LOCUS       sequence-169128-        6459 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 6459)
  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 6459)
  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-Beclin1 (FL)
FEATURES             Location/Qualifiers
     source          1..6459
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        143..522
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        523..726
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     677..697
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    728..746
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    749..767
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     777..801
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2269..2334
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2375..2392)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2381..2605
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2651..3079
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2738..2757)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2948..2969
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3088..3108)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3093..3422
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3273..3408
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3335..3354
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3470..3517
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3536..3910
                     /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    4040..4161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4077..4096)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4131..4150
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4210..4226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4210..4226)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4223..4245)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4234..4250
                     /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(4258..4264,4265..4282,4283..4288))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4303..4324
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4441..4458)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4612..5200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4692..4711)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5371..6162,6163..6231))
                     /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     5994..6013
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6232..6336)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6411..6430)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
       61 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
      121 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      181 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      241 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      301 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      361 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      421 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      481 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      541 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      601 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      661 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      721 agagctctcc ctatcagtga tagagatctc cctatcagtg atagagatcg tcgacgagct
      781 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
      841 agacaccggg accgatccag cctccggact ctagcgttta aacttaagct tggtaccgag
      901 ctcggatcca tggaagggtc taagacgtcc aacaacagca ccatgcaggt gagcttcgtg
      961 tgccagcgct gcagccagcc cctgaaactg gacacgagtt tcaagatcct ggaccgtgtc
     1021 accatccagg aactcacagc tccattactt accacagccc aggcgaaacc aggagagacc
     1081 caggaggaag agactaactc aggagaggag ccatttattg aaactcctcg ccaggatggt
     1141 gtctctcgca gattcatccc cccagccagg atgatgtcca cagaaagtgc caacagcttc
     1201 actctgattg gggaggcatc tgatggcggc accatggaga acctcagccg aagactgaag
     1261 gtcactgggg acctttttga catcatgtcg ggccagacag atgtggatca cccactctgt
     1321 gaggaatgca cagatactct tttagaccag ctggacactc agctcaacgt cactgaaaat
     1381 gagtgtcaga actacaaacg ctgtttggag atcttagagc aaatgaatga ggatgacagt
     1441 gaacagttac agatggagct aaaggagctg gcactagagg aggagaggct gatccaggag
     1501 ctggaagacg tggaaaagaa ccgcaagata gtggcagaaa atctcgagaa ggtccaggct
     1561 gaggctgaga gactggatca ggaggaagct cagtatcaga gagaatacag tgaatttaaa
     1621 cgacagcagc tggagctgga tgatgagctg aagagtgttg aaaaccagat gcgttatgcc
     1681 cagacgcagc tggataagct gaagaaaacc aacgtcttta atgcaacctt ccacatctgg
     1741 cacagtggac agtttggcac aatcaataac ttcaggctgg gtcgcctgcc cagtgttccc
     1801 gtggaatgga atgagattaa tgctgcttgg ggccagactg tgttgctgct ccatgctctg
     1861 gccaataaga tgggtctgaa atttcagaga taccgacttg ttccttacgg aaaccattca
     1921 tatctggagt ctctgacaga caaatctaag gagctgccgt tatactgttc tggggggttg
     1981 cggtttttct gggacaacaa gtttgaccat gcaatggtgg ctttcctgga ctgtgtgcag
     2041 cagttcaaag aagaggttga gaaaggcgag acacgttttt gtcttcccta caggatggat
     2101 gtggagaaag gcaagattga agacacagga ggcagtggcg gctcctattc catcaaaacc
     2161 cagtttaact ctgaggagca gtggacaaaa gctctcaagt tcatgctgac gaatcttaag
     2221 tggggtcttg cttgggtgtc ctcacaattt tataacaaag gcggccgcga ctacaaagac
     2281 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtgactc
     2341 gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     2401 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2461 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     2521 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     2581 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     2641 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2701 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2761 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2821 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2881 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2941 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3001 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3061 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3121 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3181 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3241 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3301 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3361 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3421 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
     3481 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc
     3541 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt
     3601 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt
     3661 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac
     3721 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt
     3781 gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg
     3841 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga
     3901 gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     3961 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4021 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4081 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4141 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     4201 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4261 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4321 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4381 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4441 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4501 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4561 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4621 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4681 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4741 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4801 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4861 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4921 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4981 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5041 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5101 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5161 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5221 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5281 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5341 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5401 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5461 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5521 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5581 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5641 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5701 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5761 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5821 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5881 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5941 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6001 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6061 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6121 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6181 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6241 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6301 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6361 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     6421 tctgatgccg catagttaag ccagtatctg ctccctgct
//
