LOCUS       sequence-342321-        8631 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for expression of 3xFLAG-tagged truncated AMBRA1 that cannot 
            bind DDB1 (aa 1-34 deleted).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8631)
  AUTHORS   Chaikovsky AC, Li C, Jeng EE, Loebell S, Lee MC, Murray CW, Cheng R,
            Demeter J, Swaney DL, Chen SH, Newton BW, Johnson JR, Drainas AP, 
            Shue YT, Seoane JA, Srinivasan P, He A, Yoshida A, Hipkins SQ, 
            McCrea E, Poltorack CD, Krogan NJ, Diehl JA, Kong C, Jackson PK, 
            Curtis C, Petrov DA, Bassik MC, Winslow MM, Sage J
  TITLE     The AMBRA1 E3 ligase adaptor regulates the stability of cyclin D.
  JOURNAL   Nature. 2021 Apr;592(7856):794-798. doi: 10.1038/s41586-021-03474-7.
            Epub 2021 Apr 14.
  PUBMED    33854239
REFERENCE   2  (bases 1 to 8631)
  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"; doi: 
            "10.1038/s41586-021-03474-7"; journalName: "Nature"; date: 
            "2021-04"; volume: "592"; issue: "7856"; pages: "794-798"
COMMENT     Sequence Label: pcDNA4-deltaH-AMBRA1-3xFLAG
FEATURES             Location/Qualifiers
     source          1..8631
                     /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      990..999
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4533..4598
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(4639..4656)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4645..4869
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4915..5343
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5002..5021)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5212..5233
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5352..5372)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5357..5686
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5537..5672
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5599..5618
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5734..5781
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5800..6174
                     /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    6304..6425
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6341..6360)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6395..6414
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6474..6490)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6474..6490)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6487..6509)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6498..6514
                     /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(6522..6528,6529..6546,6547..6552))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6567..6588
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6705..6722)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6876..7464)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6956..6975)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7635..8426,8427..8495))
                     /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     8258..8277
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8496..8600)
                     /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 cttgatatcg ccaccatgga taaaacccgg tggatgaaat
     1021 gggagggcaa gagagtagaa ctgccggata gtccacgctc taccttctta ttggccttca
     1081 gcccagacag gactctctta gcctccaccc atgtgaacca taatatctat attacggagg
     1141 tgaagactgg caagtgtgtt cattccctga ttggacaccg ccgtactcca tggtgtgtca
     1201 cttttcatcc caccatctca ggccttattg cttctggctg cctagatggg gaggttagga
     1261 tttgggattt acacggtggc agtgaaagct ggttcacaga tagcaacaat gccattgcct
     1321 ccctggcttt ccaccctacg gctcagctcc tgctgattgc cactgccaat gagatccact
     1381 tctgggactg gagtcgacgg gaaccctttg ctgtggtgaa gacagctagt gagatggaac
     1441 gggtccgtct ggtgagattt gatccacttg gacactactt actcacagca attgttaacc
     1501 cctctaatca acagggtgat gacgaaccag agatccccat agatggaaca gaattatccc
     1561 actaccgtca gcgtgccctc ctgcaatcac agccagttcg ccggacgcct ctcctccaca
     1621 atttcctgca catgctgtcc tcccgctctt ctggcatcca gaccgagccc ttccatcccc
     1681 cggagcaggc ctcgtcaacg cagcaggacc agggcctcct gaaccggccg tctgccttca
     1741 gtacagtcca gagcagcact gccggcaaca cgctccgcaa cctcagtctg ggtcctaccc
     1801 gccgctcttt gggagggcct ctgtctagcc acccttctag gtatcaccga gaaatagctc
     1861 ctgggttgac aggatctgag tggacccgga cagtactcag tctgaactcc cgctctgagg
     1921 cggaatccat gcccccgccc agaaccagtg cctcttcggt gagtttgctg tctgtgctga
     1981 gacagcagga aggtggctct caggcatctg tgtacacttc agccacagaa gggaggggtt
     2041 ttccggcatc agggttggca actgagtcag atggagggaa tggctccagc caaaacaact
     2101 cgggcagcat tcgccatgag cttcagtgtg acctgagacg cttctttctg gagtatgacc
     2161 ggcttcagga gctggatcag agcctgagtg gggaagctcc ccagacccaa caggcccagg
     2221 aaatgctcaa caataacatt gaatctgaga ggccaggccc ttcccaccag cccaccccac
     2281 acagcagtga gaacaactcc aacctgtccc gtggccacct gaatcgctgt cgtgcttgcc
     2341 acaatctcct gaccttcaac aacgataccc tgcgctggga aagaaccaca cctaactact
     2401 cctctggcga ggctagttcc tcttggcagg tccccagctc ctttgagagt gtgccatcaa
     2461 gtggcagcca gttgccacct ctcgagcgga ctgagggcca aacgcccagc tccagcaggc
     2521 tggagttgag cagctctgct agtccgcagg aggagaggac tgtgggggtg gcctttaacc
     2581 aggagacagg ccactgggaa agaatttaca cccagtccag cagatctgga actgtgtcac
     2641 aggaggcctt acatcaggat atgcctgagg agagctctga ggaggattca ctcaggagga
     2701 ggctgctgga atcttccctc atttcattat cccgttatga tggagcagga tccagagagc
     2761 acccaattta cccagaccca gcgagattat ctcctgctgc atactacgcc cagaggatga
     2821 tccagtatct ctcacggaga gacagtattc gccagcgctc catgcgctac caacagaacc
     2881 gtctccgttc ttccacctcc tcctcttcct cagacaacca gggtccatca gtagagggaa
     2941 ccgacttgga atttgaggac tttgaggaca atggtgacag atccaggcac cgagctccac
     3001 gcaatgcccg gatgtctgca ccttcgcttg gacgctttgt cccaaggcgt ttcttgctgc
     3061 ctgagtactt gccttatgct gggatttttc atgaacgtgg acagcctggc ttggctactc
     3121 actcttctgt taacagggtc ctggcagggg cagtgatcgg tgatggacag tctgctgtgg
