LOCUS       sequence-343598-        4345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for making Rluc mRNA reporter with pA120..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4345)
  AUTHORS   Xiang K, Bartel DP
  TITLE     The molecular basis of coupling between poly(A)-tail length and 
            translational efficiency.
  JOURNAL   Elife. 2021 Jul 2;10. pii: 66493. doi: 10.7554/eLife.66493.
  PUBMED    34213414
REFERENCE   2  (bases 1 to 4345)
  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: "Elife."; 
            date: "2021-07-2"; volume: "10. pii: 66493. doi"; pages: " 
            10.7554/eLife.66493"
COMMENT     Sequence Label: C035
FEATURES             Location/Qualifiers
     source          1..4345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             23..958
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     promoter        complement(1216..1234)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1217..1234)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1252..1268)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1252..1268)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1265..1287)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1276..1292
                     /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)."
     promoter        complement(join(1300..1306,1307..1324,1325..1330))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1345..1366
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1483..1500)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1654..2242)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1734..1753)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2413..3204,3205..3273))
                     /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     3036..3055
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3274..3378)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3446..3464
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3502..3524)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3624..3643
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3710..4165)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3847..3868)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4059..4078
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4291..4313
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4305..4322
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4306..4322
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4329..4345,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(4329..4345,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg gccaagtcgg ccatgacttc gaaagtttat gatccagaac aaaggaaacg
       61 gatgataact ggtccgcagt ggtgggccag atgtaaacaa atgaatgttc ttgattcatt
      121 tattaattat tatgattcag aaaaacatgc agaaaatgct gttatttttt tacatggtaa
      181 cgcggcctct tcttatttat ggcgacatgt tgtgccacat attgagccag tagcgcggtg
      241 tattatacca gaccttattg gtatgggcaa atcaggcaaa tctggtaatg gttcttatag
      301 gttacttgat cattacaaat atcttactgc atggtttgaa cttcttaatt taccaaagaa
      361 gatcattttt gtcggccatg attggggtgc ttgtttggca tttcattata gctatgagca
      421 tcaagataag atcaaagcaa tagttcacgc tgaaagtgta gtagatgtga ttgaatcatg
      481 ggatgaatgg cctgatattg aagaagatat tgcgttgatc aaatctgaag aaggagaaaa
      541 aatggttttg gagaataact tcttcgtgga aaccatgttg ccatcaaaaa tcatgagaaa
      601 gttagaacca gaagaatttg cagcatatct tgaaccattc aaagagaaag gtgaagttcg
      661 tcgtccaaca ttatcatggc ctcgtgaaat cccgttagta aaaggtggta aacctgacgt
      721 tgtacaaatt gttaggaatt ataatgctta tctacgtgca agtgatgatt taccaaaaat
      781 gtttattgaa tcggacccag gattcttttc caatgctatt gttgaaggtg ccaagaagtt
      841 tcctaatact gaatttgtca aagtaaaagg tcttcatttt tcgcaagaag atgcacctga
      901 tgaaatggga aaatatatca aatcgttcgt tgagcgagtt ctcaaaaatg aacaataacg
      961 aattcgtcga cctcgaggga tccgggcgta atccagacat gaaaaaaaaa aaaaaaaaaa
     1021 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     1081 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa agctagccat ggtcccagcc
     1141 tcctcgctgg cggctagtgg gcaacatgct tcggcatggc gaatgggacg cggccgcatg
     1201 cataagcttg agtattctat agtgtcacct aaatagcttg gcgtaatcat ggtcatagct
     1261 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     1321 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     1381 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     1441 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     1501 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     1561 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     1621 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     1681 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     1741 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     1801 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     1861 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     1921 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     1981 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2041 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2101 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2161 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2221 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2281 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2341 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2401 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2461 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     2521 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     2581 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     2641 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     2701 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     2761 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     2821 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     2881 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     2941 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3001 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3061 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3121 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3181 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3241 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     3301 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3361 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     3421 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     3481 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     3541 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     3601 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     3661 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc
     3721 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3781 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3841 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3901 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3961 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4021 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4081 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4141 cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac tgttgggaag
     4201 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     4261 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     4321 gtgaattgta atacgactca ctata
//
