LOCUS       sequence-273245-        6953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CALCOCO1 R12H mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6953)
  AUTHORS   Stefely JA, Zhang Y, Freiberger EC, Kwiecien NW, Thomas HE, Davis 
            AM, Lowry ND, Vincent CE, Shishkova E, Clark NA, Medvedovic M, Coon 
            JJ, Pagliarini DJ, Mercer CA
  TITLE     Mass spectrometry proteomics reveals a function for mammalian 
            CALCOCO1 in MTOR-regulated selective autophagy.
  JOURNAL   Autophagy. 2020 Jan 23. doi: 10.1080/15548627.2020.1719746.
  PUBMED    31971854
REFERENCE   2  (bases 1 to 6953)
  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"; journalName: "Autophagy. 
            2020 Jan 23. doi: 10.1080/15548627.2020.1719746."
COMMENT     Sequence Label: CALCOCO1 R12H-mycDDK
FEATURES             Location/Qualifiers
     source          1..6953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..173
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     165..182
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     877..897
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     952..971
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3119..3148
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3167..3190
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3210..3226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3210..3226)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3237..3859
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3436..3455)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(4008..4596)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4088..4107)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4925..4972
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4997..5016
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5204..5998)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(5204..5998)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5315..5334)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5925..5944
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6033..6390)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6047..6182
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6199..6390
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6347..6367
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(6392..6496)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6523..6951
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6610..6629)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6820..6841
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccagt accgaggaga tctgcgccgc
     1021 gatcgccatg gaagaatcac cactaagccg ggcaccatcc catggtggag tcaactttct
     1081 caatgtagcc cggacctaca tccccaacac caaggtggaa tgtcactaca cccttccccc
     1141 aggcaccatg cccagtgcca gtgactggat tggcatcttc aaggtggagg ctgcctgtgt
     1201 tcgggattac cacacatttg tgtggtcttc cgtgcctgaa agtacaactg atggttcccc
     1261 cattcacacc agtgtccagt tccaagccag ctacctgccc aaaccaggag ctcagctcta
     1321 ccagttccga tatgtgaacc gccagggcca ggtgtgtggg cagagccccc ctttccagtt
     1381 ccgagagcca aggcccatgg atgaactggt gaccctggag gaggctgatg ggggctctga
     1441 catcctgctg gttgtcccca aggcaactgt gttacagaac cagctcgatg agagccagca
     1501 agaacggaat gacctgatgc agctgaagct acagctggag ggacaggtga cagagctgag
     1561 gagccgagtg caggagctcg agagggctct ggcaactgcc aggcaggagc acacggagct
     1621 gatggaacag tacaagggga tttcccggtc ccatggggag atcacagaag agagggacat
     1681 cctgagccgg caacagggag accatgtggc acgcatcctg gagctagagg atgacatcca
     1741 gaccatcagt gagaaagtgc tgacgaagga agtggagctg gacaggctta gagacacagt
     1801 gaaggccctg actcgggaac aagagaagct ccttgggcaa ctgaaagaag tacaagcaga
     1861 caaggagcaa agtgaggctg agctccaagt ggcacaacag gagaaccatc acttaaattt
     1921 ggacctgaag gaggcgaaga gctggcaaga ggagcagagt gctcaggctc agcgactgaa
     1981 agacaaggtg gcccagatga aggacaccct aggccaggcc cagcagcggg tggccgagct
     2041 ggagcccttg aaggagcagc ttcgaggggc ccaggagctt gcagcctcaa gccagcagaa
     2101 agccaccctt cttggggagg agttggccag tgcagcagca gccagggacc gcaccatagc
     2161 cgaactacac cgcagccgcc tggaagtggc tgaagttaac ggcaggctgg ctgagctcgg
     2221 tttgcacttg aaggaagaaa aatgccaatg gagcaaggag cgggcagggc tgctgcagag
     2281 tgtggaggca gagaaggaca agatcctgaa gctgagtgca gagatacttc gattggagaa
     2341 ggcagttcag gaggagagga cccaaaacca agtgttcaag actgagctgg cccgggagaa
     2401 ggattctagc ctggtacagt tgtcagaaag taagcgggag ctgacagagc tgcggtcagc
     2461 cctgcgtgtg ctccagaagg aaaaggagca gttacaggag gagaaacagg aattgctaga
     2521 gtacatgaga aagctagagg cccgcctgga gaaggtggca gatgagaagt ggaatgagga
     2581 tgccaccaca gaggatgagg aggccgctgt ggggctgagc tgcccggcag ctctgacaga
     2641 ctcagaggac gagtccccag aagacatgag gctcccaccc tatggccttt gtgagcgtgg
     2701 agacccaggc tcctctcctg ctgggcctcg agaggcttct ccccttgttg tcatcagcca
     2761 gccggctccc atttctcctc acctctctgg gccagctgag gacagtagct ctgactcgga
     2821 ggctgaagat gagaagtcag tcctgatggc agctgtgcag agtgggggtg aggaggccaa
     2881 cttactgctt cctgaactgg gcagtgcctt ctatgacatg gccagtggct ttacagtggg
     2941 taccctgtca gaaaccagca ctgggggccc tgccaccccc acatggaagg agtgtcctat
     3001 ctgtaaggag cgctttcctg ctgagagtga caaggatgcc ctggaggacc acatggatgg
     3061 acacttcttt ttcagcaccc aggacccctt cacctttgag acgcgtacgc ggccgctcga
     3121 gcagaaactc atctcagaag aggatctggc agcaaatgat atcctggatt acaaggatga
     3181 cgacgataag gtttaaacgg ccggccgcgg tcatagctgt ttcctgaaca gatcccgggt
     3241 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
     3301 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
     3361 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
     3421 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
     3481 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
     3541 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
     3601 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
     3661 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
     3721 taaaataact ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc
     3781 aaccggtggg acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc
     3841 agtagatgcc tgttgaattg ggtacgcggc cagcggcgag cggtatcagc tcactcaaag
     3901 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3961 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4021 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4081 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4141 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4201 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4261 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4321 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4381 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4441 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4501 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4561 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4621 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4681 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4741 atatatgagt aacctgaggc tatggcaggg cctgccgccc cgacgttggc tgcgagccct
     4801 gggccttcac ccgaacttgg ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg
     4861 ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt
     4921 ttatgaacaa acgacccaac accgtgcgtt ttattctgtc tttttattgc cgtcatagcg
     4981 cgggttcctt ccggtattgt ctccttccgt gtttcagtta gcctccccct agggtgggcg
     5041 aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg
     5101 attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg
     5161 ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa
     5221 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     5281 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     5341 cctgatagcg atccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     5401 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
     5461 cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     5521 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
     5581 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
     5641 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
     5701 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
     5761 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca
     5821 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
     5881 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
     5941 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc
     6001 gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa agcctaggcc tccaaaaaag
     6061 cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata
     6121 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
     6181 atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata
     6241 cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc
     6301 atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac
     6361 acattccaca gctggttctt tccgcctcag gactcttcct ttttcaatat tattgaagca
     6421 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6481 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
     6541 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     6601 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     6661 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     6721 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     6781 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     6841 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     6901 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat ttt
//
