LOCUS       sequence-343588-        6321 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Repair plasmid for introducing AID, T2A and mCherry at the C-teminus
            of human PABPC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6321)
  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 6321)
  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: C152
FEATURES             Location/Qualifiers
     source          1..6321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(16..444)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(126..147)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     338..357
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        471..575
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(576..644,645..1436)
                     /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     complement(794..813)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1607..2195
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2096..2115
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2349..2366
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2483..2504
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2519..2524,2525..2542,2543..2549)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2557..2573
                     /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)."
     primer_bind     2562..2584
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2581..2597
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2581..2597
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2618..2636
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             3696..4376
                     /codon_start=1
                     /gene="Arabidopsis thaliana AXR3"
                     /product="auxin-responsive IAA17 protein"
                     /label=auxin-inducible degron
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="GSVELNLRETELCLGLPGGDTVAPVTGNKRGFSETVDLKLNLNNE
                     PANKEGSTTHDVVTFDSKEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGG
                     PEAAAFVKVSMDGAPYLRKIDLRMYKSYDELSNALSNMFSSFTMGKHGGEEGMIDFMNE
                     RKLMDLVNSWDYVPSYEDKDGDWMLVGDVPWPMFVDTCKRLRLMKGSDAIGLAPRAMEK
                     CKSRA"
     CDS             4377..4430
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4443..5153
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4588..4606)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4856..4875
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(6127..6146)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6128..6146)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6153..6170)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6153..6169)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6162..6184)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
       61 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
      121 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
      181 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
      241 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
      301 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
      361 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
      421 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
      481 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
      541 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
      601 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
      661 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
      721 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
      781 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
      841 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
      901 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
      961 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     1021 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     1081 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     1141 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     1201 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     1261 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     1321 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     1381 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     1441 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1501 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1561 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1621 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1681 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1741 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1801 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1861 agtcgtgtct taccgggttg gacccaagac gatagttacc ggataaggcg cagcggtcgg
