LOCUS       sequence-292247-        6953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CALCOCO1 CLIR 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 L140A-mycDDK
FEATURES             Location/Qualifiers
     source          1..6953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     152..174
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     166..183
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     167..183
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        344..723
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        724..927
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     878..898
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     953..972
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        953..971
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3120..3149
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3168..3191
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3211..3227)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3211..3227)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3238..3860
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3437..3456)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(4009..4597)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4089..4108)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4926..4973
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4998..5017
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5205..5999)
                     /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(5205..5999)
                     /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(5316..5335)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5926..5945
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6034..6391)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6048..6183
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6200..6391
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6348..6368
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(6393..6497)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6524..6952
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6611..6630)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6821..6842
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 taacaaaata ttaacgctta caatttccat tcgccattca ggctgcgcaa ctgttgggaa
       61 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
      121 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
      181 agtgccaagc tgatctatac attgaatcaa tattggcaat tagccatatt agtcattggt
      241 tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat
      301 atgtacattt atattggctc atgtccaata tgaccgccat gttgacattg attattgact
      361 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      421 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      481 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      541 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      601 agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      661 atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      721 atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga
      781 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      841 gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta
      901 cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagaattt tgtaatacga
      961 ctcactatag ggcggccggg aattcgtcga ctggatccag taccgaggag atctgcgccg
     1021 cgatcgccat ggaagaatca ccactaagcc gggcaccatc ccgtggtgga gtcaactttc
     1081 tcaatgtagc ccggacctac atccccaaca ccaaggtgga atgtcactac acccttcccc
     1141 caggcaccat gcccagtgcc agtgactgga ttggcatctt caaggtggag gctgcctgtg
     1201 ttcgggatta ccacacattt gtgtggtctt ccgtgcctga aagtacaact gatggttccc
     1261 ccattcacac cagtgtccag ttccaagcca gctacctgcc caaaccagga gctcagctct
     1321 accagttccg atatgtgaac cgccagggcc aggtgtgtgg gcagagcccc cctttccagt
     1381 tccgagagcc aaggcccatg gatgaactgg tgaccctgga ggaggctgat gggggctctg
     1441 acatcgcgct ggttgtcccc aaggcaactg tgttacagaa ccagctcgat gagagccagc
     1501 aagaacggaa tgacctgatg cagctgaagc tacagctgga gggacaggtg acagagctga
     1561 ggagccgagt gcaggagctc gagagggctc tggcaactgc caggcaggag cacacggagc
     1621 tgatggaaca gtacaagggg atttcccggt cccatgggga gatcacagaa gagagggaca
     1681 tcctgagccg gcaacaggga gaccatgtgg cacgcatcct ggagctagag gatgacatcc
     1741 agaccatcag tgagaaagtg ctgacgaagg aagtggagct ggacaggctt agagacacag
     1801 tgaaggccct gactcgggaa caagagaagc tccttgggca actgaaagaa gtacaagcag
     1861 acaaggagca aagtgaggct gagctccaag tggcacaaca ggagaaccat cacttaaatt
     1921 tggacctgaa ggaggcgaag agctggcaag aggagcagag tgctcaggct cagcgactga
     1981 aagacaaggt ggcccagatg aaggacaccc taggccaggc ccagcagcgg gtggccgagc
     2041 tggagccctt gaaggagcag cttcgagggg cccaggagct tgcagcctca agccagcaga
     2101 aagccaccct tcttggggag gagttggcca gtgcagcagc agccagggac cgcaccatag
     2161 ccgaactaca ccgcagccgc ctggaagtgg ctgaagttaa cggcaggctg gctgagctcg
     2221 gtttgcactt gaaggaagaa aaatgccaat ggagcaagga gcgggcaggg ctgctgcaga
     2281 gtgtggaggc agagaaggac aagatcctga agctgagtgc agagatactt cgattggaga
     2341 aggcagttca ggaggagagg acccaaaacc aagtgttcaa gactgagctg gcccgggaga
     2401 aggattctag cctggtacag ttgtcagaaa gtaagcggga gctgacagag ctgcggtcag
     2461 ccctgcgtgt gctccagaag gaaaaggagc agttacagga ggagaaacag gaattgctag
     2521 agtacatgag aaagctagag gcccgcctgg agaaggtggc agatgagaag tggaatgagg
     2581 atgccaccac agaggatgag gaggccgctg tggggctgag ctgcccggca gctctgacag
     2641 actcagagga cgagtcccca gaagacatga ggctcccacc ctatggcctt tgtgagcgtg
     2701 gagacccagg ctcctctcct gctgggcctc gagaggcttc tccccttgtt gtcatcagcc
     2761 agccggctcc catttctcct cacctctctg ggccagctga ggacagtagc tctgactcgg
     2821 aggctgaaga tgagaagtca gtcctgatgg cagctgtgca gagtgggggt gaggaggcca
     2881 acttactgct tcctgaactg ggcagtgcct tctatgacat ggccagtggc tttacagtgg
     2941 gtaccctgtc agaaaccagc actgggggcc ctgccacccc cacatggaag gagtgtccta
     3001 tctgtaagga gcgctttcct gctgagagtg acaaggatgc cctggaggac cacatggatg
     3061 gacacttctt tttcagcacc caggacccct tcacctttga gacgcgtacg cggccgctcg
     3121 agcagaaact catctcagaa gaggatctgg cagcaaatga tatcctggat tacaaggatg
     3181 acgacgataa ggtttaaacg gccggccgcg gtcatagctg tttcctgaac agatcccggg
     3241 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     3301 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     3361 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     3421 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     3481 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
     3541 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     3601 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     3661 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
     3721 ttaaaataac tataccagca ggaggacgtc cagacacagc ataggctacc tggccatgcc
     3781 caaccggtgg gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact
     3841 cagtagatgc ctgttgaatt gggtacgcgg ccagcggcga gcggtatcag ctcactcaaa
     3901 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3961 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4021 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4081 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4141 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4201 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4261 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4321 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4381 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4441 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4501 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4561 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4621 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4681 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4741 tatatatgag taacctgagg ctatggcagg gcctgccgcc ccgacgttgg ctgcgagccc
     4801 tgggccttca cccgaacttg gggggtgggg tggggaaaag gaagaaacgc gggcgtattg
     4861 gccccaatgg ggtctcggtg gggtatcgac agagtgccag ccctgggacc gaaccccgcg
     4921 tttatgaaca aacgacccaa caccgtgcgt tttattctgt ctttttattg ccgtcatagc
     4981 gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc tagggtgggc
     5041 gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac
     5101 gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag
     5161 gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca
     5221 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
     5281 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
     5341 tcctgatagc gatccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
     5401 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
     5461 tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
     5521 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
     5581 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
     5641 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     5701 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     5761 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc
     5821 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
     5881 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
     5941 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg
     6001 cgaaacgatc ctcatcctgt ctcttgatcg atctttgcaa aagcctaggc ctccaaaaaa
     6061 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat
     6121 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
     6181 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
     6241 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
     6301 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
     6361 cacattccac agctggttct ttccgcctca ggactcttcc tttttcaata ttattgaagc
     6421 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6481 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc
     6541 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     6601 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     6661 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     6721 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     6781 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     6841 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     6901 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttt
//
