LOCUS       sequence-472457-        9300 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of FL human Beclin 1 and FL human UVRAG in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9300)
  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 9300)
  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: pYO1031
FEATURES             Location/Qualifiers
     source          1..9300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    586..604
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    607..625
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     635..659
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(2146..2163)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2152..2376
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        2433..2736
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          3036..4052
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3976..3998
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4060..4079
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(6321..6340)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6386..6441
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6440..6459)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6802..6818)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6802..6818)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6815..6837)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6826..6842
                     /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(6850..6856,6857..6874,6875..6880))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6895..6916
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6975..7170
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7021..7156
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    7176..7310
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7213..7232)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7267..7286
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7378..7395)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7549..8137)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7629..7648)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8308..9099,9100..9168))
                     /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     8931..8950
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9169..9273)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata
      601 gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg
      661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      721 tccggactct agcgtttaaa cttaagcttg gtaccatgga agggtctaag acgtccaaca
      781 acagcaccat gcaggtgagc ttcgtgtgcc agcgctgcag ccagcccctg aaactggaca
      841 cgagtttcaa gatcctggac cgtgtcacca tccaggaact cacagctcca ttacttacca
      901 cagcccaggc gaaaccagga gagacccagg aggaagagac taactcagga gaggagccat
      961 ttattgaaac tcctcgccag gatggtgtct ctcgcagatt catcccccca gccaggatga
     1021 tgtccacaga aagtgccaac agcttcactc tgattgggga ggcatctgat ggcggcacca
     1081 tggagaacct cagccgaaga ctgaaggtca ctggggacct ttttgacatc atgtcgggcc
     1141 agacagatgt ggatcaccca ctctgtgagg aatgcacaga tactctttta gaccagctgg
     1201 acactcagct caacgtcact gaaaatgagt gtcagaacta caaacgctgt ttggagatct
     1261 tagagcaaat gaatgaggat gacagtgaac agttacagat ggagctaaag gagctggcac
     1321 tagaggagga gaggctgatc caggagctgg aagacgtgga aaagaaccgc aagatagtgg
     1381 cagaaaatct cgagaaggtc caggctgagg ctgagagact ggatcaggag gaagctcagt
     1441 atcagagaga atacagtgaa tttaaacgac agcagctgga gctggatgat gagctgaaga
     1501 gtgttgaaaa ccagatgcgt tatgcccaga cgcagctgga taagctgaag aaaaccaacg
     1561 tctttaatgc aaccttccac atctggcaca gtggacagtt tggcacaatc aataacttca
     1621 ggctgggtcg cctgcccagt gttcccgtgg aatggaatga gattaatgct gcttggggcc
     1681 agactgtgtt gctgctccat gctctggcca ataagatggg tctgaaattt cagagatacc
     1741 gacttgttcc ttacggaaac cattcatatc tggagtctct gacagacaaa tctaaggagc
     1801 tgccgttata ctgttctggg gggttgcggt ttttctggga caacaagttt gaccatgcaa
     1861 tggtggcttt cctggactgt gtgcagcagt tcaaagaaga ggttgagaaa ggcgagacac
     1921 gtttttgtct tccctacagg atggatgtgg agaaaggcaa gattgaagac acaggaggca
     1981 gtggcggctc ctattccatc aaaacccagt ttaactctga ggagcagtgg acaaaagctc
     2041 tcaagttcat gctgacgaat ctgaagtggg gtctggcttg ggtgtcctct caattttata
     2101 acaaatgagt cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt
     2161 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     2221 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     2281 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     2341 atagcaggca tgctggggat gcggtgggct ctatggtatt aatagtaatc aattacgggg
     2401 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     2461 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     2521 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     2581 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     2641 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     2701 cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc
     2761 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
     2821 ttattttgtg cagcgatggg ggcggggggg gggggggggg gggcccccgg gcgggggggg
     2881 gggggggggg gggggggggg gggggggggg gggggggcgg ggcgaggcgg agaggtgcgg
     2941 cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc
     3001 ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc
     3061 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
     3121 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt
     3181 aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc
     3241 ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg
     3301 cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg
     3361 cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg
     3421 gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg
     3481 tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg
     3541 gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg
     3601 gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct
     3661 cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca
     3721 gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct
     3781 gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc
