
     2581 ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta
     2641 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
     2701 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt
     2761 ttggttaatt aactgcagcc ccgataaaat aaaagatttt atttagtctc cagaaaaagg
     2821 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc tgcagtaacg ccattttgca
     2881 aggcatggaa aaataccaaa ccaagaatag agaagttcag atcaagggcg ggtacatgaa
     2941 aatagctaac gttgggccaa acaggatatc tgcggtgagc agtttcggcc ccggcccggg
     3001 gccaagaaca gatggtcacc gcagtttcgg ccccggcccg aggccaagaa cagatggtcc
     3061 ccagatatgg cccaaccctc agcagtttct taagacccat cagatgtttc caggctcccc
     3121 caaggacctg aaatgaccct gcgccttatt tgaattaacc aatcagcctg cttctcgctt
     3181 ctgttcgcgc gcttctgctt cccgagctct ataaaagagc tcacaacccc tcactcggcg
     3241 cgccagtcct ccgacagact gagtcgcccg ggggtctaga acgcgtgccg ccatggctaa
     3301 accactgtcc caggaagaat ccacactgat cgaacgagca accgctacta tcaactcaat
     3361 cccaatctcc gaagactact ctgtggcctc cgccgccctg agcagcgacg gacgaatctt
     3421 caccggcgtg aacgtctacc actttaccgg cggcccctgc gctgagctgg tggtcctggg
     3481 aactgcagcc gctgcagccg ctggaaatct gacctgtatc gtggctatcg ggaacgaaaa
     3541 taggggaatc ctgtctccat gcggccggtg tagacaggtc ctgctggatc tgcatcccgg
     3601 gatcaaggca attgtgaaag actcagatgg acagcccaca gccgtcggca ttcgcgagct
     3661 gctgcctagc ggctatgtgt gggaagggtg agcgatcgcc gcggccgcct tcttaaccca
     3721 acagaaggct cgagaaggta tattgctgtt gacagtgagc gatggattgc agagttaagt
     3781 ttatagtgaa gccacagatg tataaactta actctgcaat ccagtgccta ctgcctcgga
     3841 cttcaagggg ctagaattcg gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     3901 gactggaagg gctaactgca tccggactgt actgggtctc tctggttaga ccagatctga
     3961 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     4021 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     4081 agaccctttt agtcagtgtg gaaaatctct agcagggccc gtttcatgtg agcaaaaggc
     4141 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4201 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4261 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4321 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4381 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4441 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4501 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4561 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4621 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4681 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4741 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4801 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4861 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4921 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4981 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5041 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5101 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5161 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5221 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5281 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5341 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5401 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5461 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5521 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5581 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5641 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5701 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5761 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5821 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5881 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     5941 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     6001 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     6061 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatct
//
