LOCUS       sequence-329420-        6479 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis tagged ATG13 [571–700] fragment with 14 scramble 
            amino acid sequence DIKLIDTVDLESCN under a Cu promoter in yeast..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6479)
  AUTHORS   Gatica D, Damasio A, Pascual C, Klionsky DJ, Ragusa MJ, Popelka H
  TITLE     The carboxy terminus of yeast Atg13 binds phospholipid membrane via 
            motifs that overlap with the Vac8-interacting domain.
  JOURNAL   Autophagy. 2020 Jun;16(6):1007-1020. doi: 
            10.1080/15548627.2019.1648117. Epub 2019 Aug 2.
  PUBMED    31352862
REFERENCE   2  (bases 1 to 6479)
  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: 
            "10.1080/15548627.2019.1648117"; journalName: "Autophagy"; date: 
            "2020-06"; volume: "16"; issue: "6"; pages: "1007-1020"
COMMENT     Sequence Label: pCu-His6-Atg13[571–700]-Ss(424)
FEATURES             Location/Qualifiers
     source          1..6479
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     381..398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    515..536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(551..556,557..574,575..581)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    589..605
                     /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     594..616
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     613..629
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     613..629
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     648..668
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        650..668
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        687..921
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             945..962
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1370..1386)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1371..1387)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      1389..1636
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1405..1423)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1654..1673)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1655..1673)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1683..1700)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1683..1699)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1692..1714)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1840..2295
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1927..1946)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2137..2158
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2398..3072)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(3073..3354)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3375..3394)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3494..3516
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3554..3572)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      3613..4955
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    3945..3992
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        4982..5086
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5087..5155,5156..5947)
                     /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     complement(5305..5324)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6118..6479,1..227)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
       61 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
      241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
      301 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
      361 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
      421 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
      481 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      541 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      601 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
      661 actaaaggga acaaaagctg gagctcctag ttagaaaaag acatttttgc tgtcagtcac
      721 tgtcaagaga ttcttttgct ggcatttctt ctagaagcaa aaagagcgat gcgtcttttc
      781 cgctgaaccg ttccagcaaa aaagactacc aacgcaatat ggattgtcag aatcatataa
      841 aagagaagca aataactcct tgtcttgtat caattgcatt ataatatctt cttgttagtg
      901 caatatcata tagaagtcat cgactagtgg atccccttaa gatgcaccat caccatcatc
      961 actctactag tgccttgaat agtagaagga attctttaga taagagctca aacaagcagg
     1021 gtatgtcagg cttacctcct atttttggtg gagagagtac ttcatatcac cacgacaaca
     1081 aaatacaaaa gtacaaccaa ttaggagtag aagaagatga tgatgacgag aatgaccgtt
     1141 tgctcaacca aatgggaaac agtgctacaa aattcaaaag ttcaatatct ccaagatcaa
     1201 ttgatagcat ttcaagttct ttcataaaaa gtaggatacc tatcagacaa ccataccatt
     1261 actctcaacc aactactgcg ccctttcaag ctcaggcgaa atttcataaa cctgcaaata
     1321 agttaatcga taatggtaat aggagtaata gttaggatat caagcttatc gataccgtcg
     1381 acctcgagtc atgtaattag ttatgtcacg cttacattca cgccctcccc ccacatccgc
     1441 tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat
     1501 agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacag
     1561 acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc
     1621 gaaggcttta atttgcggcc ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc
     1681 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     1741 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     1801 cgcccttccc aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat
