LOCUS       sequence-479510-        2965 bp    DNA     circular SYN 23-JUL-2025
DEFINITION  bacterial expression of human TTR mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2965)
  AUTHORS   Sun X, Ferguson JA, Yang K, Stanfield RL, Dyson HJ, Wright PE
  TITLE     Mispacking of the F87 sidechain drives aggregation-promoting 
            conformational fluctuations in the subunit interfaces of the 
            transthyretin tetramer.
  JOURNAL   Protein Sci. 2024 Sep;33(9):e5101. doi: 10.1002/pro.5101.
  PUBMED    39149996
REFERENCE   2  (bases 1 to 2965)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein 
            Sci."; date: "2024-09"; volume: "33(9):e5101. doi"; pages: " 
            10.1002/pro.5101"
COMMENT     Sequence Label: TTR A120S
FEATURES             Location/Qualifiers
     source          1..2965
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             530..552
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    567..591
                     /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     complement(591..610)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(592..610)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(711..728)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(882..1470)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(962..981)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1581..1600)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1592..2407
                     /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             1592..2407
                     /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     complement(1665..1684)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(2499..2954)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2636..2657)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2848..2867
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgtagcagcc ggatctcatt ccttgggatt ggtgacgaca gacgtggtgg aataggagta
      181 ggggctcagc agggcggcaa tggtgtagcg gcgggggccg gagtcgttgg ctgtgaatac
      241 cacctctgca tgctcatgga atggggagat gccaagtgcc ttccagtaag atttggtgtc
      301 tatttccact ttgtatatcc cttctacaaa ttcctcctca gttgtgagcc catgcagctc
      361 tccagactca ctggttttcc cagaggcaaa tggctcccag gtgtcatcag cagcctttct
      421 gaacacatgc acggccacat tgatggcagg actgcctcgg acagcatcta gaactttgac
      481 catcagagga cacttggatt caccggtgcc cgtaggaccc atatgtatat ctccttctta
      541 aagttaaaca aaattatttc tagaggggaa ttgttatccg ctcacaattc ccctatagtg
      601 agtcgtatta atttcgcggg atcgagatcg atctcgatcc tctacgccgg acgcatcgtg
      661 gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga ctcgctcact
      721 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
      781 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
      841 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      901 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      961 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1021 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1081 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1141 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1201 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1261 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1321 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1381 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1441 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1501 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgaacaa taaaactgtc
     1561 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg
     1621 ctctaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     1681 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
     1741 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     1801 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     1861 tcctgatgat gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt
     1921 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     1981 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     2041 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     2101 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     2161 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     2221 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     2281 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     2341 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     2401 tttctaagaa ttaattcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2461 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     2521 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     2581 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     2641 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     2701 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     2761 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     2821 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     2881 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     2941 gctacagggc gcgtcccatt cgcca
//
