LOCUS       sequence-261145-        4199 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Csy4 in Mammalian cells for site-specific RNA processing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4199)
  AUTHORS   Nissim L, Perli SD, Fridkin A, Perez-Pinera P, Lu TK
  TITLE     Multiplexed and Programmable Regulation of Gene Networks with an 
            Integrated RNA and CRISPR/Cas Toolkit in Human Cells.
  JOURNAL   Mol Cell. 2014 May 14. pii: S1097-2765(14)00355-4. doi: 
            10.1016/j.molcel.2014.04.022.
  PUBMED    24837679
REFERENCE   2  (bases 1 to 4199)
  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: "Mol Cell. 
            2014 May 14. pii: S1097-2765(14)00355-4. doi: 
            10.1016/j.molcel.2014.04.022."
COMMENT     Sequence Label: PGK1p-Csy4-pA (Construct 2)
FEATURES             Location/Qualifiers
     source          1..4199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(88..107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        589..1088
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(624..647)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1021..1040
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             1130..1147
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1148..1168
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     polyA_signal    1914..2035
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1925..1944)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1979..1998
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2278..2295)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2449..3037)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2529..2548)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3208..3999,4000..4068))
                     /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     3831..3850
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4069..4173)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttgcca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagccc aagctaccat
      541 gataagtaag taatattaag gtacgggagg tacttggagg tacctaccgg gtaggggagg
      601 cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc acttggcgct
      661 acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc caaccggctc
      721 cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc
      781 ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt ctcactagtc
      841 tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc
      901 aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt gggtccgggg
      961 gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg
     1021 cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg
     1081 gcctttcgac ctgcatccca agcttacgcg tgccaccatg aaatcttctc accatcacca
     1141 tcaccatgaa aacctgtact tccaatccaa tgcagctagc gaccactatc tggacatcag
     1201 actgaggccc gatcctgagt tccctcccgc ccagctgatg agcgtgctgt ttggcaagct
     1261 gcatcaggct ctggtcgccc aaggcggaga cagaatcggc gtgtccttcc ccgacctgga
     1321 cgagtcccgg agtcgcctgg gcgagcggct gagaatccac gccagcgcag acgatctgcg
     1381 cgccctgctg gcccggcctt ggctggaggg cctgcgggat catctgcagt ttggcgagcc
     1441 cgccgtggtg ccacacccaa caccctaccg ccaggtgagc cgcgtgcagg ccaagtcaaa
     1501 tcccgagaga ctgcggcgga ggctgatgag gcgacatgat ctgagcgagg aggaggccag
     1561 aaagagaatc cccgacacag tggccagagc cctggatctg ccatttgtga ccctgcggag
     1621 ccagagcact ggccagcatt tcagactgtt catcagacac gggcccctgc aggtgacagc
     1681 cgaggagggc ggatttacat gctatggcct gtctaaaggc ggcttcgtgc cctggttctg
     1741 atgagagctc taagcggccg cataagatat cagccatggc ttcccgccgg cggtggcggc
     1801 gcaggatgat ggcacgctgc ccatgtcttg tgcccaggag agcgggatgg accgtcaccc
     1861 tgcagcctgt gcttctgcta ggatcaatgt gtaggctagt cgagcagaca tgataagata
     1921 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     1981 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     2041 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     2101 caagtaaaac ctctacaaat gtggtaaaat cgagtatgtc gaccgatgcc cttgagagcc
     2161 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     2221 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc
     2281 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2341 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2401 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2461 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2521 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2581 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2641 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2701 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2761 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2821 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2881 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2941 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3001 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3061 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3121 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3181 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3241 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3301 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3361 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3421 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3481 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3541 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3601 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3661 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3721 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3781 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3841 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     3901 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     3961 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4021 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4081 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4141 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctg
//
