LOCUS       sequence-246619-        5870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses shRNA against human CNP under mouse U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5870)
  AUTHORS   Unlu I, Lu Y, Wang X
  TITLE     The cyclic phosphodiesterase CNP and RNA cyclase RtcA fine-tune 
            noncanonical XBP1 splicing during ER stress.
  JOURNAL   J Biol Chem. 2018 Dec 14;293(50):19365-19376. doi: 
            10.1074/jbc.RA118.004872. Epub 2018 Oct 24.
  PUBMED    30355738
REFERENCE   2  (bases 1 to 5870)
  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"; doi: 
            "10.1074/jbc.RA118.004872"; journalName: "J Biol Chem"; date: 
            "2018-12-14- 14"; volume: "293"; issue: "50"; pages: "19365-19376"
COMMENT     Sequence Label: pB-puro-mU6-CNPshRNA1
FEATURES             Location/Qualifiers
     source          1..5870
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      3..12
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(26..525)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(74..93)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     467..490
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     promoter        574..887
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     806..826
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     repeat_region   1291..1353
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(1419..1438)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1420..1438)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1445..1462)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1445..1461)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1454..1476)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1603..2058
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1690..1709)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1900..1921
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2084..2188
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2189..2257,2258..3049)
                     /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(2407..2426)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3220..3808
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3709..3728
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3962..3979
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4096..4117
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4132..4137,4138..4155,4156..4162)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4170..4186
                     /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)."
     primer_bind     4175..4197
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4194..4210
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4194..4210
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4231..4249
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   4398..4432
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    4819..4940
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4830..4849)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4884..4903
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5274..5870,1..6))
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5357..5377)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 ccccatggtg gcgttggctg caggtcgaaa ggcccggaga tgaggaagag gagaacagcg
       61 cggcagacgt gcgcttttga agcgtgcaga atgccgggcc tccggaggac cttcgggcgc
      121 ccgccccgcc cctgagcccg cccctgagcc cgcccccgga cccacccctt cccagcctct
      181 gagcccagaa agcgaaggag caaagctgct attggccgct gccccaaagg cctacccgct
      241 tccattgctc agcggtgctg tccatctgca cgagactagt gagacgtgct acttccattt
      301 gtcacgtcct gcacgacgcg agctgcgggg cgggggggaa cttcctgact aggggaggag
      361 tagaaggtgg cgcgaagggg ccaccaaaga acggagccgg ttggcgccta ccggtggatg
      421 tggaatgtgt gcgaggccag aggccacttg tgtagcgcca agtgcccagc ggggctgcta
      481 aagcgcatgc tccagactgc cttgggaaaa gcgcctcccc tacccggtag aattaattcg
      541 atggccgcgg tccacactcg ccccaacgcc cgggatccga cgccgccatc tctaggcccg
      601 cgccggcccc ctcgcacaga cttgtgggag aagctcggct actcccctgc cccggttaat
      661 ttgcatataa tatttcctag taactataga ggcttaatgt gcgataaaag acagataatc
      721 tgttcttttt aatactagct acattttaca tgataggctt ggatttctat aagagataca
      781 aatactaaat tattatttta aaaaacagca caaaaggaaa ctcaccctaa ctgtaaagta
      841 attgtgtgtt ttgagactat aaatatccct tggagaaaag ccttgtttgt ggagttaagc
      901 gagcagcaac tagagttgct gctcgcttaa ctccactttt tctcgagtta attaacgaga
      961 gcataatatt gatatgtgcc aaagttgttt ctgactgact aataagtata atttgtttct
     1021 attatgtata ggttaagcta attacttatt ttataataca acatgactgt ttttaaagta
     1081 caaaataagt ttatttttgt aaaagagaga atgtttaaaa gttttgttac tttatagaag
     1141 aaattttgag tttttgtttt tttttaataa ataaataaac ataaataaat tgtttgttga
     1201 atttattatt agtatgtaag tgtaaatata ataaaactta atatctattc aaattaataa
     1261 ataaacctcg atatacagac cgataaaaca catgcgtcaa ttttacgcat gattatcttt
     1321 aacgtacgtc acaatatgat tatctttcta gggttaaata atagtttcta atttttttat
