LOCUS       sequence-127243-        6462 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 6462)
  AUTHORS   Shibata M, Lu T, Furuya T, Degterev A, Mizushima N, Yoshimori T, 
            MacDonald M, Yankner B, Yuan J
  TITLE     Regulation of intracellular accumulation of mutant Huntingtin by 
            Beclin 1.
  JOURNAL   J Biol Chem. 2006 May 19. 281(20):14474-85.
  PUBMED    16522639
REFERENCE   2  (bases 1 to 6462)
  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: "J Biol 
            Chem. 2006 May 19. 281(20):14474-85."
COMMENT     Sequence Label: pcDNA3-Beclin1
FEATURES             Location/Qualifiers
     source          1..6462
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(108..696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(188..207)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    1025..1072
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1097..1116
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1304..2098)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(1304..2098)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1415..1434)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2025..2044
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2133..2490)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2147..2282
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        2299..2490
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2447..2467
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(2550..3341,3342..3410))
                     /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     3173..3192
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3411..3515)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3542..3997
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3629..3648)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3839..3860
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     enhancer        4009..4388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4389..4592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4543..4563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4637..4656
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4637..4655
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4748..4771
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(6208..6225)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6214..6438
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
       61 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
      121 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      181 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
      241 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
      301 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
      361 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      421 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
      481 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
      541 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
      601 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
      661 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
      721 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
      781 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
      841 atatatgagt aacctgaggc tatggcaggg cctgccgccc cgacgttggc tgcgagccct
      901 gggccttcac ccgaacttgg ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg
      961 ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt
     1021 ttatgaacaa acgacccaac accgtgcgtt ttattctgtc tttttattgc cgtcatagcg
     1081 cgggttcctt ccggtattgt ctccttccgt gtttcagtta gcctccccct agggtgggcg
     1141 aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg
     1201 attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg
     1261 ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa
     1321 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     1381 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     1441 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     1501 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
     1561 cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     1621 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
     1681 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
     1741 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
     1801 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
     1861 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca
     1921 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
     1981 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
     2041 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc
     2101 gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa agcctaggcc tccaaaaaag
     2161 cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata
     2221 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
     2281 atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata
     2341 cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc
     2401 atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac
     2461 acattccaca gctggttctt tccgcctcag gactcttcct ttttcaataa atcaatctaa
     2521 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2581 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2641 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     2701 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2761 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     2821 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     2881 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     2941 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3001 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3061 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3121 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3181 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3241 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3301 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3361 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3421 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3481 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3541 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3601 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3661 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3721 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3781 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3841 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3901 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3961 aacgcgaatt ttaacaaaat attaacgctt acaatttacg cgcgcgttga cattgattat
     4021 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     4081 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     4141 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     4201 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     4261 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     4321 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     4381 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     4441 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     4501 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     4561 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt
     4621 actggcttat cgaaattaat acgactcact atagggagac ccaagctggc tagcgtttaa
     4681 acttaagctt ggtaccgagc tcggatccac tagtccagtg tggtggaatt cggctttagc
     4741 tagcatggac tacaaggacg atgatgacaa aatggaaggg tctaagacgt ccaacaacag
     4801 caccatgcag gtgagcttcg tgtgccagcg ctgcagccag cccctgaaac tggacacgag
     4861 tttcaagatc ctggaccgtg tcaccatcca ggaactcaca gctccattac ttaccacagc
     4921 ccaggcgaaa ccaggagaga cccaggagga agagactaac tcaggagagg agccatttat
     4981 tgaaactcct cgccaggatg gtgtctctcg cagattcatc cccccagcca ggatgatgtc
     5041 cacagaaagt gccaacagct tcactctgat tggggaggta tctgatggcg gcaccatgga
     5101 gaacctcagc cgaagactga aggtcactgg ggaccttttt gacatcatgt cgggccagac
     5161 agatgtggat cacccactct gtgaggaatg cacagatact cttttagacc agctggacac
     5221 tcagctcaac gtcactgaaa atgagtgtca gaactacaaa cgctgtttgg agatcttaga
     5281 gcaaatgaat gaggatgaca gtgaacagtt acagatggag ctaaaggagc tggcactaga
     5341 ggaggagagg ctgatccagg agctggaaga cgtggaaaag aaccgcaaga tagtggcaga
     5401 aaatctcgag aaggtccagg ctgaggctga gagactggat caggaggaag ctcagtatca
     5461 gagagaatac agtgaattta aacgacagca gctggagctg gatgatgagc tgaagagtgt
     5521 tgaaaaccag atgcgttatg cccagacgca gctggataag ctgaagaaaa ccaacgtctt
     5581 taatgcaacc ttccacatct ggcacagtgg acagtttggc acaatcaata acttcaggct
     5641 gggtcgcctg cccagtgttc ccgtggaatg gaatgagatt aatgctgctt ggggccagac
     5701 tgtgttgctg ctccatgctc tggccaataa gatgggtctg aaatttcaga gataccgact
     5761 tgttccttac ggaaaccatt catatctgga gtctctgaca gacaaatcta aggagctgcc
     5821 gttatactgt tctggggggt tgcggttttt ctgggacaac aagtttgacc atgcaatggt
     5881 ggctttcctg gactgtgtgc agcagttcaa agaagaggtt gagaaaggcg agacacgttt
     5941 ttgtcttccc tacaggatgg atgtggagaa aggcaagatt gaagacacag gaggcagtgg
     6001 cggctcctat tccatcaaaa cccagtttaa ctctgaggag cagtggacaa aagctctcaa
     6061 gttcatgctg acgaatctta agtggggtct tgcttgggtg tcctcacaat tttataacaa
     6121 atgagctagc taaagccgaa ttctgcagat atccagcaca gtggcggccg ctcgagtcta
     6181 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     6241 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     6301 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     6361 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     6421 atgcggtggg ctctatggct tctgaggcgg aaagaaccag tg
//
