LOCUS       sequence-344693-        8923 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 8923)
  TITLE     Improved vector system for knockdown
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8923)
  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            "Unpublished"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-SGIPX
FEATURES             Location/Qualifiers
     source          1..8923
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     76..93
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(287..985)
                     /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"
     regulatory      982..991
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1100..1124)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(1121..1324)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1150..1170)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1325..1628
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        complement(1727..1817)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     complement(1787..1806)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_feature    1866..1983
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(2281..2325)
                     /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    2510..2743
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    3240..3365
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             3412..4045
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     polyA_signal    4213..4347
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4237..4256)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4291..4310
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4388..4523
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4595..4699
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4700..4768,4769..5560)
                     /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     complement(4918..4937)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5731..6319
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6220..6239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6530..6641
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6630..6647
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             6748..6981
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    7188..7776
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             7294..7305
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     7703..7723
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8006..8608)
                     /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     complement(8089..8109)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    join(8611..8923,1..260)
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8770..8789)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
ORIGIN
        1 gggtcgctac agacgttgtt tgtcttcaag aagcttccag aggaactgct tccttcacga
       61 cattcaacag accttgcatt cctttggcga gaggggaaag acccctagga atgctcgtca
      121 agaagacagg gccaggtttc cgggccctca cattgccaaa agacggcaat atggtggaaa
      181 ataacatata gacaaacgca caccggcctt attccaagcg gcttcggcca gtaacgttag
      241 ggggggggga gggagagggg cggaatttgc ggccgctact tgtacattat tcttcaccgg
      301 catctgcatc cggggtcttg aaggcgtgct ggtactccac gatgcccagc tcggtgttgc
      361 tgtgatcctc ctccacgcgg cggaaggcga acatggggcc cccgttctgc aggatgctgg
      421 ggtggatggc gctcttgaag tgcatgtggc tgtccaccac ggagctgtag tagccgccgt
      481 cgcgcaggct gaaggtgcgg gtgaagctgc catccagatc gttatcgccc atggggtgca
      541 ggtgctccac ggtggcgttg ctgcggatga tcttgtcggt gaagatcacg ctgtcctcgg
      601 ggaagccggt gcccatcacc ttgaagtcgc cgatcacgcg gccggcctcg tagcggtagc
      661 tgaagctcac gtgcagcacg ccgccgtcct cgtacttctc gatgcgggtg ttggtgtagc
      721 cgccgttgtt gatggcgtgc aggaaggggt tctcgtagcc gctggggtag gtgccgaagt
      781 ggtagaagcc gtagcccatc acgtggctca gcaggtaggg gctgaaggtc agggcgcctt
      841 tggtgctctt catcttgttg gtcatgcggc cctgctcggg ggtgccctct ccgccgccca
      901 ccagctcgaa ctccacgccg ttcagggtgc cggtgatgcg gcactcgatc tccatggcgg
      961 gcaggccgct ctcgtcgctc tccatggtgg ctgtacacgg ggaatccgcg ttccaatgca
     1021 ccgttcccgg ccgcggaggc tggatcggtc ccggtgtctt ctatggaggt caaaacagcg
     1081 ctggatggcg tctccaggcg atctgacggt tcactaaacg agctctgctt atatagacct
     1141 cccaccgtac acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg
     1201 gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac
     1261 ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca
