LOCUS       sequence-191076-        7079 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of one subcomplex from the Human ULK1 complex.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7079)
  AUTHORS   Qi S, Kim DJ, Stjepanovic G, Hurley JH
  TITLE     Structure of the Human Atg13-Atg101 HORMA Heterodimer: an 
            Interaction Hub within the ULK1 Complex.
  JOURNAL   Structure. 2015 Oct 6;23(10):1848-57. doi: 
            10.1016/j.str.2015.07.011. Epub 2015 Aug 20.
  PUBMED    26299944
REFERENCE   2  (bases 1 to 7079)
  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: 
            "10.1016/j.str.2015.07.011"; journalName: "Structure"; date: 
            "2015-10-6- 6"; volume: "23"; issue: "10"; pages: "1848-57"
COMMENT     Sequence Label: pFastBac-ATG101-ATG13 HORMA
FEATURES             Location/Qualifiers
     source          1..7079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  1..166
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    195..329
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(219..238)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     273..292
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(579..590)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             complement(1096..1116)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(1129..1782)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(1149..1171)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     promoter        complement(1840..1931)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(1898..1915)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     promoter        1955..2064
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     primer_bind     complement(2746..2765)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2790..2838
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        join(3346..3351,3352..3368,3369..3374)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3563..4096
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  4163..4387
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(4521..4538)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4692..5280)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4772..4791)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5451..6242,6243..6311))
                     /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     6074..6093
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6312..6416)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6442..6896)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6579..6600)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6790..6809
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tgtgggcgga caaaatagtt gggaactggg aggggtggaa atggagtttt taaggattat
       61 ttagggaaga gtgacaaaat agatgggaac tgggtgtagc gtcgtaagct aatacgaaaa
      121 ttaaaaatga caaaatagtt tggaactaga tttcacttat ctggttcgga tctcctaggc
      181 tcaagcagtg atcagatcca gacatgataa gatacattga tgagtttgga caaaccacaa
      241 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
      301 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
      361 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
      421 tggctgatta tgatcctcta gtacttctcg acaagcttgt cgagactgca ggctctagat
      481 tcgaaagcgg ccgcgactag tgagctcgtc gacttactgt ctggttgaca tgaaggccag
      541 attgatacgg tatgcgcaag agagggtaat tgtgccgact ggagttccca cggtaccgac
      601 acgcacagtc tgaaagcctt cacccagtcc ggagagctgg acctcgccga aatagatcct
      661 gtacagaatc acatattcat gaccctgctt cctactgagg cggtaagcag gtgtcacccg
      721 agtgatggcc aggagagatt tcaggagcag ggacagacga ttatagacgg tgtagctcac
      781 tttgatctct ttatcacact tttcgttcat ctccaggcac caaatttcga gctccattga
      841 gtctccttca gatgtcttca gggagatctc gacacacatt gaacgaccca ctgctggcag
      901 ctgtccagcg agtgctttct ttgcttcgtg agtcacttca gggatatcct taatagccag
      961 gttgaaccag tcagaaccgg tgggtgaaga ggagctgcgt gtgcagattt tctctcccag
     1021 ccgggcctgc acgatcacct gcacggtctt gagggcgaaa aacttaatga acttgtcgag
     1081 gtcggatccc atggcgccct gaaaatacag gttttcggtc gtatcgattt ttggaggatg
     1141 gtcgccacca ccaaacgtgg cttgccagcc ctgcaaaggc catgctatat acttgctgga
     1201 tttcaagtac ttatcaattt gtgggatagc ttcaatacgt tttttaaaac aaactaattt
     1261 tgggaacgca tccaggcaca ttgggtccat gtataaaaca acatcaagag cgtcatacaa
     1321 catgaagtca ggatgggtta catgatcacc atttaaatat gttttatgac ataaacgatc
     1381 ttcgaacatt ttcagcattt caggtagctt gctaagaaaa tcaactttga gagtttcaaa
     1441 gtctttacta tatgcaattc tcgaaacacc gtatctaata tccaaaaccg ctccttcaag
     1501 cattgaaatc tctgcacgct cttttggaca accacccaac atgttgtgct tgtcagctat
     1561 ataacgtatg atggccatag actgtgttaa tttaacatca ccatcaatat aataaggaag
     1621 attgggaaac tccaaaccca attcaaactt tttgtttcgc catttatcac cttcatcgcg
     1681 ctcatacaaa tgctcttcat atttttcttc aagatattcc aaaagaagtc gagtgggttg
     1741 cacaaggccc ttaattttcc aataacctag tataggggac atgcgcgctt cggaccggga
     1801 tccgcgcccg atggtgggac ggtatgaata atccggaata tttataggtt tttttattac
     1861 aaaactgtta cgaaaacagt aaaatactta tttatttgcg agatggttat cattttaatt
     1921 atctccatga tctattaata ttccggagta tacggacctt taattcaacc caacacaata
     1981 tattatagtt aaataagaat tattatcaaa tcatttgtat attaattaaa atactatact
     2041 gtaaattaca ttttatttac aatcactcga cgaagacttg atcacccggg atctcgacat
     2101 gaattgccgg tcagaggtcc tggaagtgag cgtggaaggt agacaggtgg aagaggctat
     2161 gctggccgtg ctgcatacag tgctgctgca cagatcaact ggcaagttcc attacaagaa
