LOCUS       sequence-197034-        8636 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 8636)
  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 8636)
  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: pcdna3 flag - ULK1 wt
FEATURES             Location/Qualifiers
     source          1..8636
                     /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"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      660..669
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             669..692
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    705..729
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3887..3911)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(3955..3973)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3956..3973)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3993..4010)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3999..4223
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4269..4697
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4356..4375)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4566..4587
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4706..4726)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4711..5040
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4891..5026
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4953..4972
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5107..5901
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5161..5180)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5771..5790
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6075..6196
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6112..6131)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6166..6185
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6245..6261)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6245..6261)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6258..6280)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6269..6285
                     /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(6293..6299,6300..6317,6318..6323))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6338..6359
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6476..6493)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6647..7235)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6727..6746)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7406..8197,8198..8266))
                     /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     8029..8048
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8267..8371)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8446..8465)
                     /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 agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 ccaccatgga ctacaaggac gacgacgata aggaattcgt tatcacaagt ttgtacaaaa
      721 aagcaggctt cgagccgggc cgaggcggcg tcgagaccgt gggcaagttc gagttctctc
      781 gcaaggacct gattggacac ggcgccttcg cggtggtctt caagggtcga caccgcgaga
      841 agcacgacct ggaggtggcc gtcaaatgca ttaacaagaa gaaccttgcc aagtcccaaa
      901 cactgctggg aaaggaaatc aaaatcctga aggaactaaa gcacgaaaac atcgtggcgc
      961 tgtatgactt ccaggaaatg gctaattctg tctacctggt catggagtat tgtaatggtg
     1021 gagacctggc tgactacctg cacactatgc gcacactgag tgaagacact gtcaggcttt
     1081 tcctacagca gatcgctggc gccatgcggc tgctgcacag caagggcatc atccaccggg
     1141 acctgaagcc ccaaaacatc ctgctgtcca accctggggg ccgccgggcc aaccccagca
     1201 acatccgagt caagattgct gactttggat tcgctcggta cctccagagc aacatgatgg
     1261 cggccacact ctgtggttct cctatgtaca tggctcctga ggtcattatg tcccagcact
     1321 acgatggaaa ggctgacctg tggagcattg gcaccattgt ctaccagtgt ctgacaggga
     1381 aggccccttt tcaggccagc agccctcagg atttgcgcct gttttatgag aagaacaaga
     1441 cactagttcc tgccatcccc cgggagacat cagctcccct gcggcagctg ctcctggctc
     1501 tgttgcagcg gaaccacaag gaccgcatgg actttgatga atttttccac caccctttct
     1561 tggatgccag cacccccatc aagaaatccc cacctgtgcc tgtgccctca tatccaagct
     1621 cagggtctgg cagcagctcc agcagcagct ctgcctccca cctggcctct ccaccgtccc
     1681 tgggggagat gccacagcta cagaagaccc ttacctcccc agccgatgct gctggctttc
     1741 ttcagggctc ccgggactct ggtggcagca gcaaagactc ctgtgacaca gatgactttg
     1801 tcatggtccc agcccagttt ccaggtgatc tagttgctga ggcagccagt gccaagcccc
     1861 cacctgatag cctgctgtgt agtgggagct cattggtggc ctctgctggc ctagagagcc
     1921 acggccgtac cccctctccc tctccgacct gcagcagctc tcccagcccc tctggccggc
     1981 ctggcccctt ctccagcaac aggtacggtg cctcggtccc cattcctgtc cccactcagg
     2041 tgcacaatta ccagcgcatc gagcaaaacc tgcaatcgcc cactcaacag cagacagcca
     2101 ggtcctctgc catccgaagg tcagggagca ccagccccct gggctttggc cgggccagcc
     2161 catcaccccc ctcccacacc gatggagcca tgctggccag gaagctgtca cttggaggtg
     2221 gccgtcccta cacaccttct ccccaagtgg gaaccatccc agagcgaccc agctggagca
     2281 gagtgccctc cccacaagga gctgatgtgc gggttggcag gtcaccacga cccggttcct
     2341 ctgtgcctga gcactctcca agaaccactg ggctgggctg ccgcctgcac agtgccccta
     2401 acctgtccga cttccatgtt gtgcgtccca agctgcctaa gcccccaaca gacccactgg
     2461 gagccacctt tagcccaccc cagaccagcg caccccagcc atgcccaggg ctacagtctt
     2521 gccggccact gcgtggctca cctaagctgc ctgacttcct acagcggagt cccctacccc
     2581 ccatcctagg ctctcctacc aaggccgggc cctcctttga cttccccaaa acccccagct
     2641 ctcagaattt gctgaccctg ttggctaggc agggggtagt aatgacacca cctcggaacc
     2701 gtacactgcc tgacctctcc gaggccagtc ctttccatgg ccagcagctg ggctctggcc
     2761 ttcggcccgc tgaagacacc cggggtccct ttggacggtc cttcagcacc agccgcatta
     2821 cggacctgct gcttaaggct gcatttggga ctcaggcctc tgactcaggc agcacagaca
     2881 gcctacagga gaaacctatg gagattgctc cctctgctgg ctttggaggg actctgcatc
     2941 caggagctcg tggtggaggg gccagcagcc cagcacctgt ggtatttact gtaggctccc
     3001 cacccagtgg tgccacccca ccccagagta cccgtaccag aatgttctca gtgggctctt