     3841 ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc
     3901 gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg
     3961 gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta
     4021 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc
     4081 tgtctcatca ttttggcaaa gaattgctcg atcgacggta tcgatgccac catgagcgcc
     4141 tccgcgtcgg tcgggggcca ggtcccccag ccacccccgg gcccggccgc tgctctgcct
     4201 cccggttctg ccgcgcgggc cctgcatgtg gagctgccgt ctcagcagcg gcgtcttcga
     4261 catcttcgga acattgctgc ccggaacatt gttaatagaa atggccatca gctccttgat
     4321 acctacttta cacttcactt gtgtagtact gaaaagatat ataaagaatt ttatagaagt
     4381 gaagtgatta agaattcctt gaatcccacg tggcgaagtc tcgattttgg aattatgcca
     4441 gaccgtcttg atacatctgt gtcttgtttc gtggtgaaga tatggggtgg aaaggagaac
     4501 atctaccagc tgttgattga atggaaagtc tgtttggatg ggctgaaata cttgggtcag
     4561 cagattcatg cccgaaacca aaatgaaata atttttgggc tgaatgatgg atactatggt
     4621 gctccatttg aacataaggg ttattcaaat gctcagaaga ctattcttct gcaggtggat
     4681 cagaactgtg ttcgcaattc ttacgatgtc ttctctttgc tacggcttca tagagcccag
     4741 tgtgcaatta aacagactca ggtaactgtt cagaaaattg gaaaggaaat tgaagaaaaa
     4801 ctaagactca catctacaag caatgaactg aaaaaaaaaa gtgaatgcct gcagttaaaa
     4861 attttggtgc ttcagaatga actggaacgg cagaagaaag ctttgggacg ggaggtggca
     4921 ttactgcata agcaacaaat tgcattacaa gacaaaggaa gtgcattttc agctgagcac
     4981 ctcaaacttc aactccagaa ggaatcccta aatgagctga ggaaggagtg cactgcaaaa
     5041 agagaactct tcttgaagac taatgctcag ttgacaattc gttgcaggca gttactctct
     5101 gagctttcct acatttaccc tattgatttg aatgaacata aggattactt tgtatgcggt
     5161 gtcaagttgc ctaattctga ggacttccaa gcaaaagatg atggaagcat tgctgttgcc
     5221 cttggttata ctgcacatct ggtctccatg atttcctttt tcctacaagt gcccctcaga
     5281 tatcctataa ttcataaggg gtctagatca acaatcaaag acaatatcaa tgacaaactg
     5341 acggaaaagg agagagagtt tccactgtat ccaaaaggag gggagaagtt gcagtttgat
     5401 tatggtgtct atcttctgaa caaaaatata gcacagctaa gatatcaaca tggactaggg
     5461 actccagact tgcggcaaac ccttcccaac ctgaaaaact tcatggagca tggactaatg
     5521 gtcaggtgtg acagacatca cacctccagt gcaatccctg ttcctaagag acaaagctcc
     5581 atatttgggg gtgcagatgt aggcttctct ggggggatcc cttcaccaga caaaggacat
     5641 cgaaaacggg ccagctctga gaatgagaga cttcagtaca aaacccctcc tcccagttac
     5701 aactcagcat tagcccagcc tgtgaccacc gtcccctcca tgggagagac cgagagaaag
     5761 ataacatctc tatcctcctc cttggatacc tccttggact tctccaaaga aaacaagaaa
     5821 aaaggagagg atctagttgg cagcttaaac ggaggccacg cgaatgtgca ccctagccaa
     5881 gaacaaggag aagccctctc cgggcaccgg gccacagtca atggcactct cctacccagc
     5941 gagcaggccg ggtccgccag tgtccagctt ccaggcgagt tccacccagt ctcagaagct
     6001 gagctctgct gtactgtgga gcaagcagaa gaaatcatcg ggctggaagc cacaggtttc
     6061 gcctcaggtg atcagctaga agcatttaac tgcatcccag tggacagtgc tgtggcagta
     6121 gagtgtgacg aacaagttct gggagaattt gaagagttct cccgaaggat ctatgcactg
     6181 aatgaaaacg tatccagctt ccgccggccg cgcaggagtt ccgataagta actcgaggaa
     6241 ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg
     6301 gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag
     6361 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     6421 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
     6481 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
     6541 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
     6601 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
     6661 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
     6721 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag
     6781 cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6841 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6901 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6961 gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     7021 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     7081 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     7141 ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg
     7201 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     7261 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta
     7321 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     7381 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     7441 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     7501 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     7561 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7621 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7681 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7741 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7801 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7861 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7921 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7981 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     8041 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     8101 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     8161 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     8221 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     8281 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     8341 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     8401 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     8461 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     8521 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     8581 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     8641 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8701 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8761 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8821 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8881 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8941 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     9001 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     9061 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     9121 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     9181 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     9241 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgg
//
