LOCUS       sequence-409604-        8279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty lentiviral vector to express protein in mammalian cells 
            cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8279)
  AUTHORS   Sakai E, Imaizumi T, Suzuki R, Taracena-Gandara M, Fujimoto T, 
            Sakurai F, Mizuguchi H
  TITLE     miR-27b targets MAIP1 to mediate lipid accumulation in cultured 
            human and mouse hepatic cells.
  JOURNAL   Commun Biol. 2023 Jun 24;6(1):669. doi: 10.1038/s42003-023-05049-w.
  PUBMED    37355744
REFERENCE   2  (bases 1 to 8279)
  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: "Commun 
            Biol."; date: "2023-06-24"; volume: "6(1):669. doi"; pages: " 
            10.1038/s42003-023-05049-w"
COMMENT     Sequence Label: CSII-TK-MCS
FEATURES             Location/Qualifiers
     source          1..8279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        616..814
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             832..1012
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1059..1184
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1676..1909
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2094..2138
                     /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    2412..2529
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2740..2884
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     misc_feature    2974..3562
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3027..3047)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3445..3456)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4081..4261
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4287..4304)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4293..4517
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4563..4991
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4650..4669)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4860..4881
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5000..5020)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5005..5334
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5185..5320
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5247..5266
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5382..5429
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5448..5822
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5952..6073
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5989..6008)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6043..6062
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6122..6138)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6122..6138)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6135..6157)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6146..6162
                     /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)."
     promoter        complement(join(6170..6176,6177..6194,6195..6200))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6215..6236
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6353..6370)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6524..7112)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6604..6623)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7283..8074,8075..8143))
                     /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     7906..7925
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8144..8248)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc
      841 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      901 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      961 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     1021 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     1081 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     1141 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     1201 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     1261 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     1321 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     1381 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     1441 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     1501 gtaagaccac cgcacagcaa gggccgctga tcttcagacc tggaggagga gatatgaggg
     1561 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1621 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1681 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     1741 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1801 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1861 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1921 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1981 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     2041 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     2101 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     2161 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     2221 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     2281 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
     2341 gtggagagag agacagagac agatccattc gattagtgaa cggatctaca aatggcagta
     2401 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     2461 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     2521 caaaattttc gggtttatta cagggacagc agaaattcac tttgaattaa ttcaagcttc
     2581 gtgaggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     2641 ggggggaggg gtcggcaatt gaaccggtcc gtggcccctt gctcgcgttt gctggcggtg
     2701 tccccggaag aaatatattt gcatgtcttt agttctatga tgacacaaac cccgcccagc
     2761 gtcttgtcat tggcgaattc gaacacgcag atgcagtcgg ggcggcgcgg tcccaggtcc
     2821 acttcgcata ttaaggtgac gcgtgtggcc tcgaacaccg agcgaccctg cagcgacccg
     2881 cttagcggcc gcgggtcata aaaatggcgc tggaaaaatc agcttatgga ctggatcctc
     2941 tagagttaac atcgagggat caagcttatc gataatcaac ctctggatta caaaatttgt
     3001 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3061 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3121 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     3181 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     3241 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     3301 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     3361 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     3421 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     3481 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     3541 tccctttggg ccgcctcccc gcatcgatac cgtcgagacc tagaaaaaca tggagcaatc
     3601 acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag
     3661 gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca
     3721 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     3781 caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat
     3841 tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg atggtgctac
     3901 aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga gaacacccgc
     3961 ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt attagagtgg
     4021 aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc ggactgtact
     4081 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     4141 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     4201 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     4261 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     4321 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     4381 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     4441 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     4501 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     4561 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4621 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4681 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4741 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4801 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4861 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4921 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4981 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     5041 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     5101 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5161 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5221 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     5281 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     5341 ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg
     5401 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga
     5461 ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg
     5521 accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg
     5581 acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct
     5641 gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga
     5701 acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg
     5761 agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact
     5821 gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5881 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5941 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6001 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6061 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     6121 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6181 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6241 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6301 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6361 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6421 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6481 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6541 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6601 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6661 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6721 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6781 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6841 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6901 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6961 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7021 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7081 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7141 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7201 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7261 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7321 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7381 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7441 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7501 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7561 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7621 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7681 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7741 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7801 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7861 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7921 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7981 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8041 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8101 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8161 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8221 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