     3061 ccagctccct gggctctact ggctcctcct ctgcccgcca cttagtgcct ggggcctgtg
     3121 gagaggcccc ggagctttct gccccaggcc actgctgtag ccttgctgac ccccttgctg
     3181 ccaacttgga gggggctgtg accttcgagg ctcctgacct cccagaggag accctcatgg
     3241 agcaagagca cacggaaacc ctacacagtc tgcgcttcac actagcgttt gcacagcaag
     3301 ttctggagat tgcagccctg aagggaagtg ccagtgaggc cgccggtggc cctgagtacc
     3361 agctccagga aagtgtggtg gctgaccaga tcagtcagtt gagccgagag tggggctttg
     3421 cagagcaact ggttctgtac ttgaaggtgg ctgagctgct gtcctcaggc ctacagactg
     3481 ccattgacca gattcgagct ggcaaactct gcctttcatc tactgtgaag caggtggtac
     3541 gcagactaaa tgagctgtac aaggccagcg tggtatcctg ccagggcctc agcttgcgac
     3601 ttcagcgctt ctttctggac aaacaacggc tgctggacgg gatccatggt gtcactgcag
     3661 agcggctcat cctcagccat gctgtgcaaa tggtacaatc agctgccctt gatgagatgt
     3721 tccagcaccg agagggctgt gtaccgagat atcacaaagc cctgctattg ctggaggggt
     3781 tgcagcacac tctcacggac caggcagaca ttgagaacat tgccaaatgc aagctgtgca
     3841 ttgagaggag actctcggcc ctgctgagtg gtgtctatgc ctgaataccc agctttcttg
     3901 tacaaagtgg tgataacgtg gatcccactc gagcatgcat ctagagggcc ctattctata
     3961 gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     4021 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     4081 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     4141 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     4201 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     4261 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4321 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4381 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4441 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4501 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4561 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4621 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4681 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4741 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4801 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4861 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4921 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4981 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5041 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     5101 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5161 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5221 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5281 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5341 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5401 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5461 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5521 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5581 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     5641 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5701 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5761 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5821 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5881 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5941 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     6001 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     6061 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     6121 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     6181 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     6241 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     6301 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     6361 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     6421 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     6481 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6541 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6601 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6661 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6721 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6781 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6841 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6901 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6961 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7021 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7081 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7141 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7201 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7261 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7321 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7381 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7441 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7501 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7561 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7621 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7681 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7741 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7801 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7861 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7921 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7981 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     8041 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     8101 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     8161 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     8221 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     8281 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     8341 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     8401 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     8461 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     8521 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     8581 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgtt
//
