LOCUS       40924_16575        7719 bp DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7719)
  AUTHORS   Egan DF, Shackelford DB, Mihaylova MM, Gelino SR, 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. 2010 Dec 23. ():.
  PUBMED    21205641
REFERENCE   2  (bases 1 to 7719)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7719)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science. 
            2010 Dec 23. ():."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /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)."
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        253..463
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        558..1736
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             1788..2441
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="MAPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             2448..2465
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     primer_bind     complement(4848..4864)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5077..5532
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5549..5568)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     5668..5690
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5728..5746)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        5814..5918
                     /label=AmpR promoter
     CDS             5919..6776
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6950..7538
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7692..7709
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctta
      241 attccctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
      301 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
      361 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
      421 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctttgc
      481 aaagatggat aaagttttaa acagagagga atctttgcag ctaatggacc ttctaggtct
      541 tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
      601 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
      661 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
      721 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
      781 gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc
      841 cttgcgtgcc ttgaattact tccacctggc tgcagtacgt gattcttgat cccgagcttc
      901 gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg
      961 cttgagttga ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc
     1021 gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg
     1081 cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta
     1141 tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg
     1201 cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc
     1261 ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc
     1321 tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag
     1381 ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa
     1441 ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc
     1501 cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg
     1561 aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag
     1621 cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca
     1681 ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgaggaa
     1741 ttctctagag atccctcgac ctcgagatcc attgtgctgg atctacaatg gcccctatac
     1801 taggttattg gaaaattaag ggccttgtgc aacccactcg acttcttttg gaatatcttg
     1861 aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa
     1921 agtttgaatt gggtttggag tttcccaatc ttccttatta tattgatggt gatgttaaat
     1981 taacacagtc tatggccatc atacgttata tagctgacaa gcacaacatg ttgggtggtt
     2041 gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc ggttttggat attagatacg
     2101 gtgtttcgag aattgcatat agtaaagact ttgaaactct caaagttgat tttcttagca
     2161 agctacctga aatgctgaaa atgttcgaag atcgtttatg tcataaaaca tatttaaatg
     2221 gtgatcatgt aacccatcct gacttcatgt tgtatgacgc tcttgatgtt gttttataca
     2281 tggacccaat gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag
     2341 ctatcccaca aattgataag tacttgaaat ccagcaagta tatagcatgg cctttgcagg
     2401 gctggcaagc cacgtttggt ggtggcgacc atcctccaaa atcggatctg gttccgcgtg
     2461 gatccactag tggtaccatc gatgcggccg ctaattcggc acgagccgag aagcagaagc
     2521 acgacgggcg ggtgaagatc ggccactaca tcctggggga cacgctgggc gtcggcacct
     2581 tcgggaaagt gaaggtgggc aagcacgagt tgactggaca taaagttgct gtgaagatac
     2641 tcaaccggca gaagattcga agcctggacg tggtcgggaa aatccgcaga gagatccaga
     2701 acctgaagct tttcaggcac cctcatataa tcaaactgta ccaggtcatc agtacaccgt
     2761 ctgatatttt catggtcatg gaatatgtct caggaggaga gctatttgat tatatctgta
     2821 aaaatggaag gttggacgaa aaggagagtc gacgtctgtt ccagcagatc ctttctggtg
     2881 tggactattg tcacaggcat atggtggtcc acagagattt gaaacctgaa aacgtcctgc
     2941 ttgatgcaca catgaatgca aagatagccg acttcggtct ttcaaacatg atgtcagatg
     3001 gtgaattttt aagaacgagc tgtggctcgc ccaattatgc tgcaccagaa gtaatttcag
     3061 gaagattgta cgcaggccct gaagtagaca tctggagcag cggggtcatt ctctatgctt
     3121 tgctgtgtgg aactctccct tttgatgatg accacgtgcc aactcttttt aagaagatat
     3181 gtgacgggat attttatacc cctcagtatt tgaatccctc tgtaataagc cttttgaagc
     3241 atatgctgca ggtagatcct atgaagaggg ccacaataaa agatatcagg gaacatgaat
     3301 ggtttaagca ggaccttcca aaatatctct ttcctgaaga cccgtcttat agttcaacca
     3361 tgattgatga tgaagcctta aaagaagtgt gtgagaagtt cgagtgctca gaggaggagg
     3421 tcctcagctg cctgtactaa ggcgcgcctt agtacaacag aaaccaccag gacccactgg
     3481 cagttgccta ccacctcata atagacaaca ggagaataat gaacgaagcc aaagatttct
     3541 acttggcaac aagcccaccc gattctttcc tcgatgatca ccatttaact cggcctcacc
     3601 ctgagagagt accattcttg gttgccgaaa caccaagggc ccgacacacc ctagatgaat
     3661 taaacccaca gaaatccaaa caccaaggcg tacggaaggc aaagtggcat ttggggattc
     3721 gaagtcaaag ccgacccaat gacatcatgg cagaagtgtg tagagcaatc aagcagttgg
     3781 actatgaatg gaaggttgta aacccctatt atttgcgtgt gcgaaggaag aaccctgtga
     3841 caagcacatt ttccaaaatg agtctacagc tataccaagt ggatagtagg acttacttat
     3901 tggatttccg aagtattgat gatgagatta cagaagccaa atcagggact gctactccac
     3961 agagatcggg atccatcagc aactatcgat cttgccaaag gagcgactcc gacgccgagg
     4021 ctcaaggaaa gccctcagaa gtctctctta cctcatccgt gacctccctc gactcctctc
     4081 ctgttgacgt agctccaaga ccaggaagtc acacgataga attttttgaa atgtgtgcaa
     4141 atctaattaa aattcttgca cagtaaacct aaaactttgc ttatttcttt gatgcaataa
     4201 gagcggccgc gactctagag tgagggtccc cacctgggac ccttgagagt atcaggtctc
     4261 ccacgtggga gacaagaaat ccctgtttaa tatttaaaca gcagtgttcc ccatctgggt
     4321 ccttgcaccc ctcactctgg cctcagccga ctgcacagcg gcccctgcat ccccttggct
     4381 gtgaggcccc tggacaagca gaggtggcca gagctgggag gcatggccct ggggtcccac
     4441 gaatttgctg gggaatctcg tttttcttct taagactttt gggacatggt ttgactcccg
     4501 aacatcaccg acgtgtctcc tgtttttctg ggtggcctcg ggacacctgc cctgccccca
     4561 cgagggtcag gactgtgact ctttttaggg ccaggcaggt gcctggacat ttgccttgct
     4621 ggacggggac tggggatgtg ggagggagca gacaggagga atcatgtcag gcctgtgtgt
     4681 gaaaggaagc tccactgtca ccctccacct cttcaccccc cactcaccag tgtcccctcc
     4741 actgtcacat tgtaactgaa cttcaggata ataaagtgtt tgcctccaaa aaaaaaaaaa
     4801 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaac cgaattcact ggccgtcgtt
     4861 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     4921 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     4981 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
     5041 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     5101 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     5161 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5221 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5281 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5341 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5401 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     5461 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     5521 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5581 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     5641 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     5701 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5761 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5821 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5881 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5941 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     6001 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     6061 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     6121 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     6181 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     6241 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6301 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     6361 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6421 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     6481 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     6541 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6601 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6661 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6721 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6781 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6841 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6901 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6961 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     7021 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     7081 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     7141 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     7201 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     7261 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7321 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7381 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7441 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     7501 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     7561 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     7621 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     7681 cgaccgagcg cagcgagtca gtgagcgagg aagcggaag
//
