LOCUS       sequence-343601-        4256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for making Rluc mRNA reporter with pA30..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4256)
  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 4256)
  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: C031
FEATURES             Location/Qualifiers
     source          1..4256
                     /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(1127..1145)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1128..1145)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1163..1179)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1163..1179)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1176..1198)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1187..1203
                     /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(1211..1217,1218..1235,1236..1241))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1256..1277
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1394..1411)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1565..2153)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1645..1664)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2324..3115,3116..3184))
                     /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     2947..2966
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3185..3289)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3357..3375
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3413..3435)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3535..3554
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3621..4076)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3758..3779)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3970..3989
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4202..4224
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4216..4233
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4217..4233
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4240..4256,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(4240..4256,1..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 aagctagcca tggtcccagc ctcctcgctg gcggctagtg ggcaacatgc
     1081 ttcggcatgg cgaatgggac gcggccgcat gcataagctt gagtattcta tagtgtcacc
     1141 taaatagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     1201 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     1261 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     1321 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     1381 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     1441 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     1501 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1561 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1621 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1681 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1741 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1801 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1861 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1921 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1981 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2041 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2101 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2161 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2221 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2281 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     2341 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     2401 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     2461 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     2521 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     2581 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     2641 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     2701 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     2761 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     2821 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     2881 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     2941 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3001 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     3061 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     3121 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     3181 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     3241 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     3301 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     3361 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     3421 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     3481 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     3541 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     3601 ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     3661 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     3721 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     3781 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     3841 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     3901 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     3961 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     4021 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat
     4081 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     4141 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     4201 tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc actata
//
