LOCUS       sequence-343308-        5559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for making mRNA for overexpressing frog ePAB (C-FLAG) in 
            frog oocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5559)
  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 5559)
  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: C078
FEATURES             Location/Qualifiers
     source          1..5559
                     /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             1957..1980
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     3'UTR           1983..2108
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(2430..2448)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2431..2448)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2466..2482)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2466..2482)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2479..2501)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2490..2506
                     /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(2514..2520,2521..2538,2539..2544))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2559..2580
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2697..2714)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2868..3456)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2948..2967)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3627..4418,4419..4487))
                     /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     4250..4269
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4488..4592)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4660..4678
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4716..4738)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4838..4857
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4924..5379)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5061..5082)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5273..5292
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5505..5527
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5519..5536
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5520..5536
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5543..5559,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(5543..5559,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaataca agcttcttgt tctttttgca gaagctcaga ataaacgctc aactttggca
       61 gatctgaaca tgaacgcaac cagagccgaa tacccgctgg catctcttta tataggggac
      121 ctccaccccg acgtgacgga agcgatgctg tatgagaagt tttctccagc ggggcccatc
      181 atgtctataa gagtgtgtcg agatattgct actcgccggt cccttggcta tgcctacatt
      241 aatttccagc agccggctga tgctgagcgg gccttggata caatgaattt tgaggttatt
      301 aaaggcaggc caatccgaat catgtggtcc cagagagatc cgggccttcg aaaatcgggt
      361 gtagggaatg tatttattaa aaatttagac gactccatcg acaacaaagc cctgtatgac
      421 actttctcag cgtttggaga tattttatct tgtaaggtgg tttgcgatga atacggatct
      481 cgaggttatg gtttcgtaca ttttgagacc caagaagctg ccaatcgagc gattcagact
      541 atgaatggga tgttgctcaa cgaccgcaaa gtgtttgtgg gccattttaa atcacggagg
      601 gaaagagaat tagaatatgg ggcaaaagtc atggaattta ccaacgtgta catcaagaac
      661 ttcggggagg atatggacga taaaagactt aaagaaatat tttctgcctt tggaaatacg
      721 ttaagcgtta aagtaatgat ggataatagt ggtcgatccc ggggttttgg ttttgttaat
      781 tatggaaatc acgaagaagc acaaaaagca gttactgaga tgaacggtaa agaagtaaac
      841 ggtcgaatgg tttacgtagg cagagctcag aagaggatag aacggcaggg cgagttaaaa
      901 cgcaagttcg aacaaatcaa gcaagaacga atcaaccgct atcagggtgt taatttgtat
      961 gttaaaaacc tggacgatgg tatcgatgat gaccggctca gaaaggagtt ctcgccgtat
     1021 ggtaccatta ccagtacaaa ggtaatgact gaaggaggcc atagtaaagg ctttggtttt
     1081 gtttgcttct cctccccaga agaagccact aaagcagtca cagaaatgaa cgggaggata
     1141 gtcagtacga agcctctata tgttgcactt gcacaaagga aggaagaacg gaaggcaatt
     1201 cttaccaatc agtacatgca gagactagcc actatgaggg cgatgcctgg tcctctcctt
     1261 gggtccttcc agcaacctgc taattatttc ttgcctacaa tgccacagcc ttcaaacaga
     1321 gcattttata gtccaaatcc tgttgctcct gttcgtccag ctcctcagtg ggcttcccac
     1381 cagtccaggc ctccacagta ccaaccacca gccccattaa tgagggcggt gccaccaaga
     1441 aggatgcatt caaatatcag tacaatgaag caggcatcaa cacaagtacc cagggtgcct
     1501 ctgcagagtc aaagagtagc aaacattggc actcaaacag ctggagctcg ggcccaagta
     1561 aacgcgtcca taatgcgcgc aatgccacat tacaagtatt cctgtggcgt gaggaacgtg
     1621 cagcctatag gaagctctgc tcatctgcaa caggtactgg aacctgcagt tctgatgcaa
     1681 ggacaagaac cgctaactgc atctctgctt gcagctgccc cactgcagga acagaaacaa
     1741 atactcgggg aacgtattta cccgttaata catgaaatgc atccgacgtt agccggaaag
     1801 ataacgggaa tgctgctgga aattgataac tctgagctac tgcatatgct ggagtcacca
     1861 gagtctctgc attcaaaggt ggaagaggca gtggcggtcc tgcaagccca ccaagccaag
     1921 gaaagtgctc cgaaaagtgc tccgcagtct ctgatcgatt acaaggatga cgacgataag
     1981 taaaccagcc tcaagaacac ccgaatggag tctctaagct acataatacc aacttacact
     2041 ttacaaaatg ttgtccccca aaatgtagcc attcgtatct gctcctaata aaaagaaagt
     2101 ttcttcacat tctggcgtag atccagacat gaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2161 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2221 aaaaaaaaaa acagtagggt tgtaaaggtt tttcttttcc tgagaaaaca accttttgtt
     2281 ttctcaggtt ttgctttttg gcctttccct agctttaaaa aaaaaaaaag caaaagctag
     2341 ccatggtccc agcctcctcg ctggcggcta gtgggcaaca tgcttcggca tggcgaatgg
     2401 gacgcggccg catgcataag cttgagtatt ctatagtgtc acctaaatag cttggcgtaa
     2461 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     2521 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     2581 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2641 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2701 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2761 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2821 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2881 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2941 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3001 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3061 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3121 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3181 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3241 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3301 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3361 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3421 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3481 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3541 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3601 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3661 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3721 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3781 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3841 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3901 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3961 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4021 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4081 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4141 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4201 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4261 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4321 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4381 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4441 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4501 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4561 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     4621 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     4681 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     4741 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     4801 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     4861 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     4921 aggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     4981 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     5041 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     5101 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     5161 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     5221 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     5281 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     5341 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg
     5401 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     5461 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     5521 taaaacgacg gccagtgaat tgtaatacga ctcactata
//
