LOCUS       sequence-370105-        5583 bp    DNA     circular SYN 15-APR-2025
DEFINITION  To make ATG101/ATG13 HORMA complex.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5583)
  AUTHORS   Shi X, Yokom AL, Wang C, Young LN, Youle RJ, Hurley JH
  TITLE     ULK complex organization in autophagy by a C-shaped FIP200 
            N-terminal domain dimer.
  JOURNAL   J Cell Biol. 2020 Jul 6;219(7):e201911047. doi: 
            10.1083/jcb.201911047.
  PUBMED    32516362
REFERENCE   2  (bases 1 to 5583)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Biol."; date: "2020-07-6"; volume: "219(7):e201911047. doi"; pages: 
            " 10.1083/jcb.201911047"
COMMENT     Sequence Label: pCAG-ATG101
FEATURES             Location/Qualifiers
     source          1..5583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          738..1754
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1678..1700
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1762..1781
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(2605..2624)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2670..2725
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2724..2743)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3086..3102)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3086..3102)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3099..3121)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3110..3126
                     /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(3134..3140,3141..3158,3159..3164))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3179..3200
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3259..3454
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3305..3440
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3460..3594
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3497..3516)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3551..3570
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3662..3679)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3833..4421)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3913..3932)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4592..5383,5384..5452))
                     /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     5215..5234
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5453..5557)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gccccrgggg gggggggggg gggggggggg
      541 gggggggggg gggggggggg gggggggggg gggggggggg gcgcgcgcsg ggsggggygg
      601 ggyggggcgr ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg
      661 gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc
      721 gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg
      781 ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc
      841 gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt
      901 ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg
      961 gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc
     1021 ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg
     1081 aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag
     1141 gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc
     1201 tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg
     1261 ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg
     1321 gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg
     1381 gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt
     1441 aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg
     1501 ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga
     1561 aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc
     1621 agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt
     1681 tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct
     1741 tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca
     1801 aagaattcct cgatcgacgg tatcgatgcc accatgaatt gccggtcaga ggtcctggaa
     1861 gtgagcgtgg aaggtagaca ggtggaagag gctatgctgg ccgtgctgca tacagtgctg
     1921 ctgcacagat caactggcaa gttccattac aagaaagaag gaacatattc catcggaacc
     1981 gtgggtactc aggacgtcga ttgcgacttc attgatttta cttacgtgcg cgtcagctcc
     2041 gaggaactgg acagagcact ccgtaaggtg gtcggagaat tcaaagatgc cctgaggaac
     2101 agtggcggag acggtctggg ccagatgagc ctcgagttct atcagaagaa aaagtctaga
     2161 tggccatttt cagatgaatg tatcccctgg gaagtgtgga ccgtgaaggt ccacgtggtc
     2221 gccctggcta ctgagcagga acggcagatc tgccgcgaga aagtgggcga aaagctctgt
     2281 gagaagatca tcaacatcgt ggaagtcatg aaccgtcatg agtacctgcc caagatgcct
     2341 acacagagcg aggtggataa tgtcttcgac accggtctgc gggatgtgca gccctacctc
     2401 tataagatct cctttcagat tacagacgct ctgggcacct ctgtcaccac tacaatgcgc
     2461 aggctcatca aagacaccct ggcactctaa cccgggtgat atcttaatta attaactcga
     2521 ggaattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc
     2581 cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat
     2641 gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt
     2701 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa
     2761 acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat
     2821 gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc
     2881 cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt
     2941 atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct
     3001 cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct
     3061 gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3121 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3181 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3241 tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3301 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3361 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3421 aggaggcttt tttggaggcc taggcttttg caaaaagcta acttgtttat tgcagcttat
     3481 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3541 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgctgc
     3601 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3661 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3721 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3781 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3841 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3901 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3961 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4021 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4081 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4141 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4201 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4261 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4321 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4381 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4441 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4501 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4561 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4621 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4681 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4741 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4801 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4861 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4921 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4981 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5041 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5101 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5161 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5221 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5281 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5341 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5401 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5461 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5521 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5581 ctg
//
