LOCUS       sequence-337507-        6221 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  For the purifcation of WIPI2d protein .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6221)
  AUTHORS   Chang C, Shi X, Jensen LE, Yokom AL, Fracchiolla D, Martens S, 
            Hurley JH
  TITLE     Reconstitution of cargo-induced LC3 lipidation in mammalian 
            selective autophagy.
  JOURNAL   Sci Adv. 2021 Apr 23;7(17). pii: 7/17/eabg4922. doi: 
            10.1126/sciadv.abg4922. Print 2021 Apr.
  PUBMED    33893090
REFERENCE   2  (bases 1 to 6221)
  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: "Sci Adv."; 
            date: "2021-04-23"; volume: "7(17). pii: 7/17/eabg4922. doi"; pages:
            " 10.1126/sciadv.abg4922. Print 2021 Apr"
COMMENT     Sequence Label: WIPI2d-TEVcs-TSF
FEATURES             Location/Qualifiers
     source          1..6221
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     enhancer        13..392
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          637..1653
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1577..1599
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1661..1680
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3014..3034
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     regulatory      3039..3048
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3050..3073
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3110..3133
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3251..3270)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3316..3371
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3370..3389)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3732..3748)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3732..3748)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3745..3767)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3756..3772
                     /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)."
     promoter        complement(join(3780..3786,3787..3804,3805..3810))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3825..3846
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3905..4100
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3951..4086
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4106..4240
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4143..4162)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4197..4216
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4308..4325)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4479..5067)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4559..4578)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5238..6029,6030..6098))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5861..5880
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6099..6203)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gccacctggg tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
       61 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      121 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      181 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      241 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      301 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      361 acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca
      421 ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat
      481 tttgtgcagc gatgggggcg gggggggggg gggcggggcg gggcgaggcg gagaggtgcg
      541 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
      601 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
      661 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
      721 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
      781 taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc
      841 cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc
      901 gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt
      961 gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg
     1021 ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt
     1081 gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt tgctgagcac
     1141 ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc
     1201 ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggagggc
     1261 tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc ggcgagccgc
     1321 agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc
     1381 tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag
     1441 cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc
     1501 cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg
     1561 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct
     1621 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg
     1681 ctgtctcatc attttggcaa agaattcctc gatcgacggt atcgatgcca ccatgaacct
     1741 ggcgagccag agcggggagg ccggcgccgg ccagctgctc ttcgccaact tcaaccagga
     1801 caacacgtcc ctagctgttg gtagtaagtc cggttataaa tttttctccc tttcttctgt
     1861 ggataagctg gaacagatct atgaatgcac cgatacggaa gatgtgtgca ttgtagagag
     1921 attgttctcc agcagcctag tggccatcgt gagccttaaa gcaccaagga agctaaaggt
     1981 ttgccacttt aagaagggaa ctgagatctg caactacagc tactccaaca cgattctggc
     2041 tgtgaagctc aacaggcaga ggctgatagt atgcctggag gagtccctgt acatccacaa
     2101 cattcgggac atgaaggtgc tgcatacgat cagggagacg cctccaaacc ctgcaggcct
     2161 gtgtgcgctg tcaatcaaca acgacaactg ctacttggcg tacccaggga gcgcgaccat
     2221 cggagaggtg caggtcttcg ataccattaa tttgagagct gcaaacatga ttccggctca
     2281 cgacagtcct ttagcggcac tggcctttga cgcaagtgga actaaacttg ccacggcttc
     2341 ggagaagggg accgtgatta gggtattttc cattccagaa ggacaaaaac tctttgagtt
     2401 tcggagagga gtaaagaggt gcgtgagcat ctgctccctg gccttcagca tggacggcat
     2461 gttcctctcc gcctccagca acactgagac cgtgcacatc ttcaaactcg agactgtgaa
     2521 agaaaaaccc ccagaggagc ccaccacctg gaccgggtac ttcgggaaag tgctcatggc
     2581 ctccaccagc tacctgcctt cccaagtgac agaaatgttc aaccagggca gagccttcgc
     2641 cacggtccgc ctgccattct gcggccacaa aaacatctgc tcgctagcca caattcagaa
     2701 gatcccgcgg ttgttggtgg gtgccgccga cgggtacctg tacatgtaca acctggaccc
     2761 ccaggagggc ggcgagtgtg ccctgatgaa gcagcaccgg ctggacggca gtctggaaac
     2821 gaccaatgag atcttggact ctgcctctca cgactgcccc ttagtcactc agacatacgg
     2881 cgcagctgca ggaaaagcct acacagacga cctgggtgct gtgggtggcg cctgcctgga
     2941 ggacgaggcc agcgccctgc gcctggatga ggacagcgag cacccgccca tgattcttcg
     3001 gactgacgga tcggaaaacc tgtattttca gggtggtacc atggcggcct ggtcacaccc
     3061 gcagttcgag aaaggtggag gtgcccgagg tggatcggga ggtggatcgt ggagccaccc
     3121 gcagttcgaa aaatgacccg ggtgatatct taattaatta actcgaggaa ttcactcctc
     3181 aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat
     3241 accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
     3301 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     3361 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
     3421 atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc
     3481 tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa
     3541 aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa
     3601 catccctaaa attttcctta catgttttac tagccagatt tttcctcctc tcctgactac
     3661 tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag cccaagcttg
     3721 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3781 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3841 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg
     3901 atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3961 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4021 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4081 gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata
     4141 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4201 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc
     4261 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4321 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4381 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4441 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4501 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4561 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4621 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4681 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4741 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4801 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4861 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4921 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4981 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5041 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5101 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5161 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5221 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5281 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5341 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5401 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5461 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5521 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5581 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5641 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5701 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5761 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5821 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5881 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5941 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6001 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6061 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6121 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6181 aataaacaaa taggggttcc gcgcacattt ccccgaaaag t
//