     3181 ccagtaacat tgccaatact acctaccggc tccagtggtg ggacttcact aagtttgacc
     3241 tccctgaaat cagtaatgct tccgtgaatg tgctggtgca gaactgcaag atctacaatg
     3301 atgccagctg tgacatttct gcagatggcc agctcctggc agctttcatc cccagcagcc
     3361 agaggggctt tcctgatgaa ggcatcctgg cagtgtactc cctggccccc cataacctgg
     3421 gcgaaatgct ctacaccaag cgatttggtc ccaatgccat ttcggtgagc ctgtccccaa
     3481 tgggcagata tgtaatggtg ggcttggcct cacgaaggat cctgctgcac ccctccacag
     3541 agcacatggt ggcccaggtc ttcaggctgc aacaggccca tggtggagag acctccatga
     3601 ggagagtttt caacgtcctt tatcccatgc ctgccgacca gcggagacat gtcagtatca
     3661 actctgcccg ttggctgcct gagccagggc ttggcttggc ctatggtact aacaaaggag
     3721 acctggtgat ctgccgacca gaggccttaa actctggtgt tgagtactac tgggaccagc
     3781 tgaacgagac ggtcttcact gtccattcca acagcaggag cagcgagcgg cctggaacca
     3841 gcagagccac atggaggaca gacagagaca tggggctgat gaatgccatt gggcttcagc
     3901 cccggaaccc tgccacctca gtgacatctc agggcaccga gactctggcc cttcagctgc
     3961 agaatgccga aacacagact gagagggagg tgccggagcc agggacagcc gcctcaggtc
     4021 ctggtgaagg tgagggttca gagtatggtg ccagtggaga agatgcgctc agcaggatcc
     4081 agaggctgat ggcggagggc ggcatgacag ccgtggtgca gcgggagcag agcaccacca
     4141 tggcctccat gggcggcttc ggcaacaaca tcatcgtcag ccaccgcatt caccgcagct
     4201 ctcagacggg cactgaacct ggtgccgccc acacctcctc accccagccc tccacctctc
     4261 ggggactgct cccagaggcc gggcaactgg cagagcgagg cctaagcccc cggacagctt
     4321 cctgggacca gcctggtacc cctgggcggg agccaaccca gccaaccctg ccctcttcct
     4381 cccctgtccc cattcctgtt tcccttccca gcgctgaggg accaaccgtc cactgcgagt
     4441 tgaccaataa caaccacctt ctggatggtg gcagcagcag gggggacgct gcaggcccta
     4501 ggggagaacc acggaacagg ggcggccgca tggactacaa ggaccacgac ggcgactaca
     4561 aggaccacga catcgactac aaggacgacg acgacaagta actcgagtct agagggcccg
     4621 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     4681 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     4741 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     4801 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     4861 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     4921 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4981 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5041 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5101 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     5161 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5221 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     5281 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5341 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     5401 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     5461 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     5521 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5581 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     5641 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     5701 ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat
     5761 atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc
     5821 gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc
     5881 gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc
     5941 ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg
     6001 gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg
     6061 gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc
     6121 ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg
     6181 ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     6241 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     6301 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     6361 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     6421 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     6481 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     6541 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     6601 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     6661 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     6721 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     6781 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6841 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6901 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6961 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     7021 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     7081 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     7141 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     7201 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     7261 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     7321 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7381 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     7441 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     7501 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     7561 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     7621 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     7681 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7741 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7801 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7861 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7921 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7981 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     8041 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     8101 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     8161 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     8221 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     8281 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     8341 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     8401 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     8461 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     8521 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     8581 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
