LOCUS       sequence-343603-        8490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for making mRNA for overexpressing human eIF4G (C-FLAG) in 
            frog oocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8490)
  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 8490)
  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: C150
FEATURES             Location/Qualifiers
     source          1..8490
                     /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             4888..4911
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     3'UTR           4914..5039
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(5361..5379)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5362..5379)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5397..5413)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5397..5413)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5410..5432)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5421..5437
                     /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(5445..5451,5452..5469,5470..5475))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5490..5511
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5628..5645)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5799..6387)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5879..5898)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6558..7349,7350..7418))
                     /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     7181..7200
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7419..7523)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7591..7609
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7647..7669)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7769..7788
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7855..8310)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7992..8013)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8204..8223
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8436..8458
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8450..8467
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8451..8467
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(8474..8490,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(8474..8490,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaataca agcttcttgt tctttttgca gaagctcaga ataaacgctc aactttggca
       61 gatctgaaca tgaacaaagc tccacagtcc acaggccccc cacccgcccc atcccccgga
      121 ctcccacagc cagcgtttcc cccggggcag acagcgccgg tggtgttcag tacgccacaa
      181 gcgacacaaa tgaacacgcc ttctcagccc cgccagggag gattcaggtc tctgcagcac
      241 ttctacccta gccgggccca gcccccgagc agtgcagcct cccgagtgca gagtgcagcc
      301 cctgcccgcc ctggcccagc tgcccatgtc taccctgctg gatcccaagt aatgatgatc
      361 ccttcccaga tctcctaccc agcctcccag ggggcctact acatccctgg acaggggcgt
      421 tccacatacg ttgtcccgac acagcagtac cctgtgcagc caggagcccc aggcttctat
      481 ccaggtgcaa gccctacaga atttgggacc tacgctggcg cctactatcc agcccaaggg
      541 gtgcagcagt ttcccactgg cgtggccccc gccccagttt tgatgaacca gccaccccag
      601 attgctccca agagggagcg taagacgatc cgaattcgag atccaaacca aggaggaaag
      661 gatatcacag aggagatcat gtctggggcc cgcactgcct ccacacccac ccctccccag
      721 acgggaggcg gtctggagcc tcaagctaat ggggagacgc cccaggttgc tgtcattgtc
      781 cggccagatg accggtcaca gggagcaatc attgctgacc ggccagggct gcctggccca
      841 gagcatagcc cttcagaatc ccagccttcg tcgccttctc cgaccccatc accatcccca
      901 gtcttggaac cggggtctga gcctaatctc gcagtcctct ctattcctgg ggacactatg
      961 acaactatac aaatgtctgt agaagaatca acccccatct cccgtgaaac tggggagcca
     1021 tatcgcctct ctccagaacc cactcctctc gccgaaccca tactggaagt agaagtgaca
     1081 cttagcaaac cggttccaga atctgagttt tcttccagtc ctctccaggc tcccacccct
     1141 ttggcatctc acacagtgga aattcatgag cctaatggca tggtcccatc tgaagatctg
     1201 gaaccagagg tggagtcaag cccagagctt gctcctcccc cagcttgccc ctccgaatcc
     1261 cctgtgccca ttgctccaac tgcccaacct gaggaactgc tcaacggagc cccctcgcca
     1321 ccagctgtgg acttaagccc agtcagtgag ccagaggagc aggccaagga ggtgacagca
