LOCUS       sequence-257448-        4318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid of Anbu from Y. bercovieri (inactive mutant; 
            (UNIPROT A0A0T9RXH3 variant with T1A, V14A and G107H mutations).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4318)
  AUTHORS   Piasecka A, Czapinska H, Vielberg MT, Szczepanowski RH, Kiefersauer 
            R, Reed S, Groll M, Bochtler M
  TITLE     The Y. bercovieri Anbu crystal structure sheds light on the 
            evolution of highly (pseudo)symmetric multimers.
  JOURNAL   J Mol Biol. 2018 Mar 2;430(5):611-627. doi: 
            10.1016/j.jmb.2017.11.016. Epub 2017 Dec 16.
  PUBMED    29258816
REFERENCE   2  (bases 1 to 4318)
  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"; doi: 
            "10.1016/j.jmb.2017.11.016"; journalName: "J Mol Biol"; date: 
            "2018-03-2- 2"; volume: "430"; issue: "5"; pages: "611-627"
COMMENT     Sequence Label: pET20b-YbAnbuT1A
FEATURES             Location/Qualifiers
     source          1..4318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             819..836
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(889..907)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      903..950
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      987..1442
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1074..1093)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1284..1305
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1468..1572
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1573..1641,1642..2433)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1791..1810)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2604..3192
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3093..3112
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3346..3363
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3378..3517
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3443..3462)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3603..3625
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3619..3810)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atggcttact gtgtggccat gcggctgtcg tccgggttag
      121 cgtttgcttc cgactcccga accaacgccg gagtggatca tatttccact tttcgtaaat
      181 tgcatctgtt tcagcaaccg ggcgaacgaa cactggtggt gcaaagcgcg ggcaatttgg
      241 cgaccacaca gagtattgtc agcttgctgc aacgccgctg tctcgatcct gagcaaacca
      301 acttgatgaa tgtcgcctca atgtacgaag ccgccaccct gttgggtgaa acggtgcgtg
      361 aggtgattaa ccgggacagc ggcgcacagc aaaatagcca cggcaccgat tttaactgta
      421 acctgttgtt aggtggtcag attaagggtg aagggctacg gctgttccac atctatccgc
      481 agggtaattt tattgaagcc actcaggaca ccccctattt tcagattggc gagagtaaat
      541 atgggaagcc cattattgat cgcgtgttga gctacgacac cccactggat caggcgatgc
      601 aatgtgcact gatttcgatg gactcgactt tacgcagtaa tttatccgtg ggcttgccgc
      661 tggatgtgat gatctacccc ctcgacagtt tcagcaccga acagcagtat agaatcaccg
      721 aggatcaccc ctattttatg atgatccgta agggctgggg cgagggtttg gtgagtattt
      781 ttgcgcagtt accggggttg aaattgggga ggctcgagca ccaccaccac caccactgag
      841 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
      901 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
      961 gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     1021 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     1081 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     1141 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     1201 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     1261 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     1321 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     1381 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     1441 ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     1501 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     1561 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     1621 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     1681 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     1741 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     1801 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     1861 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     1921 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     1981 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     2041 gtaactcgcc ttgatcgctg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     2101 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     2161 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     2221 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     2281 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     2341 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     2401 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     2461 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     2521 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     2581 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2641 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2701 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     2761 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2821 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2881 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg tccgtgcaca
     2941 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3001 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3061 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3121 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3181 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3241 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3301 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3361 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3421 caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     3481 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     3541 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3601 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     3661 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     3721 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     3781 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     3841 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     3901 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     3961 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
     4021 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
     4081 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
     4141 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
     4201 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
     4261 gatcatgcgc acccgtggcc aggacccaac gctgcccgag atctcgatcc cgcgaaat
//
