LOCUS       sequence-402534-        5905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A plasmid IVT template containing a destabilized Firefly 
            luciferase-PEST (degron) fusion followed by a template-encoded 
            poly(A) tail. Linearization at an Esp3i site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5905)
  AUTHORS   Aditham A, Shi H, Guo J, Zeng H, Zhou Y, Wade SD, Huang J, Liu J, 
            Wang X
  TITLE     Chemically Modified mocRNAs for Highly Efficient Protein Expression 
            in Mammalian Cells.
  JOURNAL   ACS Chem Biol. 2022 Dec 16;17(12):3352-3366. doi: 
            10.1021/acschembio.1c00569. Epub 2022 Jan 7.
  PUBMED    34995053
REFERENCE   2  (bases 1 to 5905)
  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: 
            "10.1021/acschembio.1c00569"; journalName: "ACS Chem Biol"; date: 
            "2022-12-16- 16"; volume: "17"; issue: "12"; pages: "3352-3366"
COMMENT     Sequence Label: pCS2_Firefly_degron-60A
FEATURES             Location/Qualifiers
     source          1..5905
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    1195..1329
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1232..1251)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1286..1305
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1370..1489)
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     CDS             complement(1490..3139)
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     promoter        complement(3181..3199)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3182..3199)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(3234..4218)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     rep_origin      4289..4744
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4376..4395)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4586..4607
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4770..4874
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4875..4943,4944..5735)
                     /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(5093..5112)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgctccc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tgggtaccgg gcccaatgca ttggcgccgc ggccgcgaac
     1081 gtctcttttt tttttttttt tttttttttt tttttttttt tttttttttt tttttttttc
     1141 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     1201 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     1261 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     1321 gtctggatct acgtaatacg actcactata gttctagagg ctcgagttag acgttgatgc
     1381 gagctgaagc acaagcggct gggtgccggt ccataccgct ttcttgtgcg caggacatag
     1441 gcaatgtacc ggcggcttgc tcttcaacct caggcggaaa gccgtgagac acggcgatct
     1501 tgccgccctt cttggcctta atgagaatct cgcggatctt gcgggcgtcc aacttgccgg
     1561 tcagtccttt aggcacctcg tccacgaaca caacaccacc gcgcagcttc ttggcggttg
     1621 taacctggct ggccacatag tccacgatct ccttctcggt catggtttta ccgtgttcca
     1681 gcacgacgac tgcggcgggc agctcgccgg catcgtcgtc gggcaggccg gcgaccccgg
     1741 cgtcgaagat gttggggtgt tgcagcagga tgctctccag ttcggctggg gctacctggt
     1801 agcccttgta tttgatcagg ctcttcagcc ggtccacgat gaagaagtgc tcgtcctcgt
     1861 cccagtaggc gatgtcgccg ctgtgcagcc agccgtcctt gtcgatgaga gcgtttgtag
     1921 cctcggggtt gttaacgtag ccgctcatga tcatggggcc acggacgcac agctcgccgc
     1981 gctggttcac acccagtgtc ttaccggtgt ccaagtccac caccttagcc tcgaagaagg
     2041 gcaccacctt gcctactgcg ccaggcttgt cgtccccttc gggggtgatc agaatggcgc
     2101 tggttgtttc tgtcaggccg tagccctggc ggatgcctgg taggtggaag cgtttggcca
     2161 cggcctcacc tacctccttg ctgagcggcg ccccgccgct ggcgatctcg tgcaagttgc
     2221 ttaggtcgta cttgtcgatg agagtgctct tagcgaagaa gctaaatagt gtgggcacca
     2281 gcagggcaga ttgaatctta tagtcttgca agctgcgcaa gaatagctcc tcctcgaagc
     2341 ggtacatgag cacgacccga aagccgcaga tcaagtagcc cagcgtggtg aacatgccga
     2401 agccgtggtg aaatggcacc acgctgagga tagcggtgtc ggggatgatc tggttgccga
     2461 agatggggtc gcgggcatga ctgaatcgga cacaagcggt gcggtgcggt agggctacgc
     2521 ccttgggcaa tccggtactg ccactactgt tcatgatcag ggcgatggtt ttgtcccggt
     2581 cgaagctctc gggcacgaag tcgtactcgt tgaagccggg tggcaaatgg gaagtcacga
     2641 aggtgtacat gctttggaag ccctggtagt cggtcttgct atccatgatg atgatctttt
     2701 gtatgatcgg tagcttcttt tgcacgttga ggatcttttg cagccctttc ttgctcacga
     2761 atacgacggt gggctggctg atgcccatgc tgttcagcag ctcgcgctcg ttgtagatgt
     2821 cgttagctgg ggccacagcc acaccgatga acagggcacc caacacgggc atgaagaact
     2881 gcaagctatt ctcgctgcac accacgatcc gatggtttgt attcagccca tagcgcttca
     2941 tagcttctgc cagccgaacg ctcatctcga agtactcggc gtaggtaatg tccacctcga
     3001 tatgtgcgtc ggtaaaggcg atggtgccgg gcaccagggc gtagcgcttc atggctttgt
     3061 gcagctgctc gccggcggtc ccgtcttcga gtgggtagaa tggcgctggg cccttcttaa
     3121 tgtttttggc atcttccatc gaatcgatgg gatcctgcaa aaagaacaag tagcttgtat
     3181 tctatagtgt cacctaaatc aagcttgctc cgacgtcccc aggcagaatg gcggttccct
     3241 aaacgagcat tgcttatata gacctcccat taggcacgcc taccgcccat ttacgtcaat
     3301 ggaacgccca tttgcgtcat tgcccctccc cattgacgtc aatggggatg tacttggcag
     3361 ccatcgcggg ccatttaccg ccattgacgt caatgggagt actgccaatg taccctggcg
     3421 tacttccaat agtaatgtac ttgccaagtt actattaata gatattgatg tactgccaag
     3481 tgggccattt accgtcattg acgtcaatag ggggcgtgag aacggatatg aatgggcaat
     3541 gagccatccc attgacgtca atggtgggtg gtcctattga cgtcaatggg cattgagcca
     3601 ggcgggccat ttaccgtaat tgacgtcaat gggggaggcg ccatatacgt caataggacc
     3661 gcccatatga cgtcaatagg aaagaccata tatagagacc attgacgtca atgggggagt
     3721 ggctatgggc ggtattagga agccccatat atggtatatg ggaccgccca ttgggagggg
     3781 ctatctacgt caataggaaa acccatatat ggaatactat atggcatagg gccaatacat
     3841 agtattgaac ctggccaata gccatattgg catagggcca tattggatat tgcctatata
     3901 ttgatcctgg catatagcca atatggccgc cattattggc accatgccaa ttcaatatgg
     3961 cggacttggc acagggccaa gacccctcct caaacttggc accgtgccaa gtagacccct
     4021 ccccagttgg cacagtgcca ggtccgccat attgaattgg catggtgcca agtccgccat
     4081 attgaattgg ctggggtcca gatccaccat attgaattgg cacgagtcca tatacgccat
     4141 attgaattgg cacaggtcca gatccaccat attgaattgg catgagtcca tatacgccat
     4201 attgaattgg cggtcgactg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     4261 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     4321 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4381 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4441 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4501 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4561 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4621 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4681 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     4741 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4801 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4861 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4921 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4981 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5041 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5101 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5161 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5221 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5281 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5341 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5401 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5461 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5521 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5581 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5641 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5701 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5761 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5821 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5881 ccgtagaaaa gatcaaagga tcttc
//