     1381 tcagtggcgc cccccaccat cccctctgct actccagcta cggctccttc agctacttcc
     1441 ccagctcagg aggaggaaat ggaagaagaa gaagaagagg aagaaggaga agcaggagaa
     1501 gcaggagaag ctgagagtga gaaaggagga gaggaactgc tccccccaga gagtacccct
     1561 attccagcca acttgtctca gaatttggag gcagcagcag ccactcaagt ggcagtatct
     1621 gtgccaaaga ggagacggaa aattaaggag ctaaataaga aggaggctgt tggagacctt
     1681 ctggatgcct tcaaggaggc gaacccggca gtaccagagg tggaaaatca gcctcctgca
     1741 ggcagcaatc caggcccaga gtctgagggc agtggtgtgc ccccacgtcc tgaggaagca
     1801 gatgagacct gggactcaaa ggaagacaaa attcacaatg ctgagaacat ccagcccggg
     1861 gaacagaagt atgaatataa gtcagatcag tggaagcctc taaacctaga ggagaaaaaa
     1921 cgttacgacc gtgagttcct gcttggtttt cagttcatct ttgccagtat gcagaagcca
     1981 gagggattgc cacatatcag tgacgtggtg ctggacaagg ccaataaaac accactgcgg
     2041 ccactggatc ccactagact acaaggcata aattgtggcc cagacttcac tccatccttt
     2101 gccaaccttg gccggacaac ccttagcacc cgtgggcccc caaggggtgg gccaggtggg
     2161 gagctgcccc gtgggccggc tggcctggga ccccggcgct ctcagcaggg accccgaaaa
     2221 gaaccacgca agatcattgc cacagtgtta atgaccgaag atataaaact gaacaaagca
     2281 gagaaagcct ggaaacccag cagcaagcgg acggcggctg ataaggatcg aggggaagaa
     2341 gatgctgatg gcagcaaaac ccaggaccta ttccgcaggg tgcgctccat cctgaataaa
     2401 ctgacacccc agatgttcca gcagctgatg aagcaagtga cgcagctggc catcgacacc
     2461 gaggaacgcc tcaaaggggt cattgacctc atttttgaga aggccatttc agagcccaac
     2521 ttctctgtgg cctatgccaa catgtgccgc tgcctcatgg cgctgaaagt gcccactacg
     2581 gaaaagccaa cagtgactgt gaacttccga aagctgttgt tgaatcgatg tcagaaggag
     2641 tttgagaaag acaaagatga tgatgaggtt tttgagaaga agcaaaaaga gatggatgaa
     2701 gctgctacgg cagaggaacg aggacgcctg aaggaagagc tggaagaggc tcgggacata
     2761 gcccggcggc gctctttagg gaatatcaag tttattggag agttgttcaa actgaagatg
     2821 ttaacagagg caataatgca tgactgtgtg gtcaaactgc ttaagaacca tgatgaagag
     2881 tcccttgagt gcctttgtcg tctgctcacc accattggca aagacctgga ctttgaaaaa
     2941 gccaagcccc gaatggatca gtatttcaac cagatggaaa aaatcattaa agaaaagaag
     3001 acgtcatccc gcatccgctt tatgctgcag gacgtgctgg atctgcgagg gagcaattgg
     3061 gtgccacgcc gaggggatca gggtcccaag accattgacc agatccataa ggaggctgag
     3121 atggaagaac atcgagagca catcaaagtg cagcagctca tggccaaggg cagtgacaag
     3181 cgtcggggcg gtcctccagg ccctcccatc agccgtggac ttccccttgt ggatgatggt
     3241 ggctggaaca cagttcccat cagcaaaggt agccgcccca ttgacacctc acgactcacc
     3301 aagatcacca agcctggctc catcgattct aacaaccagc tctttgcacc tggagggcga
     3361 ctgagctggg gcaagggcag cagcggaggc tcaggagcca agccctcaga cgcagcatca
     3421 gaagctgctc gcccagctac tagtactttg aatcgcttct cagcccttca acaagcggta
     3481 cccacagaaa gcacagataa tagacgtgtg gtgcagagga gtagcttgag ccgagaacga
     3541 ggcgagaaag ctggagaccg aggagaccgc ctagagcgga gtgaacgggg aggggaccgt
     3601 ggggaccggc ttgatcgtgc gcggacacct gctaccaagc ggagcttcag caaggaagtg
     3661 gaggagcgga gtagagaacg gccctcccag cctgaggggc tgcgcaaggc agctagcctc
     3721 acggaggatc gggaccgtgg gcgggatgcc gtgaagcgag aagctgccct acccccagtg
     3781 agccccctga aggcggctct ctctgaggag gagttagaga agaaatccaa ggctatcatt
     3841 gaggaatatc tccatctcaa tgacatgaaa gaggcagtcc agtgcgtgca ggagctggcc
     3901 tcaccctcct tgctcttcat ctttgtacgg catggtgtcg agtctacgct ggagcgcagt
     3961 gccattgctc gtgagcatat ggggcagctg ctgcaccagc tgctctgtgc tgggcatctg
     4021 tctactgctc agtactacca agggttgtat gaaatcttgg aattggctga ggacatggaa
     4081 attgacatcc cccatgtgtg gctctaccta gcggaactgg taacacccat tctgcaggaa
     4141 ggtggggtgc ccatggggga gctgttcagg gagattacaa agcctctgag accgttgggc
     4201 aaagctgctt ccctgttgct ggagatcctg ggcctcctgt gcaaaagcat gggtcctaaa
     4261 aaggtgggga cgctgtggcg agaagccggg cttagctgga aggaatttct acctgaaggc
     4321 caggacattg gtgcattcgt cgctgaacag aaggtggagt ataccctggg agaggagtcg
     4381 gaagcccctg gccagagggc actcccctcc gaggagctga acaggcagct ggagaagctg
     4441 ctgaaggagg gcagcagtaa ccagcgggtg ttcgactgga tagaggccaa cctgagtgag
     4501 cagcagatag tatccaacac gttagttcga gccctcatga cggctgtctg ctattctgca
     4561 attatttttg agactcccct ccgagtggac gttgcagtgc tgaaagcgcg agcgaagctg
