LOCUS       sequence-200647-        7528 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of  dATG1 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7528)
  AUTHORS   Kim M, Park HL, Park HW, Ro SH, Nam SG, Reed JM, Guan JL, Lee JH
  TITLE     Drosophila Fip200 is an essential regulator of autophagy that 
            attenuates both growth and aging.
  JOURNAL   Autophagy. 2013 Aug;9(8):1201-13. doi: 10.4161/auto.24811. Epub 2013
            May 14.
  PUBMED    23819996
REFERENCE   2  (bases 1 to 7528)
  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: 
            "10.4161/auto.24811"; journalName: "Autophagy"; date: "2013-08"; 
            volume: "9"; issue: "8"; pages: "1201-13"
COMMENT     Sequence Label: pGEX-4T-1 dATG1 (1188)
FEATURES             Location/Qualifiers
     source          1..7528
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     46..63
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        136..213
                     /gene="lacI"
                     /label=lacI promoter
     CDS             214..1296
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(233..252)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1345..1350,1351..1368,1369..1375)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1383..1399
                     /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)."
     primer_bind     1407..1423
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1407..1423
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1419..1946
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1439..1456)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1439..1455)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1448..1470)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2048..2053,2054..2069,2070..2076)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2084..2100
                     /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)."
     CDS             2123..2776
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2734..2756
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2783..2800
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(3952..3963)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(5443..5465)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5610..5628)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5696..5800
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5801..5869,5870..6661)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6019..6038)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6832..7420
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7321..7340
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
       61 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
      121 caccgcataa attccgacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag
      181 cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc
      241 gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac
      301 gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc
      361 aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc
      421 agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa
      481 ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg
      541 gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat
      601 gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat
      661 gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg
      721 ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta
      781 agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa
      841 attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc
      901 atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg
      961 gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg
     1021 gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc
     1081 aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag
     1141 ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc
     1201 ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     1261 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta
     1321 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     1381 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acggattcac
     1441 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     1501 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     1561 cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt ccggcaccag
     1621 aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga ggccgatact gtcgtcgtcc
     1681 cctcaaactg gcagatgcac ggttacgatg cgcccatcta caccaacgtg acctatccca
     1741 ttacggtcaa tccgccgttt gttcccacgg agaatccgac gggttgttac tcgctcacat
     1801 ttaatgttga tgaaagctgg ctacaggaag gccagacgcg aattattttt gatggcgttg
     1861 gaattagctt atcgactgca cggtgcacca atgcttctgg cgtcaggcag ccatcggaag
     1921 ctgtggtatg gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc aaggcgcact
     1981 cccgttctgg ataatgtttt ttgcgccgac atcataacgg ttctggcaaa tattctgaaa
     2041 tgagctgttg acaattaatc atcggctcgt ataatgtgtg gaattgtgag cggataacaa
     2101 tttcacacag gaaacagtat tcatgtcccc tatactaggt tattggaaaa ttaagggcct
     2161 tgtgcaaccc actcgacttc ttttggaata tcttgaagaa aaatatgaag agcatttgta
     2221 tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt gaattgggtt tggagtttcc
     2281 caatcttcct tattatattg atggtgatgt taaattaaca cagtctatgg ccatcatacg
     2341 ttatatagct gacaagcaca acatgttggg tggttgtcca aaagagcgtg cagagatttc
     2401 aatgcttgaa ggagcggttt tggatattag atacggtgtt tcgagaattg catatagtaa
     2461 agactttgaa actctcaaag ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt
     2521 cgaagatcgt ttatgtcata aaacatattt aaatggtgat catgtaaccc atcctgactt
