LOCUS       sequence-393002-        5074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A plasmid IVT template containing a Renilla luciferase CDS 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 5074)
  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 5074)
  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_Renilla_luc-60A
FEATURES             Location/Qualifiers
     source          1..5074
                     /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    1201..1335
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1238..1257)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1292..1311
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1373..2308)
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     promoter        complement(2350..2368)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2351..2368)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(2403..3387)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     rep_origin      3458..3913
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3545..3564)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3755..3776
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3939..4043
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4044..4112,4113..4904)
                     /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(4262..4281)
                     /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 tttttttttt
     1141 tttttcctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     1201 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     1261 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     1321 tatcatgtct ggatctacgt aatacgactc actatagttc tagaggctcg agttactgct
     1381 cgttcttcag cacgcgctcc acgaagctct tgatgtactt acccatttca tctggagcgt
     1441 cctcctggct gaagtggagg cccttcacct tcacgaactc ggtgttaggg aacttcttag
     1501 ctccctcgac aatagcgttg gaaaagaacc cagggtcgga ctcgatgaac atcttaggca
     1561 gatcgtcgct ggcccgaagg taggcgttgt agttgcggac aatctggacg acgtcgggct
     1621 tgcctccctt aacgagaggg atctcgcgag gccaggagag ggtaggccgt ctaacctcgc
     1681 ccttctcctt gaatggctcc aggtaggcag cgaactcctc aggctccagt ttccgcatga
     1741 tcttgcttgg gagcatggtc tcgacgaaga agttattctc aagcaccatt ttctcgccct
     1801 cttcgctctt gatcagggcg atatcctcct cgatgtcagg ccactcgtcc caggactcga
     1861 tcacgtccac gacactctca gcatggacga tggccttgat cttgtcttgg tgctcgtagg
     1921 agtagtgaaa ggccagacaa gccccccagt cgtggcccac aaagatgatt ttctttggaa
     1981 ggttcagcag ctcgaaccaa gcggtgaggt acttgtagtg atccaggagg cgatatgagc
     2041 cattcccgct cttgccggac ttacccattc cgatcagatc agggatgatg catctagcca
     2101 cgggctcgat gtgaggcacg acgtgcctcc acaggtagct ggaggcagcg ttaccatgca
     2161 gaaaaatcac ggcgttctcg gcgtgcttct cggaatcata gtagttgatg aaggagtcca
     2221 gcacgttcat ttgcttgcag cgagcccacc actgaggccc agtgatcatg cgtttgcgtt
     2281 gctcggggtc gtacaccttg gaagccatcg aatcgatggg atcctgcaaa aagaacaagt
     2341 agcttgtatt ctatagtgtc acctaaatca agcttgctcc gacgtcccca ggcagaatgg
     2401 cggttcccta aacgagcatt gcttatatag acctcccatt aggcacgcct accgcccatt
     2461 tacgtcaatg gaacgcccat ttgcgtcatt gcccctcccc attgacgtca atggggatgt
     2521 acttggcagc catcgcgggc catttaccgc cattgacgtc aatgggagta ctgccaatgt
     2581 accctggcgt acttccaata gtaatgtact tgccaagtta ctattaatag atattgatgt
     2641 actgccaagt gggccattta ccgtcattga cgtcaatagg gggcgtgaga acggatatga
     2701 atgggcaatg agccatccca ttgacgtcaa tggtgggtgg tcctattgac gtcaatgggc
     2761 attgagccag gcgggccatt taccgtaatt gacgtcaatg ggggaggcgc catatacgtc
     2821 aataggaccg cccatatgac gtcaatagga aagaccatat atagagacca ttgacgtcaa
     2881 tgggggagtg gctatgggcg gtattaggaa gccccatata tggtatatgg gaccgcccat
     2941 tgggaggggc tatctacgtc aataggaaaa cccatatatg gaatactata tggcataggg
     3001 ccaatacata gtattgaacc tggccaatag ccatattggc atagggccat attggatatt
     3061 gcctatatat tgatcctggc atatagccaa tatggccgcc attattggca ccatgccaat
     3121 tcaatatggc ggacttggca cagggccaag acccctcctc aaacttggca ccgtgccaag
     3181 tagacccctc cccagttggc acagtgccag gtccgccata ttgaattggc atggtgccaa
     3241 gtccgccata ttgaattggc tggggtccag atccaccata ttgaattggc acgagtccat
     3301 atacgccata ttgaattggc acaggtccag atccaccata ttgaattggc atgagtccat
     3361 atacgccata ttgaattggc ggtcgactgg cgtaatagcg aagaggcccg caccgatcgc
     3421 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     3481 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3541 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3601 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3661 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3721 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3781 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3841 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     3901 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3961 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4021 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4081 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4141 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4201 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4261 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4321 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4381 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4441 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4501 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4561 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4621 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4681 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4741 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4801 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4861 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4921 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4981 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5041 cgtcagaccc cgtagaaaag atcaaaggat cttc
//