     4621 ctgcagaaat acctgtgtga cgagcagaag gagctacagg cgctctacgc cctccaggcc
     4681 cttgtagtga ccttagaaca gcctcccaac ctgctgcgga tgttctttga cgcactgtat
     4741 gacgaggacg tggtgaagga ggatgccttc tacagttggg agagtagcaa ggaccccgct
     4801 gagcagcagg gcaagggtgt ggcccttaaa tctgtcacag ccttcttcaa gtggctccgt
     4861 gaagcagagg aggagtctga ccacaacgat tacaaggatg acgacgataa gtaaaccagc
     4921 ctcaagaaca cccgaatgga gtctctaagc tacataatac caacttacac tttacaaaat
     4981 gttgtccccc aaaatgtagc cattcgtatc tgctcctaat aaaaagaaag tttcttcaca
     5041 ttctggcgta gatccagaca tgaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     5101 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     5161 aacagtaggg ttgtaaaggt ttttcttttc ctgagaaaac aaccttttgt tttctcaggt
     5221 tttgcttttt ggcctttccc tagctttaaa aaaaaaaaaa gcaaaagcta gccatggtcc
     5281 cagcctcctc gctggcggct agtgggcaac atgcttcggc atggcgaatg ggacgcggcc
     5341 gcatgcataa gcttgagtat tctatagtgt cacctaaata gcttggcgta atcatggtca
     5401 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5461 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5521 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5581 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5641 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5701 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5761 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5821 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5881 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5941 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6001 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6061 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6121 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6181 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6241 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6301 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6361 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6421 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6481 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6541 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6601 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6661 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6721 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6781 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6841 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6901 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6961 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7021 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7081 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7141 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7201 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7261 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7321 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7381 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7441 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7501 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     7561 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     7621 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     7681 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     7741 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     7801 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg
     7861 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     7921 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     7981 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     8041 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     8101 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     8161 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     8221 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     8281 ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg
     8341 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     8401 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     8461 ggccagtgaa ttgtaatacg actcactata
//