     2581 catgttgtat gacgctcttg atgttgtttt atacatggac ccaatgtgcc tggatgcgtt
     2641 cccaaaatta gtttgtttta aaaaacgtat tgaagctatc ccacaaattg ataagtactt
     2701 gaaatccagc aagtatatag catggccttt gcagggctgg caagccacgt ttggtggtgg
     2761 cgaccatcct ccaaaatcgg atctggttcc gcgtggatcc ccggaattca tgaatattgt
     2821 cggggaatat gaatacagct ccaaggatat gctgggccat ggcgcctttg ccgtcgtcta
     2881 caaaggacgt catcgcaaga aacacatgcc ggtggccatc aagtgcatca cgaagaaggg
     2941 acaactgaag acgcagaatc tgctcggcaa ggagatcaag atcctgaagg agctcaccga
     3001 gctgcaccat gagaatgtgg tggctctgct ggactgcaag gagtcccagg attgcgtcag
     3061 cctggtcatg gagtattgca atggcggcga cttggcggat tatctgagtg tcaaggggac
     3121 gctcagcgag gataccgtta gactcttcct cgtgcaacta gctggtgcta tgaaagcact
     3181 ttataccaaa ggaattgtgc atcgtgatct caagccacaa aacattttgc tatcgcacaa
     3241 ttatggcaaa acattgccag ctccatcgaa aataaccctg aaaattgcgg attttgggtt
     3301 tgcgcgattc ctgaacgagg gcgccatggc agccactctg tgcggctctc ccatgtatat
     3361 ggcgcccgag gtgatcatgt cgctgcagta cgactccaag gcggatctct ggtcgctggg
     3421 aacgattgtc taccagtgtt tgactggcaa ggcgccattc tatgcacaaa cgcccaacga
     3481 actgaagtcc tactacgagc agaatgctaa cctggcacca aaaattccaa gtggcgtgtc
     3541 accggatctg agggatctgc tgctgtgcct gctgcgccgc aactccaagg atcgcatctc
     3601 gtacgagagc ttctttgttc accgcttcct tcagggcaag aaggcggccg tgtcgccagg
     3661 tgagcctagt caagcagcca acctgcgacg acgagtctct aagagtggtg ttcttccagt
     3721 tgacatgcca ccgcttggcg gaacaccacc tgccaaggcc aagagcccct tgcagcagca
     3781 gctggagcag gaactgaagc tggtcaagct ggccgagcaa cagcagaagg agcgcgagga
     3841 gcaggaggcc caggaggacg agaacacggt gtccgtggtg gccaatccgg ccatttgtgc
     3901 caccatcacc aacgtaggtg tcctgtgcga cagtgagaac aacagcggtt cttgcagctc
     3961 ccacgaggac tccgatgact ttgtgctggt gccaaagaac cttccggagg accagcgaca
     4021 gggtctggcc caagtccaag cccagcccgc ttccggtggc caacgtccac agcagcaaca
     4081 aaatcaatcc agtccgccac gtcccagcag tctgcccatc tctgaaccca aaccagtacc
     4141 ggctcccgct cggcggcagg tggccaggcc cgggccgtta acggtagcca cactgggtgg
     4201 ccagcagata ccccgatcgc agccaattag cgtaaagcaa ccgcgaccgg atcagcgcaa
     4261 gagcagtgtg agcagtgaca ttaactcgat ctcaccgccg gcggtgcagt ttgccattgg
     4321 aacgccgcct acacgtatgc gttctgcctc tggtggatcg ctgtcggaga caccgccgcc
     4381 gcatgcaccg agcacctggc aggtgtcgcc aggacactcg caatcgccgt tgcgtcgaag
     4441 cggcaatagc tcgccagttc tgccatccgc tgcgcttacc aaattgccca ctttgggcag
     4501 tccaacaatg ctggtggcgc cgggatcatt gggctcgatt ggttcggcgg gcagtggatc
     4561 ggagaacaac aatcagcatc acatgctggg accgcgagct tttacgctgc ccgaactggg
     4621 ggccactggt ggcctacata gtctgctgga cacaggagcc ggcggcggtg gggaaccaca
     4681 tgcattccag gccccagaac tgtccgaaga gacgctgatg gatcgcgagc acaacgagac
     4741 attgtccaag ctgaactttg tgctggcgct caccgattgt atccaggaag ttgccgattc
     4801 gcgatgcgca ccgctctcca cgttcatggt ggccggcagt cagtcggcgg cgcaagcggc
     4861 gtcagcggat gcacagcaga taccgccgca cgcacctgag cactgcaaac gggccgagcg
     4921 cctggtgctg ctcgtccggg gactgcagct cctatcgtcc gggatgaatc tggcttccca
     4981 gcagctaagc aatggccagc taaagccgtc gtccaatgtg aagaatgctt tgctcaccat
     5041 gaacgccaaa taccggagca tgctgttcga atcgaaacgc ctcaatggca gcggactatt
     5101 gcagaaggcg aatgcattta acataacggc ggacaagatt ctctatgact acgcactgga
     5161 tatgtgccag gccgctgctc tggatgagct actgaagaac acgaagaatt gcttcgaacg
     5221 ttacaacacg gcacacattc tgcttcattc gttggtgcaa aagtgcaatc atccgcagga
     5281 caaaatgatg ctgaacaaat atcgcgatgc cgttgagaag cgtttgagca tattgcagca
     5341 gcatggctat atatacatga ccgatgagaa tgcttagctc gagcggccgc atcgtgactg
     5401 actgacgatc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     5461 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     5521 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     5581 cggagtgtat aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5641 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5701 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5761 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5821 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5881 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5941 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     6001 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     6061 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     6121 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6181 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     6241 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6301 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     6361 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6421 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6481 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6541 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6601 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6661 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6721 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6781 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6841 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6901 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6961 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     7021 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     7081 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     7141 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7201 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     7261 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7321 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7381 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7441 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     7501 tgattctgtg gataaccgta ttaccgcc
//
