LOCUS       sequence-343184-        4216 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for making Rluc mRNA reporter with no pA..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4216)
  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 4216)
  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: C022
FEATURES             Location/Qualifiers
     source          1..4216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             40..975
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     promoter        complement(1104..1122)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1105..1122)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1140..1156)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1140..1156)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1153..1175)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1164..1180
                     /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(1188..1194,1195..1212,1213..1218))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1233..1254
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1371..1388)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1542..2130)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1622..1641)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2301..3092,3093..3161))
                     /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     2924..2943
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3162..3266)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3334..3352
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3390..3412)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3512..3531
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3598..4053)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3735..3756)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3947..3966
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4179..4201
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4193..4210
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4194..4210
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattggcc aagtcggcca tgacttcgaa agtttatgat
       61 ccagaacaaa ggaaacggat gataactggt ccgcagtggt gggccagatg taaacaaatg
      121 aatgttcttg attcatttat taattattat gattcagaaa aacatgcaga aaatgctgtt
      181 atttttttac atggtaacgc ggcctcttct tatttatggc gacatgttgt gccacatatt
      241 gagccagtag cgcggtgtat tataccagac cttattggta tgggcaaatc aggcaaatct
      301 ggtaatggtt cttataggtt acttgatcat tacaaatatc ttactgcatg gtttgaactt
      361 cttaatttac caaagaagat catttttgtc ggccatgatt ggggtgcttg tttggcattt
      421 cattatagct atgagcatca agataagatc aaagcaatag ttcacgctga aagtgtagta
      481 gatgtgattg aatcatggga tgaatggcct gatattgaag aagatattgc gttgatcaaa
      541 tctgaagaag gagaaaaaat ggttttggag aataacttct tcgtggaaac catgttgcca
      601 tcaaaaatca tgagaaagtt agaaccagaa gaatttgcag catatcttga accattcaaa
      661 gagaaaggtg aagttcgtcg tccaacatta tcatggcctc gtgaaatccc gttagtaaaa
      721 ggtggtaaac ctgacgttgt acaaattgtt aggaattata atgcttatct acgtgcaagt
      781 gatgatttac caaaaatgtt tattgaatcg gacccaggat tcttttccaa tgctattgtt
      841 gaaggtgcca agaagtttcc taatactgaa tttgtcaaag taaaaggtct tcatttttcg
      901 caagaagatg cacctgatga aatgggaaaa tatatcaaat cgttcgttga gcgagttctc
      961 aaaaatgaac aataacgaat tcgtcgacct cgagggatcc gggccctctg ctagccatgg
     1021 tcccagcctc ctcgctggcg gctagtgggc aacatgcttc ggcatggcga atgggacgcg
     1081 gccgcatgca taagcttgag tattctatag tgtcacctaa atagcttggc gtaatcatgg
     1141 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     1201 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     1261 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     1321 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     1381 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1441 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1501 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1561 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1621 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1681 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1741 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1801 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1861 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1921 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1981 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2041 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2101 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2161 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2221 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2281 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2341 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2401 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     2461 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2521 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2581 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     2641 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     2701 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     2761 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     2821 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     2881 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     2941 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3001 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3061 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3121 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3181 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3241 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     3301 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     3361 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     3421 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     3481 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     3541 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa
     3601 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     3661 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     3721 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     3781 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     3841 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     3901 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     3961 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     4021 ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc tgcgcaactg
     4081 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     4141 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     4201 gacggccagt gaattg
//
