LOCUS       sequence-344691-        8212 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  empty vector for cloning shRNA and/or miRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8212)
  TITLE     Improved vector system for knockdown
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8212)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pLV-H1-SIPX
FEATURES             Location/Qualifiers
     source          1..8212
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     7..31
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    160..732
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(327..344)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     554..573
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             735..1337
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     1234..1254
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    1567..2155
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1620..1640)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2038..2049)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2362..2595
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(2696..2713)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2702..2813
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3024..3612)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3104..3123)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3783..4574,4575..4643))
                     /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     4406..4425
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4644..4748)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4820..4955
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4996..5130
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5033..5052)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5087..5106
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             5298..5931
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5978..6103
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6600..6833
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7018..7062
                     /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    7360..7477
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7526..7616
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     7537..7556
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     enhancer        7715..8018
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(8019..8212,1..10)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8173..8193
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccag cgctgttttg
       61 acctccatag aagacaccgg gaccgatcca gcctccgcgg ccgggaacgg tgcattggaa
      121 cgcggattcc ccgtgtacaa gtagcggccg caaattccgc ccctctccct cccccccccc
      181 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
      241 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
      301 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
      361 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
      421 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
      481 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
      541 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
      601 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
      661 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
      721 aacacgataa taccatggcc accgagtaca agcccacggt gcgcctcgcc acccgcgacg
      781 acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
      841 acaccgtcga cccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
      901 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
      961 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggctcgcgca
     1021 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     1081 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
     1141 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgctg
     1201 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     1261 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     1321 gcaagcccgg tgcctgagtt tgtttcgaat gaggcttcag tactttacag aatcgttgcc
     1381 tgcacatctt ggaaacactt gctgggatta cttcttcagg ttaacccaac agaattctct
     1441 tctgaagaaa atatatttct ttttattcat agctcttctc gaggtaaccg gatccaacca
     1501 gttgtgtttt aatgtatatt aatgttacta atgtgtttta tttaaattga gtcacgcgtc
     1561 tggaacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     1621 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     1681 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     1741 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     1801 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     1861 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     1921 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     1981 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     2041 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     2101 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga
     2161 attaattctg cagtcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag
     2221 ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag
     2281 tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     2341 ttttaaaaga aaagagggga ctggaagggc taattcactc ccaacgaaga caagatctgc
     2401 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     2461 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     2521 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     2581 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa
     2641 agaaatgaat atcagagagt gagaggcctt gacattgttt aaacccgctg atcagcctcg
     2701 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2761 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2821 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2881 tgggaagaca atagcagggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     2941 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3001 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3061 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3121 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3181 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3241 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3301 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3361 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3421 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3481 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3541 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3601 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3661 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3721 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3781 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3841 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3901 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3961 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4021 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4081 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4141 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4201 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4261 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4321 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4381 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4441 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4501 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4561 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4621 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4681 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     4741 gttccgcgca catttccccg aaaagtgcca cctgacgtcg agttagtcag caaccatagt
     4801 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4861 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4921 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttcgtcg
     4981 acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5041 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5101 ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat
     5161 cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc
     5221 taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt
     5281 actacaaact tagtagttgg aagggctaat tcactcccaa agaagacaag atatccttga
     5341 tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca caccagggcc
     5401 aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga
     5461 taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca
     5521 tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt
     5581 tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg
     5641 ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga
     5701 gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct
     5761 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     5821 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     5881 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
     5941 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
     6001 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
     6061 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
     6121 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
     6181 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
     6241 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
     6301 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
     6361 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
     6421 taagaccacc gcacagcaag cggccggccg ctgatcttca gacctggagg aggagatatg
     6481 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     6541 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     6601 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     6661 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     6721 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     6781 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     6841 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     6901 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     6961 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     7021 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     7081 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     7141 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     7201 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     7261 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     7321 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     7381 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     7441 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     7501 agatccagtt tggttagtac tagtaatatt tgcatgtcgc tatgtgttct gggaaatcac
     7561 cataaacgtg aaatgtcttt ggatttggga atcttataag ttctgtatga gaccacatcg
     7621 attctttttt tctagacgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg
     7681 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     7741 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     7801 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     7861 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     7921 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     7981 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     8041 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     8101 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     8161 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat at
//
