LOCUS       sequence-343596-        4982 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for making mRNA for overexpressing frog RPL3 (C-HA) in frog
            oocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4982)
  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 4982)
  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: C245
FEATURES             Location/Qualifiers
     source          1..4982
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(348..366)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(349..366)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(384..400)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(384..400)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(397..419)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    408..424
                     /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(432..438,439..456,457..462))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    477..498
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(615..632)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(786..1374)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(866..885)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1545..2336,2337..2405))
                     /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     2168..2187
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2406..2510)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2578..2596
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2634..2656)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2756..2775
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2842..3297)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2979..3000)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3191..3210
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3423..3445
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3437..3454
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3438..3454
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3461..3480
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3461..3479
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           3493..3535
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             4756..4782
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     3'UTR           4785..4910
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
ORIGIN
        1 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
       61 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
      121 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aacagtaggg ttgtaaaggt ttttcttttc
      181 ctgagaaaac aaccttttgt tttctcaggt tttgcttttt ggcctttccc tagctttaaa
      241 aaaaaaagca aaagctagcc atggtcccag cctcctcgct ggcggctagt gggcaacatg
      301 cttcggcatg gcgaatggga cgcggccgca tgcataagct tgagtattct atagtgtcac
      361 ctaaatagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
      421 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
      481 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
      541 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
      601 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
      661 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
      721 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
      781 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
      841 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
      901 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
      961 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     1021 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     1081 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     1141 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     1201 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     1261 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     1321 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     1381 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     1441 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     1501 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     1561 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     1621 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     1681 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     1741 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     1801 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     1861 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     1921 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     1981 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     2041 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     2101 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     2161 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     2221 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     2281 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     2341 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     2401 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     2461 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     2521 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     2581 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     2641 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     2701 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     2761 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     2821 aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2881 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2941 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     3001 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     3061 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     3121 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     3181 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3241 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca
     3301 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     3361 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     3421 ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact cactataggg
     3481 gaatacaagc ttcttgttct ttttgcagaa gctcagaata aacgctcaac tttggcagat
     3541 ctgaacatgt ctcaccgaaa gttctcagcc ccccgacatg gatccctggg cttcttgccc
     3601 cgcaaacgtt gtaagagaca tcgtggcaaa gtgaagagct tccccaagga tgacccaagc
     3661 aaacccattc acctgaccgc tttcttgggc tacaaggctg gcatgactca cattgttcgt
     3721 gaggtcgacc gacctggctc aaaggtgaac aagaaagagg ttgtggaagc agtaactgtg
     3781 gttgaaacac ctcccatggt aattgttggt attgttggct atgtccagac tcctcgtggt
     3841 ctgcgcagct tgaagaccat ctttgctgag cacatcagtg atgaatgcaa gaggcgcttc
     3901 tacaagaact ggtacaaatc caagaagaag gcgttcacaa agtactgcaa gaaatggcag
     3961 gatgaagacg gaaagaaaca gctggagaag gactttgcta gcatgaaaaa gtactgccaa
     4021 gtcatcaggg ttattgccca tacacagatg cgtctgcttc ccctgcgcca gaagaagtct
     4081 cacctcatgg agattcaagt gaacggaggt acaattgcag agaaggtgga ctgggcccgt
     4141 gagaaactgg agcaacaagt tgtaattgga ggagtgttcg ggcaggatga aatgattgat
     4201 gtgattggtg tcactaaagg aaaaggctac aaaggtgtaa ctagccgttg gcacacaaag
     4261 aaactgcccc gtaagactca cagaggtctt cgtaaagtgg cttgtattgg agcttggcat
     4321 cctgcccgtg tggctttctc agtagcccgt gctggacaga agggatacca ccaccgtaca
     4381 gagattaaca aaaagattta caaaattggc cagggctacc attcaaagga tggtaaactt
     4441 gtgaagaaca atgcttcaac agactacgat ctatctgaca agagcatcaa ccctctgggt
     4501 gggtttgttc attatggaga ggtgaaaaat gactttatca tgctgaaagg ctgtgttatt
     4561 gggaccaaga agagagtact cacactgcgc aagtccctcc ttgtgcaaac cagtcgccgt
     4621 gccctggaga agattgatct caagtttatt gatacaacat ccaagtttgg gcatggtcgt
     4681 ttccagacag tagaggaaaa gaaggctttc atgggaccac tcaagaaaga ccgtttggca
     4741 aaagaggaaa ctgcatatcc atatgatgtt ccagattatg cttaaaccag cctcaagaac
     4801 acccgaatgg agtctctaag ctacataata ccaacttaca ctttacaaaa tgttgtcccc
     4861 caaaatgtag ccattcgtat ctgctcctaa taaaaagaaa gtttcttcac attctgggcg
     4921 tagatccaga catgaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     4981 aa
//