     1321 tcaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata
     1381 aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg
     1441 gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact
     1501 ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta
     1561 ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt
     1621 atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt
     1681 aataactaat gcatggcggt aatacgtcta gaaaaaaaga atcgatgtgg tctcatacag
     1741 aacttataag attcccaaat ccaaagacat ttcacgttta tggtgatttc ccagaacaca
     1801 tagcgacatg caaatattac tagtactaac caaactggat ctctgctgtc cctgtaataa
     1861 acccgaaaat tttgaatttt tgtaatttgt ttttgtaatt ctttagtttg tatgtctgtt
     1921 gctattatgt ctactattct ttcccctgca ctgtaccccc caatcccccc ttttctttta
     1981 aaattgtgga tgaatactgc catttgtctg cagaattggc gcacgcagtg ccgatccgtt
     2041 cactaatcga atggatctgt ctctgtctct ctctccacct tcttcttcta ttccttcggg
     2101 cctgtcgggt cccctcgggg ttgggaggtg ggtctgaaac gataatggtg aatatccctg
     2161 cctaactcta ttcactatag aaagtacagc aaaaactatt cttaaaccta ccaagcctcc
     2221 tactatcatt atgaataatt ttatatacca cagccaattt gttatgttaa accaattcca
     2281 caaacttgcc catttatcta attccaataa ttcttgttca ttcttttctt gctggttttg
     2341 cgattcttca attaaggagt gtattaagct tgtgtaattg ttaatttctc tgtcccactc
     2401 catccaggtc gtgtgattcc aaatctgttc cagagattta ttactccaac tagcattcca
     2461 aggcacagca gtggtgcaaa tgagttttcc agagcaaccc caaatcccca ggagctgttg
     2521 atcctttagg tatctttcca cagccaggat tcttgcctgg agctgcttga tgccccagac
     2581 tgtgagttgc aacagatgct gttgcgcctc aatagccctc agcaaattgt tctgctgctg
     2641 cactatacca gacaataatt gtctggcctg taccgtcagc gtcattgacg ctgcgcccat
     2701 agtgcttcct gctgctccca agaacccaag gaacaaagct cctattccca ctgctctttt
     2761 ttctctctgc accactcttc tctttgcctt ggtgggtgct actcctaatg gttcaatttt
     2821 tactacttta tatttatata attcacttct ccaattgtcc ctcatatctc ctcctccagg
     2881 tctgaagatc agcggccggc cgcttgctgt gcggtggtct tacttttgtt ttgctcttcc
     2941 tctatcttgt ctaaagcttc cttggtgtct tttatctcta tcctttgatg cacacaatag
     3001 agggttgcta ctgtattata taatgatcta agttcttctg atcctgtctg aagggatggt
     3061 tgtagctgtc ccagtatttg tctacagcct tctgatgttt ctaacaggcc aggattaact
     3121 gcgaatcgtt ctagctccct gcttgcccat actatatgtt ttaatttata ttttttcttt
     3181 ccccctggcc ttaaccgaat tttttcccat cgcgatctaa ttctcccccg cttaatactg
     3241 acgctctcgc acccatctct ctccttctag cctccgctag tcaaaatttt tggcgtactc
     3301 accagtcgcc gcccctcgcc tcttgccgtg cgcgcttcag caagccgagt cctgcgtcga
     3361 gagagctcct ctggtttccc tttcgctttc aagtccctgt tcgggcgcca ctgctagaga
     3421 ttttccacac tgactaaaag ggtctgaggg atctctagtt accagagtca cacaacagac
     3481 gggcacacac tacttgaagc actcaaggca agctttattg aggcttaagc agtgggttcc
     3541 ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac ccagtacagg
     3601 caaaaagcag ctgcttatat gcaggatctg agggctcgcc actccccagt cccgcccagg
     3661 ccacgcctcc ctggaaagtc cccagcggaa agtcccttgt agcaagctcg atatcagcag
     3721 ttcttgaagt actccggatg cagctctcgg gccacgtgat gaaatgctag gcggctgtca
     3781 aacctccact ctaacacttc tctctccggg tcatccatcc catgcaggct cacagggtgt
     3841 aacaagctgg tgttctctcc tttattggcc tcttctacct tatctggctc aactggtact
     3901 agcttgtagc accatccaaa ggtcagtgga tatctgaccc ctggccctgg tgtgtagttc
     3961 tgctaatcag ggaagtagcc ttgtgtgtgg tagatccaca gatcaaggat atcttgtctt
     4021 ctttgggagt gaattagccc ttccaactac taagtttgta gtacatattt aacaaataca
     4081 atttctttaa aatgaaaata attcagagga atcacaggtt tagagtaaat gaaaccacag
     4141 gtaattggca gtggtaatag ggtatggggt gggaagtttg ggatgatttt ggttagcttg
     4201 agttatccag ttgatccaga catgataaga tacattgatg agtttggaca aaccacaact
     4261 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     4321 accattataa gctgcaataa acaagttgat ctcccgatcc gtcgacgaag ctttttgcaa
     4381 aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg
     4441 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
     4501 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
