LOCUS       sequence-172731-        7822 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7822)
  AUTHORS   Sun Q, Fan W, Chen K, Ding X, Chen S, Zhong Q
  TITLE     Identification of Barkor as a mammalian autophagy-specific factor 
            for Beclin 1 and class III phosphatidylinositol 3-kinase.
  JOURNAL   Proc Natl Acad Sci U S A. 2008 Dec 9. 105(49):19211-6.
  PUBMED    19050071
REFERENCE   2  (bases 1 to 7822)
  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: "Proc Natl 
            Acad Sci U S A. 2008 Dec 9. 105(49):19211-6."
COMMENT     Sequence Label: pcDNA4-Vps34-Flag
FEATURES             Location/Qualifiers
     source          1..7822
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1048..1057
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3724..3789
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3830..3847)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3836..4060
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4106..4534
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4193..4212)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4403..4424
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4543..4563)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4548..4877
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4728..4863
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4790..4809
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4925..4972
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4991..5365
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5495..5616
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5532..5551)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5586..5605
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5665..5681)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5665..5681)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5678..5700)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5689..5705
                     /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(5713..5719,5720..5737,5738..5743))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5758..5779
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5896..5913)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6067..6655)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6147..6166)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6826..7617,7618..7686))
                     /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     7449..7468
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7687..7791)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga
     1021 attctgcaga tatccagcac agtggcggcc gccatggggg aagcagagaa gtttcactac
     1081 atctatagtt gtgacctgga tatcaacgtc cagcttaaga taggaagctt ggaagggaag
     1141 agagaacaaa agagttataa agctgtcctg gaagacccaa tgttgaagtt ctcaggacta
     1201 tatcaagaga catgctctga tctttatgtt acttgtcaag tttttgcaga agggaagcct
     1261 ttggccttgc cagtgagaac atcctacaaa gcatttagta caagatggaa ctggaatgaa
     1321 tggctgaaac taccagtaaa ataccctgac ctgcccagga atgcccaagt ggccctcacc
     1381 atatgggatg tgtatggtcc cggaaaagca gtgcctgtag gaggaacaac ggtttcgctc
     1441 tttggaaaat acggcatgtt tcgccaaggg atgcatgact tgaaagtctg gcctaatgta
     1501 gaagcagatg gatcagaacc cacaaaaact cctggcagaa caagtagcac tctctcagaa
     1561 gatcagatga gccgtcttgc caagctcacc aaagctcatc gacaaggaca catggtgaaa
     1621 gtagattggc tggatagatt gacatttaga gaaatagaaa tgataaatga gagtgaaaaa
     1681 cgaagttcta atttcatgta cctgatggtt gaatttcgat gtgtcaagtg tgatgataag
     1741 gaatatggta ttgtttatta tgaaaaggac ggtgatgaat catctccaat tttaacaagt
     1801 tttgaattag tgaaagttcc tgacccccag atgtctatgg agaatttagt tgagagcaaa
     1861 caccacaagc ttgcccggag tttaagaagt ggaccttctg accacgatct gaaacccaat
     1921 gctgccacga gagatcagtt aaatattatt gtgagttatc caccaaccaa gcaacttaca
     1981 tatgaagaac aagatcttgt ttggaagttt agatattatc ttacgaatca agaaaaagcc
     2041 ttgacaaaat tcttgaaatg tgttaattgg gatctacctc aagaggccaa acaggccttg
     2101 gaacttctgg gaaaatggaa gccgatggat gtagaggact ccttggagct gttatcctct
     2161 cattacacca acccaactgt gaggcgttat gctgttgccc ggttgcgaca ggccgatgat
     2221 gaggatttgt tgatgtacct attacaattg gtccaggctc tcaaatatga aaattttgat
     2281 gatataaaga atggattgga acctaccaag aaggatagtc agagttcagt gtcagaaaat
     2341 gtgtcaaatt ctggaataaa ttctgcagaa atagatagct cccaaattat aaccagcccc
     2401 cttccttcag tctcttcacc tcctcctgca tcaaaaacaa aagaagttcc agatggcgaa
     2461 aatctggaac aagatctctg taccttcttg atatcgagag cctgcaaaaa ctcaacactg
     2521 gctaattatt tatactggta tgtgatagtg gaatgtgaag atcaagatac tcagcagaga
     2581 gatccaaaga cccatgagat gtacttgaac gtaatgagaa gattcagcca agcattgttg
     2641 aagggtgata agtctgtcag agttatgcgt tctttgctgg ctgcacaaca gacatttgta
     2701 gatcggttgg tgcatctaat gaaggcagta caacgcgaaa gtggaaatcg taagaaaaag
     2761 aatgagagac tacaggcatt gcttggagat aatgaaaaga tgaatttgtc agatgtggaa
     2821 cttatcccgt tgcctttaga accccaagtg aaaattagag gaataattcc ggaaacagct
     2881 acactgttta aaagtgccct tatgcctgca cagttgtttt ttaagacgga agatggaggc
     2941 aaatatccag ttatatttaa gcatggagat gatttacgtc aagatcaact tattcttcaa
     3001 atcatttcac tcatggacaa gctgttacgg aaagaaaatc tggacttgaa attgacacct
     3061 tataaggtgt tagccaccag tacaaaacat ggcttcatgc agtttatcca gtcagttcct
     3121 gtggctgaag ttcttgatac agagggaagc attcagaact tttttagaaa atatgcacca
     3181 agtgagaatg ggccaaatgg gattagtgct gaggtcatgg acacttacgt taaaagctgt
     3241 gctggatatt gcgtgatcac ctatatactt ggagttggag acaggcacct ggataacctt
     3301 ttgctaacaa aaacaggcaa actcttccac atagactttg gatatatttt gggtcgggat
     3361 ccaaagcctc ttcctccacc aatgaagctg aataaagaaa tggtagaagg aatggggggc
     3421 acacagagtg agcagtacca agagttccgt aaacagtgtt acacggcttt cctccacctg
     3481 cgaaggtatt ctaatctgat tttgaacttg ttttccttga tggttgatgc aaacattcca
     3541 gatattgcac ttgaaccaga taaaactgtg aaaaaggttc aggataaatt ccgcttagac
     3601 ctgtcggatg aagaggctgt gcattacatg cagagtctga ttgatgagag tgtccatgct
     3661 ctttttgctg cagtggtgga acagattcac aagtttgccc agtactggag aaaaggcggc
     3721 cgcgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     3781 gatgacaagt gactcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     3841 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3901 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3961 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     4021 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     4081 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4141 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4201 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4261 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4321 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4381 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4441 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4501 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4561 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4621 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4681 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4741 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4801 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4861 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
     4921 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     4981 gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc
     5041 cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga
     5101 cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt
     5161 ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga
     5221 gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat
     5281 cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca
     5341 cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt
     5401 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5461 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5521 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5581 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5641 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5701 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5761 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5821 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5881 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5941 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6001 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6061 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6121 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6181 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6241 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6301 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6361 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6421 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6481 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6541 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6601 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6661 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6721 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6781 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6841 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6901 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6961 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7021 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7081 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7141 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7201 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7261 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7321 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7381 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7441 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7501 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7561 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7621 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7681 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7741 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7801 ccgaaaagtg ccacctgacg tc
//
