LOCUS       sequence-347986-        8713 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 8713)
  TITLE     Improved vector system for knockdown
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8713)
  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            "Unpublished"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-SGIBX
FEATURES             Location/Qualifiers
     source          1..8713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     140..159
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             321..719
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    943..1531
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(996..1016)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1414..1425)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1738..1971
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(2072..2089)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2078..2189
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2400..2988)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2480..2499)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3159..3950,3951..4019))
                     /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     3782..3801
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4020..4124)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4196..4331
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4372..4506
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4409..4428)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4463..4482
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             4674..5307
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5354..5479
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5976..6209
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6394..6438
                     /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    6736..6853
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6902..6992
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     6913..6932
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     enhancer        7091..7394
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7395..7598
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7549..7569
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7595..7619
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      7728..7737
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7734..8432
                     /codon_start=1
                     /product="improved green fluorescent protein from 
                     Pontellina plumata (Evdokimov et al., 2006)"
                     /label=TurboGFP
                     /note="mammalian codon-optimized"
                     /translation="MESDESGLPAMEIECRITGTLNGVEFELVGGGEGTPEQGRMTNKM
                     KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL
                     HVSFSYRYEAGRVIGDFKVMGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNDLDGSFTR
                     TFSLRDGGYYSSVVDSHMHFKSAIHPSILQNGGPMFAFRRVEEDHSNTELGIVEYQHAF
                     KTPDADAGEE"
     misc_feature    join(8459..8713,1..318)
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8626..8643)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
ORIGIN
        1 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
       61 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
      121 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
      181 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
      241 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
      301 ttgaaaaaca cgataatacc atggccaagc ctttgtctca agaagaatcc accctcattg
      361 aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg
      421 cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg
      481 gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga
      541 cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc
      601 gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac
      661 agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctgat
      721 tcgaatgagg cttcagtact ttacagaatc gttgcctgca catcttggaa acacttgctg
      781 ggattacttc ttcaggttaa cccaacagaa ttctcttctg aagaaaatat atttcttttt
      841 attcatagct cttctcgagg taaccggatc caaccagttg tgttttaatg tatattaatg
      901 ttactaatgt gttttattta aattgagtca cgcgtctgga acaatcaacc tctggattac
      961 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     1021 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     1081 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     1141 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     1201 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     1261 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     1321 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg
     1381 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     1441 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     1501 agtcggatct ccctttgggc cgcctccccg cctggaatta attctgcagt cgagacctag
     1561 aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc
     1621 tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac
     1681 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag aggggactgg
     1741 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     1801 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     1861 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     1921 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     1981 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     2041 ggccttgaca ttgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     2101 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2161 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2221 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag cagggtatca
     2281 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2341 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2401 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2461 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2521 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2581 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2641 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2701 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2761 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2821 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2881 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2941 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3001 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3061 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3121 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3181 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3241 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3301 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3361 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3421 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3481 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3541 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3601 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3661 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3721 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3781 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3841 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3901 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3961 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4021 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4081 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4141 gtgccacctg acgtcgagtt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4201 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4261 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     4321 ttttggaggc ctaggctttt gcaaaaagct tcgtcgacgg atcgggagat caacttgttt
     4381 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4441 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4501 tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac cccataccct
     4561 attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta
     4621 ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt agttggaagg
     4681 gctaattcac tcccaaagaa gacaagatat ccttgatctg tggatctacc acacacaagg
     4741 ctacttccct gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt
     4801 tggatggtgc tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg
     4861 agagaacacc agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
     4921 agtgttagag tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca
     4981 tccggagtac ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga
     5041 ctttccaggg aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat
     5101 ataagcagct gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg
     5161 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     5221 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     5281 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
     5341 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
     5401 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
     5461 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
     5521 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     5581 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     5641 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     5701 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     5761 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     5821 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
     5881 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
     5941 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
     6001 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
     6061 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
     6121 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
     6181 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
     6241 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
     6301 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
     6361 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
     6421 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
     6481 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
     6541 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
     6601 ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac
     6661 agatccattc gattagtgaa cggatcggca ctgcgtgcgc caattctgca gacaaatggc
     6721 agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag
     6781 aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa
     6841 aattcaaaat tttcgggttt attacaggga cagcagagat ccagtttggt tagtactagt
     6901 aatatttgca tgtcgctatg tgttctggga aatcaccata aacgtgaaat gtctttggat
     6961 ttgggaatct tataagttct gtatgagacc acatcgattc tttttttcta gacgtattac
     7021 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     7081 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     7141 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     7201 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     7261 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     7321 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     7381 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     7441 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     7501 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     7561 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg
     7621 cctggagacg ccatccagcg ctgttttgac ctccatagaa gacaccggga ccgatccagc
     7681 ctccgcggcc gggaacggtg cattggaacg cggattcccc gtgtacagcc accatggaga
     7741 gcgacgagag cggcctgccc gccatggaga tcgagtgccg catcaccggc accctgaacg
     7801 gcgtggagtt cgagctggtg ggcggcggag agggcacccc cgagcagggc cgcatgacca
     7861 acaagatgaa gagcaccaaa ggcgccctga ccttcagccc ctacctgctg agccacgtga
     7921 tgggctacgg cttctaccac ttcggcacct accccagcgg ctacgagaac cccttcctgc
     7981 acgccatcaa caacggcggc tacaccaaca cccgcatcga gaagtacgag gacggcggcg
     8041 tgctgcacgt gagcttcagc taccgctacg aggccggccg cgtgatcggc gacttcaagg
     8101 tgatgggcac cggcttcccc gaggacagcg tgatcttcac cgacaagatc atccgcagca
     8161 acgccaccgt ggagcacctg caccccatgg gcgataacga tctggatggc agcttcaccc
     8221 gcaccttcag cctgcgcgac ggcggctact acagctccgt ggtggacagc cacatgcact
     8281 tcaagagcgc catccacccc agcatcctgc agaacggggg ccccatgttc gccttccgcc
     8341 gcgtggagga ggatcacagc aacaccgagc tgggcatcgt ggagtaccag cacgccttca
     8401 agaccccgga tgcagatgcc ggtgaagaat aatgtacaag tagcggccgc aaattccgcc
     8461 cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     8521 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     8581 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     8641 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     8701 caacgtctgt agc
//