     4561 ctcgacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4621 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4681 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4741 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4801 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4861 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4921 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4981 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     5041 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5101 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5161 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5221 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5281 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5341 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5401 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5461 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5521 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5581 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5641 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5701 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5761 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5821 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5881 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5941 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6001 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6061 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6121 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6181 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6241 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6301 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6361 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6421 taccgccttt gagtgagctg ataccctgct attgtcttcc caatcctccc ccttgctgtc
     6481 ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc
     6541 ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg
     6601 ggaggggcaa acaacagatg gctggcaact agaaggcaca gtcgaggctg atcagcgggt
     6661 ttaaacaatg tcaaggcctc tcactctctg atattcattt ctttgcaagt tataaatact
     6721 gaataataag atgacatgaa ctactactgc tagagatttt ccacactgac taaaagggtc
     6781 tgagggatct ctagttacca gagtcacaca acagacgggc acacactact tgaagcactc
     6841 aaggcaagct ttattgaggc ttaagcagtg ggttccctag ttagccagag agctcccagg
     6901 ctcagatctg gtctaaccag agagacccag tacaagcaaa aagcagatct tgtcttcgtt
     6961 gggagtgaat tagcccttcc agtcccctct tttcttttaa aaagtggcta agatctacag
     7021 ctgccttgta agtcattggt cttaaaggta cctgaggtgt gactggaaaa cccacctcct
     7081 cctcctcttg tgcttctagc caggcacaat cagcattggt agctgctgta ttgctacttg
     7141 tgattgctcc atgtttttct aggtctcgac tgcagaatta attccaggcg gggaggcggc
     7201 ccaaagggag atccgactcg tctgagggcg aaggcgaaga cgcggaagag gccgcagagc
     7261 cggcagcagg ccgcgggaag gaaggtccgc tggattgagg gccgaaggga cgtagcagaa
     7321 ggacgtcccg cgcagaatcc aggtggcaac acaggcgagc agccatggaa aggacgtcag
     7381 cttccccgac aacaccacgg aattgtcagt gcccaacagc cgagcccctg tccagcagcg
     7441 ggcaaggcag gcggcgatga gttccgccgt ggcaataggg agggggaaag cgaaagtccc
     7501 ggaaaggagc tgacaggtgg tggcaatgcc ccaaccagtg ggggttgcgt cagcaaacac
     7561 agtgcacacc acgccacgtt gcctgacaac gggccacaac tcctcataaa gagacagcaa
     7621 ccaggattta tacaaggagg agaaaatgaa agccatacgg gaagcaatag catgatacaa
     7681 aggcattaaa gcagcgtatc cacatagcgt aaaaggagca acatagttaa gaataccagt
     7741 caatctttca caaattttgt aatccagagg ttgattgttc cagacgcgtg actcaattta
     7801 aataaaacac attagtaaca ttaatataca ttaaaacaca actggttgga tccggttacc
     7861 tcgagaagag ctatgaataa aaagaaatat attttcttca gaagagaatt ctgttgggtt
     7921 aacctgaaga agtaatccca gcaagtgttt ccaagatgtg caggcaacga ttctgtaaag
     7981 tactgaagcc tcattcgaaa caaactcagg caccgggctt gcgggtcatg caccaggtgc
     8041 gcggtccttc gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc tcgtagaagg
     8101 ggaggttgcg gggcgcggag gtctccagga aggcgggcac cccagcgcgc tcggccgcct
     8161 ccactccggg gagcacgacg gcgctgccca gacccttgcc ctggtggtcg ggcgagacgc
     8221 cgacggtggc caggaaccac gcgggctcct tgggccggtg cggcgccagg aggccttcca
     8281 tctgttgctg cgcggccagc cgggaaccgc tcaactcggc catgcgcgag ccgatctcgg
     8341 cgaacaccgc ccccgcttcg acgctctccg gcgtggtcca gaccgccacc gcggcgccgt
     8401 cgtccgcgac ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag agttcttgca
     8461 gctcggtgac ccgctcgatg tggcggtccg ggtcgacggt gtggcgcgtg gcggggtagt
     8521 cggcgaacgc ggcggcgagg gtgcgtacgg cccgggggac gtcgtcgcgg gtggcgaggc
     8581 gcaccgtggg cttgtactcg gtggccatgg tattatcgtg tttttcaaag gaaaaccacg
     8641 tccccgtggt tcggggggcc tagacgtttt tttaacctcg actaaacaca tgtaaagcat
     8701 gtgcaccgag gccccagatc agatcccata caatggggta ccttctgggc atccttcagc
     8761 cccttgttga atacgcttga ggagagccat ttgactcttt ccacaactat ccaactcaca
     8821 acgtggcact ggggttgtgc cgcctttgca ggtgtatctt atacacgtgg cttttggccg
     8881 cagaggcacc tgtcgccagg tggggggttc cgctgcctgc aaa
//
