LOCUS       sequence-265999-        7469 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 7469)
  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 7469)
  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..7469
                     /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(918..920,921..923,924..1634)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(918..920,921..923,924..1634)
                     /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(963..984)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1224..1243)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1571..1592
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3729..3758
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      3769..3778
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3782..3916
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3819..3838)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3873..3892
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3985..4342
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3985..4176
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4008..4028)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4193..4328
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4362..4379)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4362..4378)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4371..4393)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4591..5046
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4678..4697)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4888..4909
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5063..5082)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5182..5204
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5242..5260)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5328..5432
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5433..5501,5502..6293)
                     /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(5651..5670)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6464..7052
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6953..6972
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7206..7223
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7340..7361
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7376..7381,7382..7399,7400..7406)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7414..7430
                     /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     7419..7441
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7438..7454
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7438..7454
                     /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 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
      961 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     1021 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     1081 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     1141 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1201 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1261 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1321 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     1381 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     1441 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1501 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     1561 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1621 acgagctgta caagggcggc cgcatggaag aatcaccact aagccgggca ccatcccgtg
     1681 gtggagtcaa ctttctcaat gtagcccgga cctacatccc caacaccaag gtggaatgtc
     1741 actacaccct tcccccaggc accatgccca gtgccagtga ctggattggc atcttcaagg
     1801 tggaggctgc ctgtgttcgg gattaccaca catttgtgtg gtcttccgtg cctgaaagta
     1861 caactgatgg ttcccccatt cacaccagtg tccagttcca agccagctac ctgcccaaac
     1921 caggagctca gctctaccag ttccgatatg tgaaccgcca gggccaggtg tgtgggcaga
     1981 gccccccttt ccagttccga gagccaaggc ccatggatga actggtgacc ctggaggagg
     2041 ctgatggggg ctctgacatc ctgctggttg tccccaaggc aactgtgtta cagaaccagc
     2101 tcgatgagag ccagcaagaa cggaatgacc tgatgcagct gaagctacag ctggagggac
     2161 aggtgacaga gctgaggagc cgagtgcagg agctcgagag ggctctggca actgccaggc
     2221 aggagcacac ggagctgatg gaacagtaca aggggatttc ccggtcccat ggggagatca
     2281 cagaagagag ggacatcctg agccggcaac agggagacca tgtggcacgc atcctggagc
     2341 tagaggatga catccagacc atcagtgaga aagtgctgac gaaggaagtg gagctggaca
     2401 ggcttagaga cacagtgaag gccctgactc gggaacaaga gaagctcctt gggcaactga
     2461 aagaagtaca agcagacaag gagcaaagtg aggctgagct ccaagtggca caacaggaga
     2521 accatcactt aaatttggac ctgaaggagg cgaagagctg gcaagaggag cagagtgctc
     2581 aggctcagcg actgaaagac aaggtggccc agatgaagga caccctaggc caggcccagc
     2641 agcgggtggc cgagctggag cccttgaagg agcagcttcg aggggcccag gagcttgcag
     2701 cctcaagcca gcagaaagcc acccttcttg gggaggagtt ggccagcgca gcagcagcca
     2761 gggaccgcac catagccgaa ctacaccgca gccgcctgga agtggctgaa gttaacggca
     2821 ggctggctga gctcggtttg cacttgaagg aagaaaaatg ccaatggagc aaggagcggg
     2881 cagggctgct gcagagtgtg gaggcagaga aggacaagat cctgaagctg agtgcagaga
     2941 tacttcgatt ggagaaggca gttcaggagg agaggaccca aaaccaagtg ttcaagactg
     3001 agctggcccg ggagaaggat tctagcctgg tacagttgtc agaaagtaag cgggagctga
     3061 cagagctgcg gtcagccctg cgtgtgctcc agaaggaaaa ggagcagtta caggaggaga
     3121 aacaggaatt gctagagtac atgagaaagc tagaggcccg cctggagaag gtggcagatg
     3181 agaagtggaa tgaggatgcc accacagagg atgaggaggc cgctgtgggg ctgagctgcc
     3241 cggcagctct gacagactca gaggacgagt ccccagaaga catgaggctc ccaccctatg
     3301 gcctttgtga gcgtggagac ccaggctcct ctcctgctgg gcctcgagag gcttctcccc
     3361 ttgttgtcat cagccagccg gctcccattt ctcctcacct ctctgggcca gctgaggaca
     3421 gtagctctga ctcggaggct gaagatgaga agtcagtcct gatggcagct gtgcagagtg
     3481 ggggtgagga ggccaactta ctgcttcctg aactgggcag tgccttctat gacatggcca
     3541 gtggctttac agtgggtacc ctgtcagaaa ccagcactgg gggccctgcc acccccacat
     3601 ggaaggagtg tcctatctgt aaggagcgct ttcctgctga gagtgacaag gatgccctgg
     3661 aggaccacat ggatggacac ttctttttca gcacccagga ccccttcacc tttgagtgat
     3721 ctagaatgga gcagaagctg atctccgagg aggacctgtg aaagcttggc cgccatggcc
     3781 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3841 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3901 ttatcatgtc tggatcggga attaattcgg cgcagcacca tggcctgaaa taacctctga
     3961 aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca
     4021 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4081 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4141 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4201 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4261 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4321 tggaggccta ggcttttgca aaaagctgtt aacagcttgg cactggccgt cgttttacaa
     4381 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     4441 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     4501 agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4561 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     4621 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4681 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4741 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4801 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4861 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4921 accctatctc gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4981 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     5041 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     5101 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     5161 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     5221 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     5281 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5341 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5401 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5461 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5521 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5581 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5641 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5701 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5761 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5821 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5881 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5941 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6001 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6061 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6121 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6181 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6241 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6301 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6361 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6421 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6481 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6541 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6601 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6661 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6721 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6781 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6841 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6901 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6961 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7021 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7081 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     7141 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     7201 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     7261 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     7321 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     7381 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     7441 gaaacagcta tgacatgatt acgaattaa
//
