LOCUS       sequence-156195-        5388 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 5388)
  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 5388)
  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 151-241
FEATURES             Location/Qualifiers
     source          1..5388
                     /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"
     CDS             1056..1121
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1162..1179)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1168..1392
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1438..1866
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1525..1544)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1735..1756
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1875..1895)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1880..2209
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2060..2195
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2122..2141
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2257..2304
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2323..2697
                     /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    2827..2948
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2864..2883)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2918..2937
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2997..3013)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2997..3013)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3010..3032)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3021..3037
                     /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(3045..3051,3052..3069,3070..3075))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3090..3111
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3228..3245)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3399..3987)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3479..3498)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4158..4949,4950..5018))
                     /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     4781..4800
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5019..5123)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5198..5217)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
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 gtaccgagct cggatccatg cagctcaacg
      781 tcactgaaaa tgagtgtcag aactacaaac gctgtttgga gatcttagag caaatgaatg
      841 aggatgacag tgaacagtta cagatggagc taaaggagct ggcactagag gaggagaggc
      901 tgatccagga gctggaagac gtggaaaaga accgcaagat agtggcagaa aatctcgaga
      961 aggtccaggc tgaggctgag agactggatc aggaggaagc tcagtatcag agagaataca
     1021 gtgaatttaa acgacagcag ctggagggcg gccgcgacta caaagaccat gacggtgatt
     1081 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtgactcgag tctagagggc
     1141 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     1201 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     1261 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     1321 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     1381 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     1441 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1501 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     1561 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1621 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     1681 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     1741 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     1801 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     1861 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     1921 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     1981 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     2041 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     2101 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     2161 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     2221 agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag
     2281 tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt
     2341 gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg
     2401 ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg
     2461 accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg
     2521 tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc
     2581 cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc
     2641 gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac
     2701 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     2761 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     2821 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2881 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2941 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     3001 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     3061 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     3121 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     3181 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3241 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3301 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3361 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3421 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3481 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3541 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3601 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3661 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3721 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3781 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3841 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3901 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3961 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4021 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4081 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4141 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4201 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4261 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4321 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4381 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4441 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4501 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4561 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4621 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4681 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4741 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4801 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4861 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4921 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4981 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5041 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5101 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     5161 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
     5221 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     5281 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
     5341 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgtt
//
