LOCUS       sequence-400632-        6093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  His6 tagged Atg20 and untagged Atg24 from S. cerevisiae for 
            co-expression in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6093)
  AUTHORS   Reinhart EF, Katzenell S, Andhare D, Bauer KM, Ragusa MJ
  TITLE     A Comparative Analysis of the Membrane Binding and Remodeling 
            Properties of Two Related Sorting Nexin Complexes Involved in 
            Autophagy.
  JOURNAL   Biochemistry. 2022 Apr 14. doi: 10.1021/acs.biochem.2c00007.
  PUBMED    35421303
REFERENCE   2  (bases 1 to 6093)
  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: 
            "Biochemistry. 2022 Apr 14. doi: 10.1021/acs.biochem.2c00007."
COMMENT     Sequence Label: Atg20-24 pST39
FEATURES             Location/Qualifiers
     source          1..6093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    775..914
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(840..859)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1000..1022
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1016..1207)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1444..1463
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1444..1462
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1494..1516
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             2833..2855
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2875..2892
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2893..2913
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(4940..4958)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4954..5001
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        5068..5159
                     /gene="bla"
                     /label=AmpR promoter
     terminator      6024..6063
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
      841 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
      901 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
      961 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     1021 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     1081 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     1141 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     1201 tggtcactga tgcctccgtg taagggggat ttctgttcat ggggtaatga taccgatgaa
     1261 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     1321 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     1381 tcaatgccag cgcttcgtta atacagatgt aggtgttggc accatgcatc gatcccgcga
     1441 aattaatacg actcactata gggagaccac aacggtttcc ctctagaaat aattttgttt
     1501 aactttaaga aggagatata catatgacag acaaaggcaa gaacgattta acaagcaaag
     1561 ctaaggacaa agccagaggc aaccccgaga agccaccata ttggtttgaa atcattgttt
     1621 ctgatccaca aaaaagaact ggtgacccag gctcttcatc tggctacgtg agttatcaga
     1681 tatcaacaaa gactaacaac acgtcatttt atgataatcg tggcgatcca gaatctatta
     1741 tagttgtaca tagaagatac agtgacctat tgctcctgca tgatatttta ctcaacaggt
     1801 tccccacctg catcatcccg ccattacctg ataagaaagt ttttcaatac atagcaggcg
     1861 ataggtttag ccaaagattt acacaaaaga gatgtcatag tttgcaaaat ttcctaagaa
     1921 gggtctcttt gcatcccgat ctttctcaat caaaagtctt caaaactttt ttggtgagta
     1981 aagattggga atctcaccgc aaagtcctcc aggatagctt gcaaccaaat aaagatgaag
     2041 ttactgatgc ttttatgaat gccttcaaaa cagttcacaa gcaaaacgaa gaattcactg
     2101 agataaggga gaaaagtgat aaactagatc ggactgtgac aaagatagat aaactttttc
     2161 ataaggttgt gaaaaaaaat gattcgatgt ctgaagacta tacaaagttg ggcagtaatt
     2221 tacaggaatt acaagaattg gtcacaggcg aaaacgagga acttgctgcg aagttgaaga
     2281 tttttaatga aggtgtaaca cagctgtcat atggcttgca agatcttaca aagtatctgg
     2341 attacgaata tattgttgat ttgaaggatc ttgaacacta cattgatagt atgcgacagc
     2401 tgatcaaatt gaaagaccag aaacagatag actacgaaga actaagtgat tatttaacca
     2461 ggtcaatcaa agaaaaaaat aacttgattt ctggatacgg aggtagtaat ttctttgcca
     2521 ataaattgga agaactagcg ggaataaatc aggaggcttc acgtagagaa aaaatcaata
     2581 aactagaagg caaaataacg tcattaacag gtgaattaga aaacgctaaa aaggtggcgg
     2641 atggattcga acaggaatgc ttgaaggaaa tagaccattt cgaaagtgta aaaacggctg
     2701 aaatcaagaa atctctgggc tcactagcgg accatcatat cgaattctac gaaaggatac
