LOCUS       sequence-343300-        4446 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for making Rluc mRNA reporter with pA98..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4446)
  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 4446)
  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: C034
FEATURES             Location/Qualifiers
     source          1..4446
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             12..947
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     promoter        complement(1306..1324)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1307..1324)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1342..1358)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1342..1358)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1355..1377)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1366..1382
                     /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(1390..1396,1397..1414,1415..1420))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1435..1456
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1573..1590)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1744..2332)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1824..1843)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2503..3294,3295..3363))
                     /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     3126..3145
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3364..3468)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3536..3554
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3592..3614)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3714..3733
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3800..4255)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3937..3958)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4149..4168
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4381..4403
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4395..4412
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4396..4412
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4419..4438
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4419..4437
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ccaagtcggc catgacttcg aaagtttatg atccagaaca aaggaaacgg atgataactg
       61 gtccgcagtg gtgggccaga tgtaaacaaa tgaatgttct tgattcattt attaattatt
      121 atgattcaga aaaacatgca gaaaatgctg ttattttttt acatggtaac gcggcctctt
      181 cttatttatg gcgacatgtt gtgccacata ttgagccagt agcgcggtgt attataccag
      241 accttattgg tatgggcaaa tcaggcaaat ctggtaatgg ttcttatagg ttacttgatc
      301 attacaaata tcttactgca tggtttgaac ttcttaattt accaaagaag atcatttttg
      361 tcggccatga ttggggtgct tgtttggcat ttcattatag ctatgagcat caagataaga
      421 tcaaagcaat agttcacgct gaaagtgtag tagatgtgat tgaatcatgg gatgaatggc
      481 ctgatattga agaagatatt gcgttgatca aatctgaaga aggagaaaaa atggttttgg
      541 agaataactt cttcgtggaa accatgttgc catcaaaaat catgagaaag ttagaaccag
      601 aagaatttgc agcatatctt gaaccattca aagagaaagg tgaagttcgt cgtccaacat
      661 tatcatggcc tcgtgaaatc ccgttagtaa aaggtggtaa acctgacgtt gtacaaattg
      721 ttaggaatta taatgcttat ctacgtgcaa gtgatgattt accaaaaatg tttattgaat
      781 cggacccagg attcttttcc aatgctattg ttgaaggtgc caagaagttt cctaatactg
      841 aatttgtcaa agtaaaaggt cttcattttt cgcaagaaga tgcacctgat gaaatgggaa
      901 aatatatcaa atcgttcgtt gagcgagttc tcaaaaatga acaataacga attcgtcgac
      961 ctcgagggat ccgggcgtag atccagacat gaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     1021 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     1081 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     1141 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     1201 aaaaaaaaaa agctagccat ggtcccagcc tcctcgctgg cggctagtgg gcaacatgct
     1261 tcggcatggc gaatgggacg cggccgcatg cataagcttg agtattctat agtgtcacct
     1321 aaatagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     1381 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     1441 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     1501 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1561 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1621 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1681 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1741 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1801 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1861 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1921 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1981 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2041 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2101 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2161 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2221 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2281 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2341 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2401 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2461 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2521 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2581 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2641 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2701 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2761 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2821 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2881 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2941 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3001 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3061 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3121 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3181 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3241 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3301 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3361 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3421 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3481 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     3541 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     3601 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     3661 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     3721 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     3781 gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     3841 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3901 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     3961 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     4021 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     4081 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4141 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     4201 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt
     4261 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     4321 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     4381 cccagtcacg acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg
     4441 aattgg
//
