LOCUS       sequence-266001-        6955 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 6955)
  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 6955)
  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..6955
                     /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             3121..3150
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3169..3192
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3212..3228)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3212..3228)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3239..3861
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3438..3457)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(4010..4598)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4090..4109)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4927..4974
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4999..5018
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5206..6000)
                     /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(5206..6000)
                     /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(5317..5336)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5927..5946
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6035..6392)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6049..6184
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6201..6392
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6349..6369
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(6394..6498)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6525..6953
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6612..6631)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6822..6843
                     /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 tggatccggt accgaggaga tctgccgccg
     1021 cgatcgccca tggaagaatc accactaagc cgggcaccat cccgtggtgg agtcaacttt
     1081 ctcaatgtag cccggaccta catccccaac accaaggtgg aatgtcacta cacccttccc
     1141 ccaggcacca tgcccagtgc cagtgactgg attggcatct tcaaggtgga ggctgcctgt
     1201 gttcgggatt accacacatt tgtgtggtct tccgtgcctg aaagtacaac tgatggttcc
     1261 cccattcaca ccagtgtcca gttccaagcc agctacctgc ccaaaccagg agctcagctc
     1321 taccagttcc gatatgtgaa ccgccagggc caggtgtgtg ggcagagccc ccctttccag
     1381 ttccgagagc caaggcccat ggatgaactg gtgaccctgg aggaggctga tgggggctct
     1441 gacatcgcgc tggttgtccc caaggcaact gtgttacaga accagctcga tgagagccag
     1501 caagaacgga atgacctgat gcagctgaag ctacagctgg agggacaggt gacagagctg
     1561 aggagccgag tgcaggagct cgagagggct ctggcaactg ccaggcagga gcacacggag
     1621 ctgatggaac agtacaaggg gatttcccgg tcccatgggg agatcacaga agagagggac
     1681 atcctgagcc ggcaacaggg agaccatgtg gcacgcatcc tggagctaga ggatgacatc
     1741 cagaccatca gtgagaaagt gctgacgaag gaagtggagc tggacaggct tagagacaca
     1801 gtgaaggccc tgactcggga acaagagaag ctccttgggc aactgaaaga agtacaagca
     1861 gacaaggagc aaagtgaggc tgagctccaa gtggcacaac aggagaacca tcacttaaat
     1921 ttggacctga aggaggcgaa gagctggcaa gaggagcaga gtgctcaggc tcagcgactg
     1981 aaagacaagg tggcccagat gaaggacacc ctaggccagg cccagcagcg ggtggccgag
     2041 ctggagccct tgaaggagca gcttcgaggg gcccaggagc ttgcagcctc aagccagcag
     2101 aaagccaccc ttcttgggga ggagttggcc agtgcagcag cagccaggga ccgcaccata
     2161 gccgaactac accgcagccg cctggaagtg gctgaagtta acggcaggct ggctgagctc
     2221 ggtttgcact tgaaggaaga aaaatgccaa tggagcaagg agcgggcagg gctgctgcag
     2281 agtgtggagg cagagaagga caagatcctg aagctgagtg cagagatact tcgattggag
     2341 aaggcagttc aggaggagag gacccaaaac caagtgttca agactgagct ggcccgggag
     2401 aaggattcta gcctggtaca gttgtcagaa agtaagcggg agctgacaga gctgcggtca
     2461 gccctgcgtg tgctccagaa ggaaaaggag cagttacagg aggagaaaca ggaattgcta
     2521 gagtacatga gaaagctaga ggcccgcctg gagaaggtgg cagatgagaa gtggaatgag
     2581 gatgccacca cagaggatga ggaggccgct gtggggctga gctgcccggc agctctgaca
     2641 gactcagagg acgagtcccc agaagacatg aggctcccac cctatggcct ttgtgagcgt
     2701 ggagacccag gctcctctcc tgctgggcct cgagaggctt ctccccttgt tgtcatcagc
     2761 cagccggctc ccatttctcc tcacctctct gggccagctg aggacagtag ctctgactcg
     2821 gaggctgaag atgagaagtc agtcctgatg gcagctgtgc agagtggggg tgaggaggcc
     2881 aacttactgc ttcctgaact gggcagtgcc ttctatgaca tggccagtgg ctttacagtg
     2941 ggtaccctgt cagaaaccag cactgggggc cctgccaccc ccacatggaa ggagtgtcct
     3001 atctgtaagg agcgctttcc tgctgagagt gacaaggatg ccctggagga ccacatggat
     3061 ggacacttct ttttcagcac ccaggacccc ttcacctttg agacgcgtac gcggccgctc
     3121 gagcagaaac tcatctcaga agaggatctg gcagcaaatg atatcctgga ttacaaggat
     3181 gacgacgata aggtttaaac ggccggccgc ggtcatagct gtttcctgaa cagatcccgg
     3241 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt
     3301 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt
     3361 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac
     3421 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct
     3481 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt
     3541 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg
     3601 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg
     3661 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat
     3721 tttaaaataa ctataccagc aggaggacgt ccagacacag cataggctac ctggccatgc
     3781 ccaaccggtg ggacatttga gttgcttgct tggcactgtc ctctcatgcg ttgggtccac
     3841 tcagtagatg cctgttgaat tgggtacgcg gccagcggcg agcggtatca gctcactcaa
     3901 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3961 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4021 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4081 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4141 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4201 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4261 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4321 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4381 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4441 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4501 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4561 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4621 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4681 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4741 gtatatatga gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc
     4801 ctgggccttc acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt
     4861 ggccccaatg gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc
     4921 gtttatgaac aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag
     4981 cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg
     5041 cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa
     5101 cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca
     5161 ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc
     5221 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag
     5281 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat
     5341 gtcctgatag cgatccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc
     5401 attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc
     5461 gtcgggcatg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc
     5521 ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat
     5581 gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg
     5641 cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc
     5701 ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag
     5761 cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg
     5821 cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc
     5881 tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc
     5941 gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat
     6001 gcgaaacgat cctcatcctg tctcttgatc gatctttgca aaagcctagg cctccaaaaa
     6061 agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa
     6121 taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg
     6181 ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca
     6241 tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac taattgagat
     6301 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac
     6361 acacattcca cagctggttc tttccgcctc aggactcttc ctttttcaat attattgaag
     6421 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6481 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg
     6541 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     6601 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     6661 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     6721 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     6781 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     6841 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     6901 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga atttt
//