     1861 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     1921 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     1981 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2041 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2101 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2161 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     2221 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     2281 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     2341 ccgcatagat cggcaagtgc acaaacaata cttaaataaa tactactcag taataaccta
     2401 tttcttagca tttttgacga aatttgctat tttgttagag tcttttacac catttgtctc
     2461 cacacctccg cttacatcaa caccaataac gccatttaat ctaagcgcat caccaacatt
     2521 ttctggcgtc agtccaccag ctaacataaa atgtaagctt tcggggctct cttgccttcc
     2581 aacccagtca gaaatcgagt tccaatccaa aagttcacct gtcccacctg cttctgaatc
     2641 aaacaaggga ataaacgaat gaggtttctg tgaagctgca ctgagtagta tgttgcagtc
     2701 ttttggaaat acgagtcttt taataactgg caaaccgagg aactcttggt attcttgcca
     2761 cgactcatct ccatgcagtt ggacgatatc aatgccgtaa tcattgacca gagccaaaac
     2821 atcctcctta ggttgattac gaaacacgcc aaccaagtat ttcggagtgc ctgaactatt
     2881 tttatatgct tttacaagac ttgaaatttt ccttgcaata accgggtcaa ttgttctctt
     2941 tctattgggc acacatataa tacccagcaa gtcagcatcg gaatctagag cacattctgc
     3001 ggcctctgtg ctctgcaagc cgcaaacttt caccaatgga ccagaactac ctgtgaaatt
     3061 aataacagac atactccaag ctgcctttgt gtgcttaatc acgtatactc acgtgctcaa
     3121 tagtcaccaa tgccctccct cttggccctc tccttttctt ttttcgaccg aattaattct
     3181 taatcggcaa aaaaagaaaa gctccggatc aagattgtac gtaaggtgac aagctatttt
     3241 tcaataaaga atatcttcca ctactgccat ctggcgtcat aactgcaaag tacacatata
     3301 ttacgatgct gttctattaa atgcttccta tattatatat atagtaatgt cgttgatcta
     3361 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     3421 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     3481 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     3541 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     3601 gtttcttagt atgatccaat atcaaaggaa atgatagcat tgaaggatga gactaatcca
     3661 attgaggagt ggcagcatat agaacagcta aagggtagtg ctgaaggaag catacgatac
     3721 cccgcatgga atgggataat atcacaggag gtactagact acctttcatc ctacataaat
     3781 agacgcatat aagtacgcat ttaagcataa acacgcacta tgccgttctt ctcatgtata
     3841 tatatataca ggcaacacgc agatataggt gcgacgtgaa cagtgagctg tatgtgcgca
     3901 gctcgcgttg cattttcgga agcgctcgtt ttcggaaacg ctttgaagtt cctattccga
     3961 agttcctatt ctctagaaag tataggaact tcagagcgct tttgaaaacc aaaagcgctc
     4021 tgaagacgca ctttcaaaaa accaaaaacg caccggactg taacgagcta ctaaaatatt
     4081 gcgaataccg cttccacaaa cattgctcaa aagtatctct ttgctatata tctctgtgct
     4141 atatccctat ataacctacc catccacctt tcgctccttg aacttgcatc taaactcgac
     4201 ctctacattt tttatgttta tctctagtat tactctttag acaaaaaaat tgtagtaaga
     4261 actattcata gagtgaatcg aaaacaatac gaaaatgtaa acatttccta tacgtagtat
     4321 atagagacaa aatagaagaa accgttcata attttctgac caatgaagaa tcatcaacgc
     4381 tatcactttc tgttcacaaa gtatgcgcaa tccacatcgg tatagaatat aatcggggat
     4441 gcctttatct tgaaaaaatg cacccgcagc ttcgctagta atcagtaaac gcgggaagtg
     4501 gagtcaggct ttttttatgg aagagaaaat agacaccaaa gtagccttct tctaacctta
     4561 acggacctac agtgcaaaaa gttatcaaga gactgcatta tagagcgcac aaaggagaaa
     4621 aaaagtaatc taagatgctt tgttagaaaa atagcgctct cgggatgcat ttttgtagaa
     4681 caaaaaagaa gtatagattc tttgttggta aaatagcgct ctcgcgttgc atttctgttc
     4741 tgtaaaaatg cagctcagat tctttgtttg aaaaattagc gctctcgcgt tgcatttttg
     4801 ttttacaaaa atgaagcaca gattcttcgt tggtaaaata gcgctttcgc gttgcatttc
     4861 tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat
     4921 ttttgttcta caaaatgaag cacagatgct tcgttcaggt ggcacttttc ggggaaatgt
     4981 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5041 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5101 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5161 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5221 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5281 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5341 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5401 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5461 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5521 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5581 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5641 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5701 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5761 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5821 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5881 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5941 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6001 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6061 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6121 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6181 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6241 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6301 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6361 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6421 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgacct
//