     2761 ttgaagcatg ggaaaaagta gacgatagtc tataaaagct tcgccagggt tttcccagtc
     2821 gagctcaata attttgttta actttaagaa ggagatatac atatgggttc ttctcaccat
     2881 caccatcacc atgaaaacct gtacttccaa tccaatgcat cagacttaaa tgatgtccaa
     2941 gaaaatgcta aattgaattc ggaaactcgg aatacaggaa aggctgaacc acctcatggg
     3001 acaactgagt atgtggcaga ggcagaaatt tctaagaatg gagttggatc gccaaaaaag
     3061 agcccaaaaa agggtaaggt tggaaagggg gacaataaca aagtagaaac tgagttagta
     3121 cacacagccc tgttagaaaa agacaaccca ttcatggagg aggggccaac aggattcaca
     3181 aaatcagctt tattagaaat acccggtatg agaagccata acctaaagaa cccgaatgaa
     3241 gattacgagg atgactctga gggattattg cctttaaacc aagaatcgaa tgcagaaaca
     3301 tgtagaacaa gtcttagtgg aagtattaac tccatgaacg gagagacctc tgcgtcagaa
     3361 gaaccttctg taagtaatag aaagaagtcc gctaggatac acatacttga agcgaaaaga
     3421 gtttccgaag gtcaaggaag agcctacatc gcctatgtta ttcagtttga aaattctacc
     3481 gttcagagac gttacagcga tttcgagagt ttgagaagta tcttgataag attgtttcct
     3541 atgacgttga taccgccgat tccagaaaag cagtcaataa agaattatgg taagtcaata
     3601 actggttcga gttcaaagta tttgctgccc tcagaaggtt ctggttcagt agatctatca
     3661 ttgtctgtga tacatgcgtc ggtcaataat agtgatgaaa agttaatacg gcataggata
     3721 agaatgctca ctgagttctt gaacaaactt ttgacgaacg aagaaatcac aaaaacatct
     3781 ataattaccg attttttgga tcctaataac cataattggc atgaatttgt taacagttcg
     3841 tccacttttt catctttacc gaagagcatc ctccagtgta atccactaga tcccactaac
     3901 accactagaa ttcatgccat gttaccaata ccgggctcct cgtcacagtt attgttgaat
     3961 aaggagtcca acgacaaaaa aatggataag gaaagaagta agtcgtttac taatattgaa
     4021 caggattaca agcaatatga gaacttgtta gataacggga tttataagta taatagacga
     4081 accacgaaga cttatcacga tttgaaatcg gattacaatg aaatcggtga ggtatttgca
     4141 cagtttgccc atgaacaggc tcaagttggg gaattggccg aacagctttc ttacttaagc
     4201 aatgcctttt cggggtcctc gatttcttta gaaaaattag ttggaagact atactacaat
     4261 ataaatgagc ctttaaatga atctgttcat atggcaactt ctgcaagaga attgatcaaa
     4321 tacagaaaat tgaaatacct gcaaaatgaa atgattaaaa agtctctgaa ttccaaaaga
     4381 gcacagttgg agaagctgga agcgcaaaac aatgaatata aggatgttga caaaataatt
     4441 gacaacgaaa tgtctaaaag tcatactatt aatctggaac gccctaataa taatacaggt
     4501 agcggtggga aatcatatgg tggcaaacta ttcaatggat ttaacaaatt ggcatcaatg
     4561 gtaaaagatt ctgtgaagta tcaagaaaca gatccgcata ccgcaagtat caatttgaag
     4621 aaagaaattg aacaactgtc tgaatcattg gaagttactg agaatgattt ggaggtcatt
     4681 tcaaaagtca tcaaaaatga ccagctacct aaattttcta aagaaaggga agtagattta
     4741 tcagaaattt taaagcatta ttccagatat atgagaaatt atgcccgtca aaatttagaa
     4801 atttggaaag aagtcaaaag acatcaagat tttgcataag gtaccagcgg ataacaattt
     4861 cacatccgga ttcgcgaacg cgtctcgaga gatccggctg ctaacaaagc ccgaaaggaa
     4921 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     4981 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccgactagta gcccgcctaa
     5041 tgagcgggct tttttttaat tcccctattt gtttattttt ctaaatacat tcaaatatgt
     5101 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5161 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5221 tttttgctca cccagaaacg ctcgtgaaag taaaagacgc agaggaccaa ttgggggcac
     5281 gagtgggata catagaactg gacttgaata gcggtaaaat ccttgagagt tttcgccctg
     5341 aagagcgttt tccaatgatg agcactttca aagttctgct atgtggagca gtattatccc
     5401 gtgtagatgc ggggcaagag caactcggac gacgaataca ctattcgcag aatgacttgg
     5461 ttgaatactc cccagtgaca gaaaagcacc ttacggacgg aatgacggta agagaattat
     5521 gtagtgccgc cataacgatg agtgataaca ctgcggcgaa cttacttctg acaaccatcg
     5581 gtggaccgaa ggaattaacc gcttttttgc acaatatggg agaccatgta actcgccttg
     5641 accgttggga accagaactg aatgaagcca taccaaacga cgagcgagac accacaatgc
     5701 ctgcggcaat ggcaacaaca ttacgcaaac tattaactgg cgaactactt actctggctt
     5761 cacggcaaca attaatagac tggcttgaag cggataaagt tgcaggacca ctactgcgtt
     5821 cggcacttcc tgctggctgg tttattgctg ataaatctgg ggcaggagag cgtggttcac
     5881 ggggtatcat tgccgcactt ggaccagatg gtaagccttc ccgtatcgta gttatctaca
     5941 cgacgggtag tcaggcaact atggacgaac gaaatagaca gattgctgaa ataggggctt
     6001 cactgattaa gcattggtaa accgatacaa ttaaaggctc cttttggagc cttttttttt
     6061 ggacggaccg gtagaaaaga tcaaaggatc ttc
//
