LOCUS       sequence-343508-        7134 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for expressing frog PABPC1 (C-His) in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7134)
  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 7134)
  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: C064
FEATURES             Location/Qualifiers
     source          1..7134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     965..984
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1004..1081)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1287..1306
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1390..1409
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1390..1408
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1409..1433
                     /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)."
     RBS             1448..1470
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3388..3405
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3458..3476)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3472..3519
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3556..4011
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3643..3662)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3853..3874
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4103..4918)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4103..4918)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4826..4845
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4910..4929
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5040..5628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5529..5548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5782..5799
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5814..5953
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5879..5898)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6039..6061
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6055..6246)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7055..7134,1..1003))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
       61 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
      121 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
      181 cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc
      241 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
      301 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
      361 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
      421 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
      481 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
      541 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
      601 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
      661 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
      721 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
      781 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
      841 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
      901 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
      961 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     1021 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt
     1081 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     1141 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     1201 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     1261 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     1321 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
     1381 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
     1441 aaataatttt gtttaacttt aagaaggaga tataccatga atcccagtgc tcccagctac
     1501 ccaatggctt ccctgtacgt tggagacctt catcaagatg taacggaggc catgctttat
     1561 gagaagttta gcccagctgg tcccattttg tcaatccgag tttgcagaga tatgattaca
     1621 cgacgctcac ttggatatgc atatgtcaac ttccagcaac cagctgatgc tgaacgtgct
     1681 ttggatacaa tgaactttga tgtcataaag ggcaggcccg tgcggattat gtggtctcag
     1741 cgtgaccctt ctctgcgtaa gagtggagtt ggtaacattt tcatcaaaaa cctggacaag
     1801 tctattgata acaaagcctt gtacgataca ttttctgcat ttggaaacat tctttcctgt
     1861 aaggtggttt gtgatgaaaa tggatccaag ggctatggtt ttgttcactt tgagacacaa
     1921 gaggctgctg agagggctat tgataaaatg aatggcatgc ttctcaatga ccgcaaagta
     1981 tttgttgggc gctttaagtc ccgcaaagag cgtgaagctg agcttggtgc cagagctaag
     2041 gaatttacaa atgtttacat caaaaatttt ggagacgaca tgaatgacga gaggctcaaa
     2101 gaaatgtttg gcaaatatgg gccagctctt agtgttaaag ttatgactga tgacaatgga
     2161 aagtcaaaag gctttggctt tgtcagcttt gaaagacatg aagatgcaca aaaggctgtt
     2221 gatgaaatgt atggcaagga tatgaatggg aagtccatgt ttgtgggccg tgcacagaaa
     2281 aaggtggaaa ggcaaactga gcttaagcgc aagtttgaac aaatgaatca ggaccgaatc
     2341 accagatacc agggtgttaa cctctatgtt aaaaaccttg atgatggtat tgatgatgaa
     2401 cgactgcgga aagaattctt accttttggt acaatcacca gtgctaaggt aatgatggaa
     2461 ggtggtcgca gtaaaggctt tggttttgta tgcttttctt cacctgaaga ggccactaaa
     2521 gcagtcacag aaatgaatgg tagaattgtt gccacaaaac ccctgtatgt tgcattggcc
     2581 caaagaaaag aagagcgtca ggctcacttg accaaccagt acatgcagag gatggcaagt
     2641 gttcgtgtac caaatcctgt gatcaaccca taccagccac caccatccag ctatttcatg
     2701 gcagctatcc caccggctca aaaccgtgct gcgtactacc cacctgggca gattgcacag
     2761 ctcaggccca gcccccgttg gactgcacag ggtgccagac ctcatccatt ccagaacatg
     2821 cccggagcaa tccgccctac tgctccaaga ccaccaacat tcagtacaat gagaccggct
     2881 tctaatcaag tcccacgtgt catgtcagct caacgtgttg ccaatacatc aacccagact
     2941 atgggtcctc gtccaacgac tgctgctgca gctgcagcat ctgctgtcag ggctgttcct
     3001 cagtataaat atgcggcggg tgtacgtaat cagcagcatc ttaatacaca gccccaagtg
     3061 gctatgcagc agcctgctgt ccatgtacaa ggccaggaac ctctgacggc ttccatgctg
     3121 gcagctgctc cacctcaaga gcagaaacaa atgctaggtg agcggctctt cccacttatc
     3181 caagcaatgc acccaactct ggctggcaaa attactggaa tgcttctgga gattgacaac
     3241 tcggagcttc tccatatgct tgaatcaccc gagtcactgc gcttaaaggt tgatgaagct
     3301 gttgctgtgc ttcaagccca tcaggctaag gaagctgctc agaaggttgt taatgcaact
     3361 ggagtgccaa ctgctgccgc actcgagcac caccaccacc accactgaga tccggctgct
     3421 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     3481 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     3541 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3601 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3661 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3721 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3781 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3841 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3901 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3961 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     4021 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4081 gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt
     4141 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     4201 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     4261 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     4321 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     4381 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     4441 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     4501 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     4561 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     4621 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     4681 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     4741 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     4801 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     4861 ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa
     4921 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgac caaaatccct
     4981 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5041 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5101 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5161 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     5221 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5281 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5341 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5401 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5461 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5521 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5581 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5641 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5701 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     5761 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     5821 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     5881 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     5941 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     6001 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     6061 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     6121 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     6181 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     6241 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     6301 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     6361 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     6421 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     6481 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     6541 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     6601 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     6661 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     6721 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     6781 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     6841 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     6901 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     6961 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     7021 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7081 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gttt
//
