LOCUS       sequence-344690-        8788 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 8788)
  TITLE     Improved vector system for knockdown
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8788)
  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            "Unpublished"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-U6-SIPZ
FEATURES             Location/Qualifiers
     source          1..8788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     136..155
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             317..919
                     /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     816..836
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    1144..1732
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1197..1217)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1615..1626)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1939..2172
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(2273..2290)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2279..2390
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2601..3189)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2681..2700)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3360..4151,4152..4220))
                     /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     3983..4002
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4221..4325)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4397..4532
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4573..4707
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4610..4629)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4664..4683
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             4875..5508
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5555..5680
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6177..6410
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6595..6639
                     /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    6937..7054
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(7119..7493)
                     /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"
     regulatory      7490..7499
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7503..7521)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(7513..7560)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        7587..7827
                     /label=U6 promoter
                     /note="Truncated RNA polymerase III promoter for human U6 
                     snRNA used in pSIREN vectors such as 
                     pSIREN-RetroQ-ZsGreen1. Expected transcription start site 
                     is 8bp downstream of this feature."
     primer_bind     7587..7607
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7758..7777
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        7926..8229
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8230..8433
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8384..8404
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8430..8454
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    join(8530..8788,1..314)
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8697..8714)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
ORIGIN
        1 ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt
       61 gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt
      121 gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag
      181 aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt
      241 tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga
      301 aaaacacgat aataccatgg ccaccgagta caagcccacg gtgcgcctcg ccacccgcga
      361 cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
      421 ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
      481 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
      541 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggctcgcg
      601 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
      661 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
      721 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
      781 tggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
      841 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
      901 ccgcaagccc ggtgcctgag tttgtttgaa tgaggcttca gtactttaca gaatcgttgc
      961 ctgcacatct tggaaacact tgctgggatt acttcttcag gttaacccaa cagaattctc
     1021 ttctgaagaa aatatatttc tttttattca tagctcttct cgaggtaacc ggatccagtt
     1081 gtgttttaat gtatattaat gttactaatg tgttttattt aaattgagtc acgcgtctgg
     1141 aacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     1201 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     1261 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     1321 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     1381 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     1441 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     1501 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     1561 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     1621 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     1681 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt
     1741 aattctgcag tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta
     1801 ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca
     1861 cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     1921 taaaagaaaa gaggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt
     1981 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     2041 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     2101 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     2161 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     2221 aatgaatatc agagagtgag aggccttgac attgtttaaa cccgctgatc agcctcgact
     2281 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2341 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2401 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2461 gaagacaata gcagggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2521 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2581 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2641 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2701 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2761 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2821 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2881 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2941 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3001 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3061 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3121 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3181 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3241 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3301 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3361 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3421 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3481 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3541 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3601 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3661 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3721 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3781 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3841 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3901 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3961 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4021 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4081 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4141 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4201 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4261 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4321 ccgcgcacat ttccccgaaa agtgccacct gacgtcgagt tagtcagcaa ccatagtccc
     4381 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4441 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4501 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttcgtcgacg
     4561 gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4621 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4681 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc
     4741 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa
     4801 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact
     4861 acaaacttag tagttggaag ggctaattca ctcccaaaga agacaagata tccttgatct
     4921 gtggatctac cacacacaag gctacttccc tgattagcag aactacacac cagggccagg
     4981 ggtcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg agccagataa
     5041 ggtagaagag gccaataaag gagagaacac cagcttgtta caccctgtga gcctgcatgg
     5101 gatggatgac ccggagagag aagtgttaga gtggaggttt gacagccgcc tagcatttca
     5161 tcacgtggcc cgagagctgc atccggagta cttcaagaac tgctgatatc gagcttgcta
     5221 caagggactt tccgctgggg actttccagg gaggcgtggc ctgggcggga ctggggagtg
     5281 gcgagccctc agatcctgca tataagcagc tgctttttgc ctgtactggg tctctctggt
     5341 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     5401 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     5461 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
     5521 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
     5581 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
     5641 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
     5701 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
     5761 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
     5821 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
     5881 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
     5941 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
     6001 gaccaccgca cagcaagcgg ccggccgctg atcttcagac ctggaggagg agatatgagg
     6061 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     6121 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     6181 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     6241 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     6301 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     6361 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     6421 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     6481 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     6541 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     6601 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     6661 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     6721 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     6781 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa
     6841 ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg
     6901 ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg
     6961 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
     7021 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga
     7081 tccagtttgg ttagtaccgg gcccgctcta gtccggaatc agtcctgctc ctcggccacg
     7141 aagtgcacgc agttgccggc cgggtcgcgc agggcgaact cccgccccca cggctgctcg
     7201 ccgatctcgg tcatggccgg cccggaggcg tcccggaagt tcgtggacac gacctccgac
     7261 cactcggcgt acagctcgtc caggccgcgc acccacaccc aggccagggt gttgtccggc
     7321 accacctggt cctggaccgc gctgatgaac agggtcacgt cgtcccggac cacaccggcg
     7381 aagtcgtcct ccacgaagtc ccgggagaac ccgagccggt cggtccagaa ctcgaccgct
     7441 ccggcgacgt cgcgcgcggt gagcaccgga acggcactgg tcaacttggc catggtggcc
     7501 ctcctatagt gagtcgtatt atactatgcc gatatactat gccgatgatt aattgtcaac
     7561 acgtgctgca ggtccgaggt tctagtgagg gcctatttcc catgattcct tcatatttgc
     7621 atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga
     7681 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
     7741 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
     7801 tggctttata tatcttgtgg aaaggacatc gattcttttt ttctagacgt attaccgcca
     7861 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     7921 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     7981 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     8041 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     8101 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     8161 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     8221 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     8281 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     8341 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     8401 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
     8461 agacgccatc cacgctgttt tgacctccat agaagacacc gactgtacaa gtagcggccg
     8521 caaattccgc ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata
     8581 aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt
     8641 gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct
     8701 cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc
     8761 ttgaagacaa acaacgtctg tagcgacc
//
