LOCUS       sequence-472454-        6300 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of FL human Beclin 1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6300)
  AUTHORS   Ohashi Y, Tremel S, Masson GR, McGinney L, Boulanger J, 
            Rostislavleva K, Johnson CM, Niewczas I, Clark J, Williams RL
  TITLE     Membrane characteristics tune activities of endosomal and autophagic
            human VPS34 complexes.
  JOURNAL   Elife. 2020 Jun 30;9:e58281. doi: 10.7554/eLife.58281.
  PUBMED    32602837
REFERENCE   2  (bases 1 to 6300)
  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: "2020-06-30"; volume: "9:e58281. doi"; pages: " 
            10.7554/eLife.58281"
COMMENT     Sequence Label: pYO1006
FEATURES             Location/Qualifiers
     source          1..6300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          784..1800
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1724..1746
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1808..1827
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1874..1883
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3322..3341)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3387..3442
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3441..3460)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3803..3819)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3803..3819)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3816..3838)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3827..3843
                     /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(3851..3857,3858..3875,3876..3881))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3896..3917
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3976..4171
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4022..4157
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4177..4311
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4214..4233)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4268..4287
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4379..4396)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4550..5138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4630..4649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5309..6100,6101..6169))
                     /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     5932..5951
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6170..6274)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggtgg gggggggggg gggggggggg gggggggggg
      541 gggggggggg gggggggggg gggggggggg cgggggccag gggggggggg gggctcaaaa
      601 tgttctttac tatgccgggg ggggggggcg gcgggggggg ggggggggcg gggggggggg
      661 gggggggggg cgagggggag aggtgcggcg gcagccaatc ggagcggcgc gctccgaaag
      721 tttcctttta gggcggggcg gcggcggcgg cggccctata aaaagcgagg cgcgcggcgg
      781 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
      841 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
      901 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
      961 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg
     1021 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag
     1081 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc
     1141 gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt
     1201 gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg
     1261 cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg
     1321 cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg
     1381 gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc
     1441 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
     1501 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
     1561 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
     1621 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt
     1681 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
     1741 gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag
     1801 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attgctcgat
     1861 cgacggtatc gatgccacca tggaagggtc taagacgtcc aacaacagca ccatgcaggt
     1921 gagcttcgtg tgccagcgct gcagccagcc cctgaaactg gacacgagtt tcaagatcct
     1981 ggaccgtgtc accatccagg aactcacagc tccattactt accacagccc aggcgaaacc
     2041 aggagagacc caggaggaag agactaactc aggagaggag ccatttattg aaactcctcg
     2101 ccaggatggt gtctctcgca gattcatccc cccagccagg atgatgtcca cagaaagtgc
     2161 caacagcttc actctgattg gggaggcatc tgatggcggc accatggaga acctcagccg
     2221 aagactgaag gtcactgggg acctttttga catcatgtcg ggccagacag atgtggatca
     2281 cccactctgt gaggaatgca cagatactct tttagaccag ctggacactc agctcaacgt
     2341 cactgaaaat gagtgtcaga actacaaacg ctgtttggag atcttagagc aaatgaatga
     2401 ggatgacagt gaacagttac agatggagct aaaggagctg gcactagagg aggagaggct
     2461 gatccaggag ctggaagacg tggaaaagaa ccgcaagata gtggcagaaa atctcgagaa
     2521 ggtccaggct gaggctgaga gactggatca ggaggaagct cagtatcaga gagaatacag
     2581 tgaatttaaa cgacagcagc tggagctgga tgatgagctg aagagtgttg aaaaccagat
     2641 gcgttatgcc cagacgcagc tggataagct gaagaaaacc aacgtcttta atgcaacctt
     2701 ccacatctgg cacagtggac agtttggcac aatcaataac ttcaggctgg gtcgcctgcc
     2761 cagtgttccc gtggaatgga atgagattaa tgctgcttgg ggccagactg tgttgctgct
     2821 ccatgctctg gccaataaga tgggtctgaa atttcagaga taccgacttg ttccttacgg
     2881 aaaccattca tatctggagt ctctgacaga caaatctaag gagctgccgt tatactgttc
     2941 tggggggttg cggtttttct gggacaacaa gtttgaccat gcaatggtgg ctttcctgga
     3001 ctgtgtgcag cagttcaaag aagaggttga gaaaggcgag acacgttttt gtcttcccta
     3061 caggatggat gtggagaaag gcaagattga agacacagga ggcagtggcg gctcctattc
     3121 catcaaaacc cagtttaact ctgaggagca gtggacaaaa gctctcaagt tcatgctgac
     3181 gaatctgaag tggggtctgg cttgggtgtc ctctcaattt tataacaaat gactcgagga
     3241 attcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct
     3301 ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa
     3361 gccccttgag catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg
     3421 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca
     3481 tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa
     3541 caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt
     3601 attccataga aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt
     3661 tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct
     3721 ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacctgca
     3781 gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3841 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3901 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3961 cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4021 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4081 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4141 aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat
     4201 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4261 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt
     4321 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4381 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4441 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4501 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4561 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4621 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4681 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4741 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4801 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4861 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4921 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4981 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5041 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5101 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5161 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5221 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5281 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5341 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5401 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5461 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5521 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5581 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5641 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5701 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5761 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5821 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5881 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5941 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6001 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6061 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6121 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6181 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6241 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