     1921 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1981 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2041 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2101 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2161 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2221 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2281 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     2341 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     2401 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     2461 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     2521 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     2581 caggaaacag ctatgaccat gattacgcca agctcgaaat taaccctcac taaagggaac
     2641 aaaagctggt accgggcccc ccgaacaccg atttaacaac ctaatggtaa cattgcttaa
     2701 ttggcaagct gataatatca agagttaatt gccttgtggc agattttatt ctgtcttccc
     2761 atgaagacat aatgaaattg tggattatgg gttattatat acgtgagaca gaagtatggt
     2821 tcttaaaccc cagtttaatg tggtaattca atgcagttaa aaactttttt ttattgaagg
     2881 gagtttttga atgtttctgt tcctaggtga acggctgttt cctcttattc aagccatgca
     2941 ccctactctt gctggtaaaa tcactggcat gttgttggag attgataatt cagaacttct
     3001 tcatatgctc gagtctccag agtcactccg ttctaaggtg atttaaattt atttaccaag
     3061 ttgcctctta cacagttatt acgtggcttt ttatggggac ttaattgtac agaacactta
     3121 catcgtgtta tatataagcc atgtacagtt gtaaagtctg cataataacc tgtggcgcta
     3181 tttatcccca ttttatagat aggtaaactg aaacaggctg aataatttgc caaatgctca
     3241 cacagctctt aagtggcaga gcctggcttc aaatcccatc agctccaaca gccatactct
     3301 caactgctgc tgctttctct agtggagaaa gagaaatatt tttcgagtgt ggggaggaca
     3361 aggaaagagt agtgcagggg agcgattcat aattctagaa tagtggtgta atagaactca
     3421 aaattgaatg gatatagttg tttagtgtaa caatttatac tttcatttga gggaagaaac
     3481 ttatctttgt tgggaattgg tatgttacca ttatgaaact ctttatttcc attcctttcc
     3541 tgaaccagtt tatctgtttt cccctttttt aggttgatga agctgtagct gtactacaag
     3601 cccaccaagc taaagaggct gcccagaaag cagttaacag tgccaccggt gttccaactg
     3661 ttgtcgactc cggagccgcc gctgctggcg gatctggcag tgtcgagctg aatctgaggg
     3721 agactgagct gtgtcttggt cttcccggtg gagatacagt ggctccggta accggaaaca
     3781 agagagggtt ctcagagacg gttgatctga agctaaatct gaataatgag cctgcaaaca
     3841 aggaaggatc tacgactcat gacgtcgtga cttttgattc caaggagaag agtgcttgtc
     3901 ctaaagatcc agccaaacct ccggccaagg cacaagttgt gggatggcca ccggtgagat
     3961 cataccggaa gaacgtgatg gtttcctgcc aaaaatcaag cggtggcccg gaggcggcgg
     4021 cgttcgtgaa ggtatcaatg gacggagcac cgtacttgag gaaaatcgat ttgaggatgt
     4081 ataaaagcta cgatgagctt tctaatgctt tgtccaacat gttcagctct tttaccatgg
     4141 gcaaacatgg aggagaagaa ggaatgatag acttcatgaa tgagaggaaa ttgatggatt
     4201 tggtgaatag ctgggactat gttccctctt atgaagacaa agacggtgat tggatgctcg
     4261 tcggcgacgt tccttggcca atgttcgtcg atacatgcaa gcgtttacgt ctcatgaaag
     4321 gatcggatgc cattggtctc gctccgaggg cgatggagaa gtgcaagagc agagctgagg
     4381 gcagaggaag tctgctaaca tgcggtgacg tcgaggagaa tcctggccca gcggccgcaa
     4441 ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     4501 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     4561 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     4621 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     4681 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     4741 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tctctccagg
     4801 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     4861 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     4921 gcgctctgaa aggagagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     4981 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     5041 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     5101 aacgcgccga gggacggcac tccaccggag gaatggacga gctgtacaaa taaagtgagc
     5161 ttttccctct ggtttgttgt tttaagcttg ttttcttcat cttttataaa gtttgttgag
     5221 catattttga aatgtacata gcaaaagatc attttacagt tcatttaaat gtattgctaa
     5281 tttttcaagg aaaagtggaa ctaactatag aggaattaaa ggaattttac ccgttaactg
     5341 ttttattttt aaaatcccag gactttcagg tggtgttttt tttttttttt tgagatggag
     5401 tttcgttctg tcgcccaacc tggagtgcat tgacgcgatc tcggctcact gcaacctcca
     5461 cctcccaggt tcaagcaatt ctcctgcctc ggcctcccga gcagctggga ttaacaggcc
     5521 catgcctgcc accgtgccca gctaattttt gtatttttac tagacttggg gttttgccat
     5581 gttggccagg ctggtctgga acttctgacc tcaagtgatc cgcccacctc ggcctcccaa
     5641 agtcttgaaa ttacaggcgt gagccactgc gccctgccag attttgtata gtaatatata
     5701 tataaaatga cctgtcatag gttggctttc atagttttac ctttcctttt tgggtggtgc
     5761 ccttgggtct ctactactgt ctttcatcac tcccaagaaa ttgagtagtg cagccaggat
     5821 caggaccagg gattggcaaa tgttttttgt aaagggccag ataggaaata ttttgggctt
     5881 tgaggactgt gttatctctg aaatgatctt gttccagtgt taccatgtca agacaaccat
     5941 agatagtatg tatacaaatg aacaggtatg gctgtgttcc agtaaaacta tggataaaaa
     6001 caaccacatt gcatttggct aagggcttta atttgctact gacaagtgat agtcctggtc
     6061 ttaatttgtt tttttaacat ttacagattg atcagggacc atgcgccgcg gtggagctcc
     6121 aattcgccct atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac
     6181 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     6241 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     6301 ggcgaatgga aattgtaagc g
//
