LOCUS       sequence-419744-        8359 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 8359)
  AUTHORS   Egan DF, Shackelford DB, Mihaylova MM, Gelino S, Kohnz RA, Mair W, 
            Vasquez DS, Joshi A, Gwinn DM, Taylor R, Asara JM, Fitzpatrick J, 
            Dillin A, Viollet B, Kundu M, Hansen M, Shaw RJ
  TITLE     Phosphorylation of ULK1 (hATG1) by AMP-activated protein kinase 
            connects energy sensing to mitophagy.
  JOURNAL   Science. 2011 Jan 28;331(6016):456-61. doi: 10.1126/science.1196371.
            Epub 2010 Dec 23.
  PUBMED    21205641
REFERENCE   2  (bases 1 to 8359)
  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: 
            "10.1126/science.1196371"; journalName: "Science"; date: 
            "2011-01-28- 28"; volume: "331"; issue: "6016"; pages: "456-61"
COMMENT     Sequence Label: pcdna6.2- myc ULK1 4SA
FEATURES             Location/Qualifiers
     source          1..8359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        42..421
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        422..625
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     576..596
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     670..689
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        670..688
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    722..746
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             755..784
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(3949..3973)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3987..4028
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4047..4064
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     primer_bind     complement(4104..4123)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4148..4196
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4398..4826
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4485..4504)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4695..4716
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4835..4855)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4840..5169
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5020..5155
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5082..5101
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5217..5264
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5283..5681
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5839..5960
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5876..5895)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5930..5949
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6009..6025)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6009..6025)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6022..6044)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6033..6049
                     /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(6057..6063,6064..6081,6082..6087))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6102..6123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6240..6257)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6411..6999)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6491..6510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7170..7961,7962..8030))
                     /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     7793..7812
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8031..8135)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8210..8229)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
       61 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      121 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      181 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      241 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      301 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      361 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      421 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      481 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      541 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      601 gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg cttactggct
      661 tatcgaaatt aatacgactc actataggga gacccaagct ggctagttaa gctgagcatc
      721 aacaagtttg tacaaaaaag caggcttcac catggaacaa aagctaatct ccgaggaaga
      781 cttgggagga atagagccgg gccgaggcgg cgtcgagacc gtgggcaagt tcgagttctc
      841 tcgcaaggac ctgattggac acggcgcctt cgcggtggtc ttcaagggtc gacaccgcga
      901 gaagcacgac ctggaggtgg ccgtcaaatg cattaacaag aagaaccttg ccaagtccca
      961 aacactgctg ggaaaggaaa tcaaaatcct gaaggaacta aagcacgaaa acatcgtggc
     1021 gctgtatgac ttccaggaaa tggctaattc tgtctacctg gtcatggagt attgtaatgg
     1081 tggagacctg gctgactacc tgcacactat gcgcacactg agtgaagaca ctgtcaggct
     1141 tttcctacag cagatcgctg gcgccatgcg gctgctgcac agcaagggca tcatccaccg
     1201 ggacctgaag ccccaaaaca tcctgctgtc caaccctggg ggccgccggg ccaaccccag
     1261 caacatccga gtcaagattg ctgactttgg attcgctcgg tacctccaga gcaacatgat
     1321 ggcggccaca ctctgtggtt ctcctatgta catggctcct gaggtcatta tgtcccagca
     1381 ctacgatgga aaggctgacc tgtggagcat tggcaccatt gtctaccagt gtctgacagg
     1441 gaaggcccct tttcaggcca gcagccctca ggatttgcgc ctgttttatg agaagaacaa
     1501 gacactagtt cctgccatcc cccgggagac atcagctccc ctgcggcagc tgctcctggc
     1561 tctgttgcag cggaaccaca aggaccgcat ggactttgat gaatttttcc accacccttt
     1621 cttggatgcc agcaccccca tcaagaaatc cccacctgtg cctgtgccct catatccaag
     1681 ctcagggtct ggcagcagct ccagcagcag ctctgcctcc cacctggcct ctccaccgtc
     1741 cctgggggag atgccacagc tacagaagac ccttacctcc ccagccgatg ctgctggctt
     1801 tcttcagggc tcccgggact ctggtggcag cagcaaagac tcctgtgaca cagatgactt
     1861 tgtcatggtc ccagcccagt ttccaggtga tctagttgct gaggcagcca gtgccaagcc
     1921 cccacctgat agcctgctgt gtagtgggag ctcattggtg gcctctgctg gcctagagag
     1981 ccacggccgt accccctctc cctctccgac ctgcagcagc tctcccagcc cctctggccg
     2041 gcctggcccc ttctccagca acaggtacgg tgcctcggtc cccattcctg tccccactca
     2101 ggtgcacaat taccagcgca tcgagcaaaa cctgcaatcg cccactcaac agcagacagc
     2161 caggtcctct gccatccgaa ggtcaggggc caccagcccc ctgggctttg gccgggccag
     2221 cccatcaccc ccctcccaca ccgatggagc catgctggcc aggaagctgt cacttggagg
     2281 tggccgtccc tacacacctt ctccccaagt gggaaccatc ccagagcgac ccagctggag
     2341 cagagtgccc tccccacaag gagctgatgt gcgggttggc aggtcaccac gacccggttc
