LOCUS       sequence-343316-        5571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for making mRNA for overexpressing frog PABPC1 (C-FLAG) in 
            frog oocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5571)
  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 5571)
  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: C077
FEATURES             Location/Qualifiers
     source          1..5571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     5'UTR           16..58
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             1969..1992
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     3'UTR           1995..2120
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(2442..2460)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2443..2460)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2478..2494)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2478..2494)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2491..2513)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2502..2518
                     /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(2526..2532,2533..2550,2551..2556))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2571..2592
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2709..2726)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2880..3468)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2960..2979)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3639..4430,4431..4499))
                     /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     4262..4281
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4500..4604)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4672..4690
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4728..4750)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4850..4869
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4936..5391)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5073..5094)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5285..5304
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5517..5539
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5531..5548
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5532..5548
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5555..5571,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(5555..5571,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaataca agcttcttgt tctttttgca gaagctcaga ataaacgctc aactttggca
       61 gatctgaaca tgaatcccag tgctcccagc tacccaatgg cttccctgta cgttggagac
      121 cttcatcaag atgtaacgga ggccatgctt tatgagaagt ttagcccagc tggtcccatt
      181 ttgtcaatcc gagtttgcag agatatgatt acacgacgct cacttggata tgcatatgtc
      241 aacttccagc aaccagctga tgctgaacgt gctttggata caatgaactt tgatgtcata
      301 aagggcaggc ccgtgcggat tatgtggtct cagcgtgacc cttctctgcg taagagtgga
      361 gttggtaaca ttttcatcaa aaacctggac aagtctattg ataacaaagc cttgtacgat
      421 acattttctg catttggaaa cattctttcc tgtaaggtgg tttgtgatga aaatggatcc
      481 aagggctatg gttttgttca ctttgagaca caagaggctg ctgagagggc tattgataaa
      541 atgaatggca tgcttctcaa tgaccgcaaa gtatttgttg ggcgctttaa gtcccgcaaa
      601 gagcgtgaag ctgagcttgg tgccagagct aaggaattta caaatgttta catcaaaaat
      661 tttggagacg acatgaatga cgagaggctc aaagaaatgt ttggcaaata tgggccagct
      721 cttagtgtta aagttatgac tgatgacaat ggaaagtcaa aaggctttgg ctttgtcagc
      781 tttgaaagac atgaagatgc acaaaaggct gttgatgaaa tgtatggcaa ggatatgaat
      841 gggaagtcca tgtttgtggg ccgtgcacag aaaaaggtgg aaaggcaaac tgagcttaag
      901 cgcaagtttg aacaaatgaa tcaggaccga atcaccagat accagggtgt taacctctat
      961 gttaaaaacc ttgatgatgg tattgatgat gaacgactgc ggaaagaatt cttacctttt
     1021 ggtacaatca ccagtgctaa ggtaatgatg gaaggtggtc gcagtaaagg ctttggtttt
     1081 gtatgctttt cttcacctga agaggccact aaagcagtca cagaaatgaa tggtagaatt
     1141 gttgccacaa aacccctgta tgttgcattg gcccaaagaa aagaagagcg tcaggctcac
     1201 ttgaccaacc agtacatgca gaggatggca agtgttcgtg taccaaatcc tgtgatcaac
     1261 ccataccagc caccaccatc cagctatttc atggcagcta tcccaccggc tcaaaaccgt
     1321 gctgcgtact acccacctgg gcagattgca cagctcaggc ccagcccccg ttggactgca
     1381 cagggtgcca gacctcatcc attccagaac atgcccggag caatccgccc tactgctcca
     1441 agaccaccaa cattcagtac aatgagaccg gcttctaatc aagtcccacg tgtcatgtca
     1501 gctcaacgtg ttgccaatac atcaacccag actatgggtc ctcgtccaac gactgctgct
     1561 gcagctgcag catctgctgt cagggctgtt cctcagtata aatatgcggc gggtgtacgt
     1621 aatcagcagc atcttaatac acagccccaa gtggctatgc agcagcctgc tgtccatgta
     1681 caaggccagg aacctctgac ggcttccatg ctggcagctg ctccacctca agagcagaaa
     1741 caaatgctag gtgagcggct cttcccactt atccaagcaa tgcacccaac tctggctggc
     1801 aaaattactg gaatgcttct ggagattgac aactcggagc ttctccatat gcttgaatca
     1861 cccgagtcac tgcgcttaaa ggttgatgaa gctgttgctg tgcttcaagc ccatcaggct
     1921 aaggaagctg ctcagaaggt tgttaatgca actggagtgc caactgctga ttacaaggat
     1981 gacgacgata agtaaaccag cctcaagaac acccgaatgg agtctctaag ctacataata
     2041 ccaacttaca ctttacaaaa tgttgtcccc caaaatgtag ccattcgtat ctgctcctaa
     2101 taaaaagaaa gtttcttcac attctggcgt agatccagac atgaaaaaaa aaaaaaaaaa
     2161 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2221 aaaaaaaaaa aaaaaaaaaa aaacagtagg gttgtaaagg tttttctttt cctgagaaaa
     2281 caaccttttg ttttctcagg ttttgctttt tggcctttcc ctagctttaa aaaaaaaaaa
     2341 agcaaaagct agccatggtc ccagcctcct cgctggcggc tagtgggcaa catgcttcgg
     2401 catggcgaat gggacgcggc cgcatgcata agcttgagta ttctatagtg tcacctaaat
     2461 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2521 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2581 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2641 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     2701 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2761 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2821 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2881 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2941 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3001 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3061 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3121 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3181 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3241 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3301 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3361 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3421 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3481 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3541 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3601 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3661 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3721 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3781 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3841 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3901 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3961 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4021 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4081 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4141 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4201 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4261 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4321 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4381 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4441 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4501 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4561 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4621 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     4681 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     4741 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     4801 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     4861 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     4921 aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     4981 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     5041 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     5101 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     5161 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     5221 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     5281 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     5341 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc
     5401 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     5461 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     5521 gtcacgacgt tgtaaaacga cggccagtga attgtaatac gactcactat a
//
