LOCUS       sequence-266000-        6737 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression and localization of tagged CALCOCO1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6737)
  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 6737)
  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-HA
FEATURES             Location/Qualifiers
     source          1..6737
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3000..3026
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      3037..3046
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3050..3184
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3087..3106)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3141..3160
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3253..3610
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3253..3444
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3276..3296)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3461..3596
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3630..3647)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3630..3646)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3639..3661)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3859..4314
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3946..3965)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4156..4177
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4331..4350)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4450..4472
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4510..4528)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4596..4700
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4701..4769,4770..5561)
                     /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(4919..4938)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5732..6320
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6221..6240
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6474..6491
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6608..6629
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6644..6649,6650..6667,6668..6674)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6682..6698
                     /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     6687..6709
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6706..6722
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6706..6722
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccacttccca ggtccaactg
      901 tacctcgcga gccagccatg gaagaatcac cactaagccg ggcaccatcc cgtggtggag
      961 tcaactttct caatgtagcc cggacctaca tccccaacac caaggtggaa tgtcactaca
     1021 cccttccccc aggcaccatg cccagtgcca gtgactggat tggcatcttc aaggtggagg
     1081 ctgcctgtgt tcgggattac cacacatttg tgtggtcttc cgtgcctgaa agtacaactg
     1141 atggttcccc cattcacacc agtgtccagt tccaagccag ctacctgccc aaaccaggag
     1201 ctcagctcta ccagttccga tatgtgaacc gccagggcca ggtgtgtggg cagagccccc
     1261 ctttccagtt ccgagagcca aggcccatgg atgaactggt gaccctggag gaggctgatg
     1321 ggggctctga catcctgctg gttgtcccca aggcaactgt gttacagaac cagctcgatg
     1381 agagccagca agaacggaat gacctgatgc agctgaagct acagctggag ggacaggtga
     1441 cagagctgag gagccgagtg caggagctcg agagggctct ggcaactgcc aggcaggagc
     1501 acacggagct gatggaacag tacaagggga tttcccggtc ccatggggag atcacagaag
     1561 agagggacat cctgagccgg caacagggag accatgtggc acgcatcctg gagctagagg
     1621 atgacatcca gaccatcagt gagaaagtgc tgacgaagga agtggagctg gacaggctta
     1681 gagacacagt gaaggccctg actcgggaac aagagaagct ccttgggcaa ctgaaagaag
     1741 tacaagcaga caaggagcaa agtgaggctg agctccaagt ggcacaacag gagaaccatc
     1801 acttaaattt ggacctgaag gaggcgaaga gctggcaaga ggagcagagt gctcaggctc
     1861 agcgactgaa agacaaggtg gcccagatga aggacaccct aggccaggcc cagcagcggg
     1921 tggccgagct ggagcccttg aaggagcagc ttcgaggggc ccaggagctt gcagcctcaa
     1981 gccagcagaa agccaccctt cttggggagg agttggccag cgcagcagca gccagggacc
     2041 gcaccatagc cgaactacac cgcagccgcc tggaagtggc tgaagttaac ggcaggctgg
     2101 ctgagctcgg tttgcacttg aaggaagaaa aatgccaatg gagcaaggag cgggcagggc
     2161 tgctgcagag tgtggaggca gagaaggaca agatcctgaa gctgagtgca gagatacttc
     2221 gattggagaa ggcagttcag gaggagagga cccaaaacca agtgttcaag actgagctgg
     2281 cccgggagaa ggattctagc ctggtacagt tgtcagaaag taagcgggag ctgacagagc
     2341 tgcggtcagc cctgcgtgtg ctccagaagg aaaaggagca gttacaggag gagaaacagg
     2401 aattgctaga gtacatgaga aagctagagg cccgcctgga gaaggtggca gatgagaagt
     2461 ggaatgagga tgccaccaca gaggatgagg aggccgctgt ggggctgagc tgcccggcag
     2521 ctctgacaga ctcagaggac gagtccccag aagacatgag gctcccaccc tatggccttt
     2581 gtgagcgtgg agacccaggc tcctctcctg ctgggcctcg agaggcttct ccccttgttg
     2641 tcatcagcca gccggctccc atttctcctc acctctctgg gccagctgag gacagtagct
     2701 ctgactcgga ggctgaagat gagaagtcag tcctgatggc agctgtgcag agtgggggtg
     2761 aggaggccaa cttactgctt cctgaactgg gcagtgcctt ctatgacatg gccagtggct
     2821 ttacagtggg taccctgtca gaaaccagca ctgggggccc tgccaccccc acatggaagg
     2881 agtgtcctat ctgtaaggag cgctttcctg ctgagagtga caaggatgcc ctggaggacc
     2941 acatggatgg acacttcttt ttcagcaccc aggacccctt cacctttgag tctagaatgt
     3001 acccatacga cgtcccagac tacgcttaga agcttggccg ccatggccca acttgtttat
     3061 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3121 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3181 gatcgggaat taattcggcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg
     3241 gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     3301 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3361 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3421 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3481 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3541 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3601 cttttgcaaa aagctgttaa cagcttggca ctggccgtcg ttttacaacg tcgtgactgg
     3661 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     3721 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     3781 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     3841 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3901 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3961 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4021 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     4081 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4141 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4201 gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4261 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     4321 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     4381 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4441 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4501 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     4561 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4621 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4681 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4741 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4801 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4861 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4921 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4981 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5041 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5101 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5161 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5221 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5281 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5341 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5401 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5461 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5521 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5581 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5641 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5701 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5761 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5821 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5881 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5941 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6001 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6061 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6121 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6181 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6241 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6301 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6361 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6421 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     6481 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     6541 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     6601 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     6661 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     6721 acatgattac gaattaa
//