     2401 ctctgtgcct gagcactctc caagaaccac tgggctgggc tgccgcctgc acgctgcccc
     2461 taacctgtcc gacttccatg ttgtgcgtcc caagctgcct aagcccccag cagacccact
     2521 gggagccacc tttagcccac cccagaccag cgcaccccag ccatgcccag ggctacagtc
     2581 ttgccggcca ctgcgtggct cacctaagct gcctgacttc ctacagcgga gtcccctacc
     2641 ccccatccta ggctctccta ccaaggccgg gccctccttt gacttcccca aaacccccgc
     2701 ctctcagaat ttgctgaccc tgttggctag gcagggggta gtaatgacac cacctcggaa
     2761 ccgtacactg cctgacctct ccgaggccag tcctttccat ggccagcagc tgggctctgg
     2821 ccttcggccc gctgaagaca cccggggtcc ctttggacgg tccttcagca ccagccgcat
     2881 tacggacctg ctgcttaagg ctgcatttgg gactcaggcc tctgactcag gcagcacaga
     2941 cagcctacag gagaaaccta tggagattgc tccctctgct ggctttggag ggactctgca
     3001 tccaggagct cgtggtggag gggccagcag cccagcacct gtggtattta ctgtaggctc
     3061 cccacccagt ggtgccaccc caccccagag tacccgtacc agaatgttct cagtgggctc
     3121 ttccagctcc ctgggctcta ctggctcctc ctctgcccgc cacttagtgc ctggggcctg
     3181 tggagaggcc ccggagcttt ctgccccagg ccactgctgt agccttgctg acccccttgc
     3241 tgccaacttg gagggggctg tgaccttcga ggctcctgac ctcccagagg agaccctcat
     3301 ggagcaagag cacacggaaa ccctacacag tctgcgcttc acactagcgt ttgcacagca
     3361 agttctggag attgcagccc tgaagggaag tgccagtgag gccgccggtg gccctgagta
     3421 ccagctccag gaaagtgtgg tggctgacca gatcagtcag ttgagccgag agtggggctt
     3481 tgcagagcaa ctggttctgt acttgaaggt ggctgagctg ctgtcctcag gcctacagac
     3541 tgccattgac cagattcgag ctggcaaact ctgcctttca tctactgtga agcaggtggt
     3601 acgcagacta aatgagctgt acaaggccag cgtggtatcc tgccagggcc tcagcttgcg
     3661 acttcagcgc ttctttctgg acaaacaacg gctgctggac gggatccatg gtgtcactgc
     3721 agagcggctc atcctcagcc atgctgtgca aatggtacaa tcagctgccc ttgatgagat
     3781 gttccagcac cgagagggct gtgtaccgag atatcacaaa gccctgctat tgctggaggg
     3841 gttgcagcac actctcacgg accaggcaga cattgagaac attgccaaat gcaagctgtg
     3901 cattgagagg agactctcgg ccctgctgag tggtgtctat gcctgaatac ccagctttct
     3961 tgtacaaagt ggttgatgct gttaacggga agcctatccc taaccctctc ctcggtctcg
     4021 attctacgcg taccggtgct ggtggctgtt gtcctggctg ttgcggtggc ggctagtaat
     4081 gagtttaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     4141 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     4201 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     4261 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     4321 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     4381 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4441 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4501 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4561 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4621 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4681 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4741 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4801 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4861 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4921 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4981 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5041 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5101 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     5161 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg
     5221 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
     5281 ccatggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa
     5341 tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg
     5401 gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg
     5461 tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg
     5521 gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc
     5581 tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga
     5641 ttcgtgaatt gctgccctct ggttatgtgt gggagggcta agcacttcgt ggccgaggag
     5701 caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5761 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5821 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5881 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5941 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     6001 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6061 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6121 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6181 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6241 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6301 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6361 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6421 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6481 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6541 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6601 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6661 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6721 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6781 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6841 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6901 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6961 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7021 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7081 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7141 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7201 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7261 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7321 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7381 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7441 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7501 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7561 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7621 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7681 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7741 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7801 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7861 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7921 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7981 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8041 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8101 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8161 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     8221 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     8281 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     8341 atctgcttag ggttaggcg
//
