LOCUS       sequence-423116-        5768 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid encodes a protein (Non tagged Avidin) that is used for 
            generating functionalized magnetic beads for MagIC-cryo-EM, a method
            for single-particle cryo-EM analysis on magnetic beads..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5768)
  AUTHORS   Arimura Y, Konishi HA, Funabiki H
  TITLE     MagIC-Cryo-EM, structural determination on magnetic beads for scarce
            macromolecules in heterogeneous samples.
  JOURNAL   eLife. 2025 May 20;13:RP103486. doi: 10.7554/eLife.103486.
  PUBMED    40390365
REFERENCE   2  (bases 1 to 5768)
  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: "2025-05-20"; volume: "13:RP103486. doi"; pages: " 
            10.7554/eLife.103486"
COMMENT     Sequence Label: Non tagged Avidin
FEATURES             Location/Qualifiers
     source          1..5768
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     613..632
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(652..729)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     935..954
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1038..1057
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1038..1056
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1057..1081
                     /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             1096..1118
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1537..1554
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1607..1625)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1621..1668
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1705..2160
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1792..1811)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2002..2023
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2186..2290
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2291..2359,2360..3151)
                     /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(2509..2528)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3322..3910
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3811..3830
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4064..4081
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4096..4235
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4161..4180)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4321..4343
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4337..4528)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5337..5768,1..651))
                     /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 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
       61 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
      121 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
      181 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
      241 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
      301 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
      361 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
      421 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
      481 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
      541 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
      601 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
      661 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
      721 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
      781 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
      841 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
      901 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
      961 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     1021 tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc
     1081 ccctctagaa ataattttgt ttaactttaa gaaggagata tacatatggc tgaagctggt
     1141 atcaccggca cctggtacaa ccagctggga tccaccttca tcgttaccgc tggtgctgac
     1201 ggtgctctga ccggtaccta cgaatccgct gttggtaacg ctgaatctag atacgttctg
     1261 accggtcgtt acgactccgc tccggctacc gacggttccg gaaccgctct gggttggacc
     1321 gttgcttgga aaaacaacta ccgtaacgct cactccgcta ccacctggtc tggccagtac
     1381 gttggtggtg ctgaagctcg tatcaacacc cagtggttgt tgacctccgg caccaccgaa
     1441 gccaacgcgt ggaaatccac cctggttggt cacgacacct tcaccaaagt taaaccgtcc
     1501 gctgcttcct aataaaagct tgcggccgca ctcgagcacc accaccacca ccactgagat
     1561 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     1621 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     1681 actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     1741 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     1801 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     1861 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     1921 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     1981 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2041 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2101 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     2161 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2221 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2281 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2341 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2401 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     2461 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     2521 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2581 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2641 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     2701 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2761 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2821 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2881 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2941 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3001 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3061 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3121 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3181 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3241 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3301 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3361 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3421 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     3481 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3541 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3601 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3661 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3721 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3781 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3841 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3901 cctatggaaa aacgccagca acgcggcctt ttcacggttc ctggcctttt gctggccttt
     3961 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4021 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4081 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4141 ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     4201 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     4261 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4321 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     4381 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     4441 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     4501 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     4561 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
     4621 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg
     4681 accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc
     4741 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg
     4801 acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc
     4861 aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat
     4921 tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga
     4981 tcatgcgcac ccgtggggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt
     5041 tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa
     5101 gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga
     5161 gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga
     5221 cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     5281 tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac
     5341 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5401 cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg
     5461 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
     5521 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
     5581 gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg
     5641 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
     5701 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
     5761 cagtcgcc
//
