LOCUS       sequence-474909-        5756 bp    DNA     circular SYN 06-JUN-2025
DEFINITION  A plasmid IVT template containing a Firefly 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 5756)
  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 5756)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 6, 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_luc-60A
FEATURES             Location/Qualifiers
     source          1..5756
                     /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    1166..1300
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1203..1222)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1257..1276
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1338..2990)
                     /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(3032..3050)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3033..3050)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(3085..4069)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     rep_origin      4140..4595
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4227..4246)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4437..4458
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4621..4725
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4726..4794,4795..5586)
                     /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(4944..4963)
                     /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 cctcccacac ctccccctga acctgaaaca
     1141 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     1201 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     1261 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc tacgtaatac gactcactat
     1321 agttctagag gctcgagtta cacggcgatc ttgccgccct tcttggcctt aatgagaatc
     1381 tcgcggatct tgcgggcgtc caacttgccg gtcagtcctt taggcacctc gtccacgaac
     1441 acaacaccac cgcgcagctt cttggcggtt gtaacctggc tggccacata gtccacgatc
     1501 tccttctcgg tcatggtttt accgtgttcc agcacgacga ctgcggcggg cagctcgccg
     1561 gcatcgtcgt cgggcaggcc ggcgaccccg gcgtcgaaga tgttggggtg ttgcagcagg
     1621 atgctctcca gttcggctgg ggctacctgg tagcccttgt atttgatcag gctcttcagc
     1681 cggtccacga tgaagaagtg ctcgtcctcg tcccagtagg cgatgtcgcc gctgtgcagc
     1741 cagccgtcct tgtcgatgag agcgtttgta gcctcggggt tgttaacgta gccgctcatg
     1801 atcatggggc cacggacgca cagctcgccg cgctggttca cacccagtgt cttaccggtg
     1861 tccaagtcca ccaccttagc ctcgaagaag ggcaccacct tgcctactgc gccaggcttg
     1921 tcgtcccctt cgggggtgat cagaatggcg ctggttgttt ctgtcaggcc gtagccctgg
     1981 cggatgcctg gtaggtggaa gcgtttggcc acggcctcac ctacctcctt gctgagcggc
     2041 gccccgccgc tggcgatctc gtgcaagttg cttaggtcgt acttgtcgat gagagtgctc
     2101 ttagcgaaga agctaaatag tgtgggcacc agcagggcag attgaatctt atagtcttgc
     2161 aagctgcgca agaatagctc ctcctcgaag cggtacatga gcacgacccg aaagccgcag
     2221 atcaagtagc ccagcgtggt gaacatgccg aagccgtggt gaaatggcac cacgctgagg
     2281 atagcggtgt cggggatgat ctggttgccg aagatggggt cgcgggcatg actgaatcgg
     2341 acacaagcgg tgcggtgcgg tagggctacg cccttgggca atccggtact gccactactg
     2401 ttcatgatca gggcgatggt tttgtcccgg tcgaagctct cgggcacgaa gtcgtactcg
     2461 ttgaagccgg gtggcaaatg ggaagtcacg aaggtgtaca tgctttggaa gccctggtag
     2521 tcggtcttgc tatccatgat gatgatcttt tgtatgatcg gtagcttctt ttgcacgttg
     2581 aggatctttt gcagcccttt cttgctcacg aatacgacgg tgggctggct gatgcccatg
     2641 ctgttcagca gctcgcgctc gttgtagatg tcgttagctg gggccacagc cacaccgatg
     2701 aacagggcac ccaacacggg catgaagaac tgcaagctat tctcgctgca caccacgatc
     2761 cgatggtttg tattcagccc atagcgcttc atagcttctg ccagccgaac gctcatctcg
     2821 aagtactcgg cgtaggtaat gtccacctcg atatgtgcgt cggtaaaggc gatggtgccg
     2881 ggcaccaggg cgtagcgctt catggctttg tgcagctgct cgccggcggt cccgtcttcg
     2941 agtgggtaga atggcgctgg gcccttctta atgtttttgg catcttccat cgaatcgatg
     3001 ggatcctgca aaaagaacaa gtagcttgta ttctatagtg tcacctaaat caagcttgct
     3061 ccgacgtccc caggcagaat ggcggttccc taaacgagca ttgcttatat agacctccca
     3121 ttaggcacgc ctaccgccca tttacgtcaa tggaacgccc atttgcgtca ttgcccctcc
     3181 ccattgacgt caatggggat gtacttggca gccatcgcgg gccatttacc gccattgacg
     3241 tcaatgggag tactgccaat gtaccctggc gtacttccaa tagtaatgta cttgccaagt
     3301 tactattaat agatattgat gtactgccaa gtgggccatt taccgtcatt gacgtcaata
     3361 gggggcgtga gaacggatat gaatgggcaa tgagccatcc cattgacgtc aatggtgggt
     3421 ggtcctattg acgtcaatgg gcattgagcc aggcgggcca tttaccgtaa ttgacgtcaa
     3481 tgggggaggc gccatatacg tcaataggac cgcccatatg acgtcaatag gaaagaccat
     3541 atatagagac cattgacgtc aatgggggag tggctatggg cggtattagg aagccccata
     3601 tatggtatat gggaccgccc attgggaggg gctatctacg tcaataggaa aacccatata
     3661 tggaatacta tatggcatag ggccaataca tagtattgaa cctggccaat agccatattg
     3721 gcatagggcc atattggata ttgcctatat attgatcctg gcatatagcc aatatggccg
     3781 ccattattgg caccatgcca attcaatatg gcggacttgg cacagggcca agacccctcc
     3841 tcaaacttgg caccgtgcca agtagacccc tccccagttg gcacagtgcc aggtccgcca
     3901 tattgaattg gcatggtgcc aagtccgcca tattgaattg gctggggtcc agatccacca
     3961 tattgaattg gcacgagtcc atatacgcca tattgaattg gcacaggtcc agatccacca
     4021 tattgaattg gcatgagtcc atatacgcca tattgaattg gcggtcgact ggcgtaatag
     4081 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     4141 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4201 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4261 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4321 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4381 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4441 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4501 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4561 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     4621 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4681 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4741 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4801 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4861 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4921 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4981 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5041 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5101 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5161 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5221 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5281 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5341 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5401 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5461 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5521 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5581 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5641 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5701 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttc
//