     1381 tattcagcct gctgtcgtga ataccgagct ccaattcgcc ctatagtgag tcgtattaca
     1441 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     1501 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     1561 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     1621 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     1681 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     1741 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     1801 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     1861 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     1921 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     1981 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     2041 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2101 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2161 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     2221 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     2281 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     2341 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     2401 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     2461 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     2521 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     2581 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     2641 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     2701 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     2761 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     2821 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     2881 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     2941 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3001 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     3061 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3121 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3181 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3241 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3301 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3361 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3421 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3481 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3541 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3601 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3661 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3721 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3781 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3841 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3901 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3961 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     4021 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     4081 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     4141 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     4201 cagctatgac catgattacg ccaagctcga aattaaccct cactaaaggg aacaaaagct
     4261 ggtacctcgc gcgacttggt ttgccattct ttagcgcgcg tcgcgtcaca cagcttggcc
     4321 acaatgtggt ttttgtcaaa cgaagattct atgacgtgtt taaagtttag gtcgagtaaa
     4381 gcgcaaatct tttttaaccc tagaaagata gtctgcgtaa aattgacgca tgcattcttg
     4441 aaatattgct ctctctttct aaatagcgcg aatccgtcgc tgtgcattta ggacatctca
     4501 gtcgccgctt ggagctcccg tgaggcgtgc ttgtcaatgc ggtaagtgtc actgattttg
     4561 aactataacg accgcgtgag tcaaaatgac gcatgattat cttttacgtg acttttaaga
     4621 tttaactcat acgataatta tattgttatt tcatgttcta cttacgtgat aacttattat
     4681 atatatattt tcttgttata gatatcgtga ctaatatata ataaaatggg tagttcttta
     4741 gacgatgagc atatcctctc tgctcttctg caaagcgatg acgagcttgt tggctagcgg
     4801 cgcgccgtcg agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     4861 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     4921 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     4981 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt
     5041 atgatctcta gtcaaggcac tatacatcaa atattcctta ttaacccctt tacaaattaa
     5101 aaagctaaag gtacacaatt tttgagcata gttattaata gcagacactc tatgcctgtg
     5161 tggagtaaga aaaaacagta tgttatgatt ataactgtta tgcctactta taaaggttac
     5221 agaatatttt tccataattt tcttgtatag cagtgcagct ttttctctag agctcaggca
     5281 ccgggcttgc gggtcatgca ccaggtgcgc ggtccttcgg gcacctcgac gtcggcggtg
     5341 acggtgaagc cgagccgctc gtagaagggg aggttgcggg gcgcggaggt ctccaggaag
     5401 gcgggcaccc cggcgcgctc ggccgcctcc actccgggga gcacgacggc gctgcccaga
     5461 cccttgccct ggtggtcggg cgagacgccg acggtggcca ggaaccacgc gggctccttg
     5521 ggccggtgcg gcgccaggag gccttccatc tgttgctgcg cggccagccg ggaaccgctc
     5581 aactcggcca tgcgcgggcc gatctcggcg aacaccgccc ccgcttcgac gctctccggc
     5641 gtggtccaga ccgccaccgc ggcgccgtcg tccgcgaccc acaccttgcc gatgtcgagc
     5701 ccgacgcgcg tgaggaagag ttcttgcagc tcggtgaccc gctcgatgtg gcggtccggg
     5761 tcgacggtgt ggcgcgtggc ggggtagtcg gcgaacgcgg cggcgagggt gcgtacggcc
     5821 cgggggacgt cgtcgcgggt ggcgaggcgc accgtgggct tgtactcggt
//