     2221 agaaggaaca tattccatcg gaaccgtggg tactcaggac gtcgattgcg acttcattga
     2281 ttttacttac gtgcgcgtca gctccgagga actggacaga gcactccgta aggtggtcgg
     2341 agaattcaaa gatgccctga ggaacagtgg cggagacggt ctgggccaga tgagcctcga
     2401 gttctatcag aagaaaaagt ctagatggcc attttcagat gaatgtatcc cctgggaagt
     2461 gtggaccgtg aaggtccacg tggtcgccct ggctactgag caggaacggc agatctgccg
     2521 cgagaaagtg ggcgaaaagc tctgtgagaa gatcatcaac atcgtggaag tcatgaaccg
     2581 tcatgagtac ctgcccaaga tgcctacaca gagcgaggtg gataatgtct tcgacaccgg
     2641 tctgcgggat gtgcagccct acctctataa gatctccttt cagattacag actaagctag
     2701 cagctgatgc atagcatgcg gtaccgggag atgggggagg ctaactgaaa cacggaagga
     2761 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     2821 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     2881 ccccaccgag accccattgg gaccaatacg cccgcgtttc ttccttttcc ccaccccaac
     2941 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca agccctgcca
     3001 tagccactac gggtacgtag ccaaccacta gaactatagc tagagtcctg ggcgaacaaa
     3061 cgatgctcgc cttccagaaa accgaggatg cgaaccactt catccggggt cagcaccacc
     3121 ggcaagcgcc gcgacggccg aggtctaccg atctcctgaa gcaagggcag atccgtgcac
     3181 agcaccttgc cgtagaagaa cagcaaggcc gccaatgcct gacgatgcgt ggagaccgaa
     3241 accttgcgct cgttcgccag ccaggacaga aatgcctcga cttcgctgct gcccaaggtt
     3301 gccgggtgac gcacaccgtg gaaacggatg aaggcacgaa cccagttgac ataagcctgt
     3361 tcggttcgta aactgtaatg caagtagcgt atgcgctcac gcaactggtc cagaaccttg
     3421 accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt
     3481 tttgtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt
     3541 ttgatgttat ggagcagcaa cgatgttacg cagcagcaac gatgttacgc agcagggcag
     3601 tcgccctaaa acaaagttag gtggctcaag tatgggcatc attcgcacat gtaggctcgg
     3661 ccctgaccaa gtcaaatcca tgcgggctgc tcttgatctt ttcggtcgtg agttcggaga
     3721 cgtagccacc tactcccaac atcagccgga ctccgattac ctcgggaact tgctccgtag
     3781 taagacattc atcgcgcttg ctgccttcga ccaagaagcg gttgttggcg ctctcgcggc
     3841 ttacgttctg cccaagtttg agcagccgcg tagtgagatc tatatctatg atctcgcagt
     3901 ctccggcgag caccggaggc agggcattgc caccgcgctc atcaatctcc tcaagcatga
     3961 ggccaacgcg cttggtgctt atgtgatcta cgtgcaagca gattacggtg acgatcccgc
     4021 agtggctctc tatacaaagt tgggcatacg ggaagaagtg atgcactttg atatcgaccc
     4081 aagtaccgcc acctaacaat tcgttcaagc cgagatcggc ttcccggccg cggagttgtt
     4141 cggtaaattg tcacaacgcc gcgaatatag tctttaccat gcccttggcc acgcccctct
     4201 ttaatacgac gggcaatttg cacttcagaa aatgaagagt ttgctttagc cataacaaaa
     4261 gtccagtatg ctttttcaca gcataactgg actgatttca gtttacaact attctgtcta
     4321 gtttaagact ttattgtcat agtttagatc tattttgttc agtttaagac tttattgtcc
     4381 gcccacaccc gcttacgcag ggcatccatt tattactcaa ccgtaaccga ttttgccagg
     4441 ttacgcggct ggtctatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     4501 atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4561 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4621 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4681 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4741 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4801 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4861 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4921 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4981 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5041 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5101 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5161 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5221 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5281 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5341 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5401 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5461 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5521 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5581 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5641 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5701 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5761 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5821 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5881 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5941 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6001 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6061 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6121 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6181 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6241 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6301 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6361 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6421 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     6481 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     6541 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     6601 tattaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     6661 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     6721 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     6781 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     6841 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca
     6901 ttcgccattc aggctgcaaa taagcgttga tattcagtca attacaaaca ttaataacga
     6961 agagatgaca gaaaaatttt cattctgtga cagagaaaaa gtagccgaag atgacggttt
     7021 gtcacatgga gttggcagga tgtttgatta aaaacataac aggaagaaaa atgccccgc
//
