LOCUS       sequence-273236-        6742 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 6742)
  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 6742)
  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..6742
                     /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             2999..3025
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      3042..3051
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3055..3189
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3092..3111)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3146..3165
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3258..3615
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3258..3449
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3281..3301)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3466..3601
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3635..3652)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3635..3651)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3644..3666)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3864..4319
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3951..3970)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4161..4182
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4336..4355)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4455..4477
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4515..4533)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4601..4705
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4706..4774,4775..5566)
                     /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(4924..4943)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5737..6325
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6226..6245
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6479..6496
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6613..6634
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6649..6654,6655..6672,6673..6679)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6687..6703
                     /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     6692..6714
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6711..6727
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6711..6727
                     /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 ccactcccag gtccaactgt
      901 acctcgcgag ccagccatgg aagaatcacc actaagccgg gcaccatccc gtggtggagt
      961 caactttctc aatgtagccc ggacctacat ccccaacacc aaggtggaat gtcactacac
     1021 ccttccccca ggcaccatgc ccagtgccag tgactggatt ggcatcttca aggtggaggc
     1081 tgcctgtgtt cgggattacc acacatttgt gtggtcttcc gtgcctgaaa gtacaactga
     1141 tggttccccc attcacacca gtgtccagtt ccaagccagc tacctgccca aaccaggagc
     1201 tcagctctac cagttccgat atgtgaaccg ccagggccag gtgtgtgggc agagcccccc
     1261 tttccagttc cgagagccaa ggcccatgga tgaactggtg accctggagg aggctgatgg
     1321 gggctctgac atcctgctgg ttgtccccaa ggcaactgtg ttacagaacc agctcgatga
     1381 gagccagcaa gaacggaatg acctgatgca gctgaagcta cagctggagg gacaggtgac
     1441 agagctgagg agccgagtgc aggagctcga gagggctctg gcaactgcca ggcaggagca
     1501 cacggagctg atggaacagt acaaggggat ttcccggtcc catggggaga tcacagaaga
     1561 gagggacatc ctgagccggc aacagggaga ccatgtggca cgcatcctgg agctagagga
     1621 tgacatccag accatcagtg agaaagtgct gacgaaggaa gtggagctgg acaggcttag
     1681 agacacagtg aaggccctga ctcgggaaca agagaagctc cttgggcaac tgaaagaagt
     1741 acaagcagac aaggagcaaa gtgaggctga gctccaagtg gcacaacagg agaaccatca
     1801 cttaaatttg gacctgaagg aggcgaagag ctggcaagag gagcagagtg ctcaggctca
     1861 gcgactgaaa gacaaggtgg cccagatgaa ggacacccta ggccaggccc agcagcgggt
     1921 ggccgagctg gagcccttga aggagcagct tcgaggggcc caggagcttg cagcctcaag
     1981 ccagcagaaa gccacccttc ttggggagga gttggccagt gcagcagcag ccagggaccg
     2041 caccatagcc gaactacacc gcagccgcct ggaagtggct gaagttaacg gcaggctggc
     2101 tgagctcggt ttgcacttga aggaagaaaa atgccaatgg agcaaggagc gggcagggct
     2161 gctgcagagt gtggaggcag agaaggacaa gatcctgaag ctgagtgcag agatacttcg
     2221 attggagaag gcagttcagg aggagaggac ccaaaaccaa gtgttcaaga ctgagctggc
     2281 ccgggagaag gattctagcc tggtacagtt gtcagaaagt aagcgggagc tgacagagct
     2341 gcggtcagcc ctgcgtgtgc tccagaagga aaaggagcag ttacaggagg agaaacagga
     2401 attgctagag tacatgagaa agctagaggc ccgcctggag aaggtggcag atgagaagtg
     2461 gaatgaggat gccaccacag aggatgagga ggccgctgtg gggctgagct gcccggcagc
     2521 tctgacagac tcagaggacg agtccccaga agacatgagg ctcccaccct atggcctttg
     2581 tgagcgtgga gacccaggct cctctcctgc tgggcctcga gaggcttctc cccttgttgt
     2641 catcagccag ccggctccca tttctcctca cctctctggg ccagctgagg acagtagctc
     2701 tgactcggag gctgaagatg agaagtcagt cctgatggca gctgtgcaga gtgggggtga
     2761 ggaggccaac ttactgcttc ctgaactggg cagtgccttc tatgacatgg ccagtggctt
     2821 tacagtgggt accctgtcag aaaccagcac tgggggccct gccaccccca catggaagga
     2881 gtgtcctatc tgtaaggagc gctttcctgc tgagagtgac aaggatgccc tggaggacca
     2941 catggatgga cacttctttt tcagcaccca ggaccccttc acctttgagt ctagaatgta
     3001 cccatacgac gtcccagact acgctggtac ctagaagctt ggccgccatg gcccaacttg
     3061 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3121 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3181 gtctggatcg ggaattaatt cggcgcagca ccatggcctg aaataacctc tgaaagagga
     3241 acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     3301 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3361 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3421 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3481 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3541 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3601 ctaggctttt gcaaaaagct gttaacagct tggcactggc cgtcgtttta caacgtcgtg
     3661 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     3721 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     3781 atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3841 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3901 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3961 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4021 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     4081 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4141 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4201 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4261 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     4321 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4381 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4441 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4501 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4561 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4621 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4681 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4741 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4801 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4861 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4921 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4981 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5041 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5101 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5161 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5221 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5281 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5341 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5401 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5461 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5521 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5581 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5641 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5701 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5761 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5821 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5881 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5941 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6001 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6061 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6121 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6181 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6241 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6301 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6361 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6421 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     6481 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     6541 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     6601 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     6661 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     6721 ctatgacatg attacgaatt aa
//
