LOCUS       sequence-343597-        5610 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Template for making mRNA for overexpressing frog PABPC1 M161A mutant
            (C-FLAG) in frog oocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5610)
  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 5610)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: C233
FEATURES             Location/Qualifiers
     source          1..5610
                     /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"
     5'UTR           33..75
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             1986..2009
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     3'UTR           2012..2137
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(2498..2516)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2499..2516)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2534..2550)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2534..2550)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2547..2569)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2558..2574
                     /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(2582..2588,2589..2606,2607..2612))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2627..2648
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2765..2782)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2936..3524)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3016..3035)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3695..4486,4487..4555))
                     /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     4318..4337
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4556..4660)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4728..4746
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4784..4806)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4906..4925
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4992..5447)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5129..5150)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5341..5360
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5573..5595
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5587..5604
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5588..5604
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg gaatacaagc ttcttgttct ttttgcagaa gctcagaata
       61 aacgctcaac tttggcagat ctgaacatga atcccagtgc tcccagctac ccaatggctt
      121 ccctgtacgt tggagacctt catcaagatg taacggaggc catgctttat gagaagttta
      181 gcccagctgg tcccattttg tcaatccgag tttgcagaga tatgattaca cgacgctcac
      241 ttggatatgc atatgtcaac ttccagcaac cagctgatgc tgaacgtgct ttggatacaa
      301 tgaactttga tgtcataaag ggcaggcccg tgcggattat gtggtctcag cgtgaccctt
      361 ctctgcgtaa gagtggagtt ggtaacattt tcatcaaaaa cctggacaag tctattgata
      421 acaaagcctt gtacgataca ttttctgcat ttggaaacat tctttcctgt aaggtggttt
      481 gtgatgaaaa tggatccaag ggctatggtt ttgttcactt tgagacacaa gaggctgctg
      541 agagggctat tgataaaatg aatggcgcgc ttctcaatga ccgcaaagta tttgttgggc
      601 gctttaagtc ccgcaaagag cgtgaagctg agcttggtgc cagagctaag gaatttacaa
      661 atgtttacat caaaaatttt ggagacgaca tgaatgacga gaggctcaaa gaaatgtttg
      721 gcaaatatgg gccagctctt agtgttaaag ttatgactga tgacaatgga aagtcaaaag
      781 gctttggctt tgtcagcttt gaaagacatg aagatgcaca aaaggctgtt gatgaaatgt
      841 atggcaagga tatgaatggg aagtccatgt ttgtgggccg tgcacagaaa aaggtggaaa
      901 ggcaaactga gcttaagcgc aagtttgaac aaatgaatca ggaccgaatc accagatacc
      961 agggtgttaa cctctatgtt aaaaaccttg atgatggtat tgatgatgaa cgactgcgga
     1021 aagaattctt accttttggt acaatcacca gtgctaaggt aatgatggaa ggtggtcgca
     1081 gtaaaggctt tggttttgta tgcttttctt cacctgaaga ggccactaaa gcagtcacag
     1141 aaatgaatgg tagaattgtt gccacaaaac ccctgtatgt tgcattggcc caaagaaaag
     1201 aagagcgtca ggctcacttg accaaccagt acatgcagag gatggcaagt gttcgtgtac
     1261 caaatcctgt gatcaaccca taccagccac caccatccag ctatttcatg gcagctatcc
     1321 caccggctca aaaccgtgct gcgtactacc cacctgggca gattgcacag ctcaggccca
     1381 gcccccgttg gactgcacag ggtgccagac ctcatccatt ccagaacatg cccggagcaa
     1441 tccgccctac tgctccaaga ccaccaacat tcagtacaat gagaccggct tctaatcaag
     1501 tcccacgtgt catgtcagct caacgtgttg ccaatacatc aacccagact atgggtcctc
     1561 gtccaacgac tgctgctgca gctgcagcat ctgctgtcag ggctgttcct cagtataaat
     1621 atgcggcggg tgtacgtaat cagcagcatc ttaatacaca gccccaagtg gctatgcagc
     1681 agcctgctgt ccatgtacaa ggccaggaac ctctgacggc ttccatgctg gcagctgctc
     1741 cacctcaaga gcagaaacaa atgctaggtg agcggctctt cccacttatc caagcaatgc
     1801 acccaactct ggctggcaaa attactggaa tgcttctgga gattgacaac tcggagcttc
     1861 tccatatgct tgaatcaccc gagtcactgc gcttaaaggt tgatgaagct gttgctgtgc
     1921 ttcaagccca tcaggctaag gaagctgctc agaaggttgt taatgcaact ggagtgccaa
     1981 ctgctgatta caaggatgac gacgataagt aaaccagcct caagaacacc cgaatggagt
     2041 ctctaagcta cataatacca acttacactt tacaaaatgt tgtcccccaa aatgtagcca
     2101 ttcgtatctg ctcctaataa aaagaaagtt tcttcacatt ctggcgtaga tccagacatg
     2161 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2221 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2281 aaaaaaaaaa aaaaaaaaac agtagggttg taaaggtttt tcttttcctg agaaaacaac
     2341 cttttgtttt ctcaggtttt gctttttggc ctttccctag ctttaaaaaa aaaaaaagca
     2401 aaagctagcc atggtcccag cctcctcgct ggcggctagt gggcaacatg cttcggcatg
     2461 gcgaatggga cgcggccgca tgcataagct tgagtattct atagtgtcac ctaaatagct
     2521 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2581 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2641 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2701 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2761 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2821 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2881 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2941 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3001 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3061 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3121 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3181 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3241 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3301 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3361 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3421 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3481 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3541 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3601 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3661 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3721 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3781 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3841 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3901 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3961 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4021 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4081 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4141 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4201 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4261 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4321 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4381 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4441 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4501 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4561 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4621 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4681 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     4741 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     4801 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     4861 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     4921 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     4981 accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     5041 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     5101 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     5161 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     5221 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     5281 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     5341 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     5401 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc
     5461 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     5521 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     5581 cgacgttgta aaacgacggc cagtgaattg
//
