LOCUS       sequence-343599-        4289 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for making Rluc mRNA reporter with pA63..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4289)
  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 4289)
  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: C033
FEATURES             Location/Qualifiers
     source          1..4289
                     /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(1160..1178)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1161..1178)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1196..1212)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1196..1212)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1209..1231)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1220..1236
                     /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(1244..1250,1251..1268,1269..1274))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1289..1310
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1427..1444)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1598..2186)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1678..1697)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2357..3148,3149..3217))
                     /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     2980..2999
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3218..3322)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3390..3408
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3446..3468)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3568..3587
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3654..4109)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3791..3812)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4003..4022
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4235..4257
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4249..4266
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4250..4266
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4273..4289,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(4273..4289,1..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 gatccagaca tgaaaaaaaa aaaaaaaaaa
     1021 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaagctag ccatggtccc
     1081 agcctcctcg ctggcggcta gtgggcaaca tgcttcggca tggcgaatgg gacgcggccg
     1141 catgcataag cttgagtatt ctatagtgtc acctaaatag cttggcgtaa tcatggtcat
     1201 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     1261 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     1321 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     1381 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     1441 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     1501 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     1561 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     1621 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     1681 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1741 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     1801 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1861 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1921 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1981 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     2041 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     2101 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     2161 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2221 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     2281 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     2341 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     2401 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     2461 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     2521 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     2581 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     2641 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     2701 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     2761 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     2821 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     2881 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     2941 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     3001 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     3061 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     3121 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3181 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     3241 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     3301 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     3361 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     3421 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     3481 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     3541 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     3601 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt
     3661 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     3721 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3781 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3841 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3901 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3961 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     4021 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     4081 cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg
     4141 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     4201 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     4261 gccagtgaat tgtaatacga ctcactata
//
