LOCUS       sequence-273225-        7477 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 7477)
  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 7477)
  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: GFP-CALCOCO1
FEATURES             Location/Qualifiers
     source          1..7477
                     /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             join(926..928,929..931,932..1642)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(926..928,929..931,932..1642)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(971..992)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1232..1251)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1579..1600
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3737..3766
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      3777..3786
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3790..3924
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3827..3846)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3881..3900
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3993..4350
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3993..4184
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4016..4036)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4201..4336
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4370..4387)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4370..4386)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4379..4401)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4599..5054
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4686..4705)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4896..4917
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5071..5090)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5190..5212
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5250..5268)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5336..5440
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5441..5509,5510..6301)
                     /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(5659..5678)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6472..7060
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6961..6980
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7214..7231
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7348..7369
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7384..7389,7390..7407,7408..7414)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7422..7438
                     /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     7427..7449
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7446..7462
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7446..7462
                     /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 gtccaactgc
      901 acctcggttc tatcgcgagc cagccatggt gagcaagggc gaggagctgt tcaccggggt
      961 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     1021 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     1081 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     1141 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     1201 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     1261 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     1321 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     1381 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     1441 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     1501 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     1561 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     1621 cggcatggac gagctgtaca agggcggccg catggaagaa tcaccactaa gccgggcacc
     1681 atcccgtggt ggagtcaact ttctcaatgt agcccggacc tacatcccca acaccaaggt
     1741 ggaatgtcac tacacccttc ccccaggcac catgcccagt gccagtgact ggattggcat
     1801 cttcaaggtg gaggctgcct gtgttcggga ttaccacaca tttgtgtggt cttccgtgcc
     1861 tgaaagtaca actgatggtt cccccattca caccagtgtc cagttccaag ccagctacct
     1921 gcccaaacca ggagctcagc tctaccagtt ccgatatgtg aaccgccagg gccaggtgtg
     1981 tgggcagagc ccccctttcc agttccgaga gccaaggccc atggatgaac tggtgaccct
     2041 ggaggaggct gatgggggct ctgacatcct gctggttgtc cccaaggcaa ctgtgttaca
     2101 gaaccagctc gatgagagcc agcaagaacg gaatgacctg atgcagctga agctacagct
     2161 ggagggacag gtgacagagc tgaggagccg agtgcaggag ctcgagaggg ctctggcaac
     2221 tgccaggcag gagcacacgg agctgatgga acagtacaag gggatttccc ggtcccatgg
     2281 ggagatcaca gaagagaggg acatcctgag ccggcaacag ggagaccatg tggcacgcat
     2341 cctggagcta gaggatgaca tccagaccat cagtgagaaa gtgctgacga aggaagtgga
     2401 gctggacagg cttagagaca cagtgaaggc cctgactcgg gaacaagaga agctccttgg
     2461 gcaactgaaa gaagtacaag cagacaagga gcaaagtgag gctgagctcc aagtggcaca
     2521 acaggagaac catcacttaa atttggacct gaaggaggcg aagagctggc aagaggagca
     2581 gagtgctcag gctcagcgac tgaaagacaa ggtggcccag atgaaggaca ccctaggcca
     2641 ggcccagcag cgggtggccg agctggagcc cttgaaggag cagcttcgag gggcccagga
     2701 gcttgcagcc tcaagccagc agaaagccac ccttcttggg gaggagttgg ccagtgcagc
     2761 agcagccagg gaccgcacca tagccgaact acaccgcagc cgcctggaag tggctgaagt
     2821 taacggcagg ctggctgagc tcggtttgca cttgaaggaa gaaaaatgcc aatggagcaa
     2881 ggagcgggca gggctgctgc agagtgtgga ggcagagaag gacaagatcc tgaagctgag
     2941 tgcagagata cttcgattgg agaaggcagt tcaggaggag aggacccaaa accaagtgtt
     3001 caagactgag ctggcccggg agaaggattc tagcctggta cagttgtcag aaagtaagcg
     3061 ggagctgaca gagctgcggt cagccctgcg tgtgctccag aaggaaaagg agcagttaca
     3121 ggaggagaaa caggaattgc tagagtacat gagaaagcta gaggcccgcc tggagaaggt
     3181 ggcagatgag aagtggaatg aggatgccac cacagaggat gaggaggccg ctgtggggct
     3241 gagctgcccg gcagctctga cagactcaga ggacgagtcc ccagaagaca tgaggctccc
     3301 accctatggc ctttgtgagc gtggagaccc aggctcctct cctgctgggc ctcgagaggc
     3361 ttctcccctt gttgtcatca gccagccggc tcccatttct cctcacctct ctgggccagc
     3421 tgaggacagt agctctgact cggaggctga agatgagaag tcagtcctga tggcagctgt
     3481 gcagagtggg ggtgaggagg ccaacttact gcttcctgaa ctgggcagtg ccttctatga
     3541 catggccagt ggctttacag tgggtaccct gtcagaaacc agcactgggg gccctgccac
     3601 ccccacatgg aaggagtgtc ctatctgtaa ggagcgcttt cctgctgaga gtgacaagga
     3661 tgccctggag gaccacatgg atggacactt ctttttcagc acccaggacc ccttcacctt
     3721 tgagtgatct agaatggagc agaagctgat ctccgaggag gacctgtgaa agcttggccg
     3781 ccatggccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3841 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3901 aatgtatctt atcatgtctg gatcgggaat taattcggcg cagcaccatg gcctgaaata
     3961 acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat
     4021 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4081 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4141 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4201 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4261 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     4321 ggcttttttg gaggcctagg cttttgcaaa aagctgttaa cagcttggca ctggccgtcg
     4381 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     4441 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     4501 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
     4561 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     4621 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4681 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4741 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4801 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4861 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4921 aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc
     4981 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     5041 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5101 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5161 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5221 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5281 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5341 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5401 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5461 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5521 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5581 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5641 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5701 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5761 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5821 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5881 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5941 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6001 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6061 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6121 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6181 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6241 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6301 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6361 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6421 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6481 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6541 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6601 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6661 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6721 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6781 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6841 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6901 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6961 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7021 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7081 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     7141 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     7201 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     7261 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     7321 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     7381 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     7441 tcacacagga aacagctatg acatgattac gaattaa
//
