LOCUS       pCMV-mGold2t-Lysosomes- 5961 bp ds-DNA     circular     05-DEC-2024
DEFINITION  .
KEYWORDS    "accession:addgene_56618_241076"
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
COMMENT     Imported from database: AddGene Entry: mVenus-Lysosomes-M-20 URL:
            https://www.addgene.org/browse/sequence/241076/ GENERAL Depositing
            Lab: Michael Davidson Publication: Davidson Yellow fluorescent
            proteins (unpublished) Purpose: Localization: Lysosome Membrane,
            Excitation: 515, Emission: 528 BACKBONE Backbone size w/o insert
            (bp): 4750 Selectable markers: Neomycin (select with G418) Vector
            backbone: mVenus Vector type: Mammalian Expression CLONING
            INFORMATION 3′ sequencing primer: SV40pA-R 5′ sequencing primer:
            CMV-F Cloning method: Unknown GROWTH IN BACTERIA Bacterial
            Resistance(s): Kanamycin Copy number: High Copy Growth Strain(s):
            DH5alpha Growth Temperature: 37°C GENE/INSERT Alt name: NM_012857.2
            Gene/Insert name: LAMP1 M Mutation: R8 silent mutation; CGT
            Promoter: CMV Species: R. norvegicus (rat) Tag / Fusion Protein:
FEATURES             Location/Qualifiers
     primer_bind     102..122
                     /label="CMV-F"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     regulatory      203..212
                     /label="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     regulatory      207..216
                     /label="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     CDS             209..1429
                     /label="LAMP1"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    1463..1509
                     /label="5557_F"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     regulatory      1484..1489
                     /label="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    1490..1509
                     /label="5554_F"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    1490..1509
                     /label="5556_F"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     source          1490..2206
                     /label="source"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     CDS             1490..2206
                     /label="mGold2t"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     source          1490..2206
                     /label="source"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     source          1490..2206
                     /label="source"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     CDS             1490..2206
                     /label="Translation 1490-2206"
                     /translation="MASKGEELFTGVVPILVELDGDINGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTSLGYGLQCFARYPDNMKRHDFFKSAMPEGYVQERTIFFEDDGNYKTRAEVKFEGGTLVNRVELKGIDFKEDGNILGHKLEYNYNCHNVFITADRQQNGIKVNFKIRHNIEVGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQTKLSKDPNEKRDHMVLLEFVTAAGITLSMDELYK"
     CDS             1490..2206
                     /label="Translation 1490-2206"
                     /translation="MASKGEELFTGVVPILVELDGDINGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTSLGYGLQCFARYPDNMKRHDFFKSAMPEGYVQERTIFFEDDGNYKTRAEVKFEGGTLVNRVELKGIDFKEDGNILGHKLEYNYNCHNVFITADRQQNGIKVNFKIRHNIEVGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQTKLSKDPNEKRDHMVLLEFVTAAGITLSMDELYK"
     CDS             1793..1795
                     /label="Translation 1793-1795"
                     /translation="E"
     CDS             2009..2011
                     /label="Translation 2009-2011"
                     /translation="V"
     misc_feature    complement(2187..2206)
                     /label="3695_R"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    complement(2187..2206)
                     /label="3696_R"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    complement(2187..2230)
                     /label="3676_R"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     CDS             2207..2209
                     /label="mVenus"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     polyA_signal    2332..2453
                     /label="SV40 poly(A) signal"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer_bind     complement(2369..2388)
                     /label="SV40pA-R"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer_bind     2423..2442
                     /label="EBV-rev"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     rep_origin      complement(2460..2915)
                     /label="f1 ori"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer_bind     complement(2597..2618)
                     /label="F1ori-F"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     primer_bind     2809..2828
                     /label="F1ori-R"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     promoter        2942..3046
                     /label="AmpR promoter"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     promoter        3048..3405
                     /label="SV40 promoter"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer_bind     complement(3071..3091)
                     /label="pBABE 3'"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     rep_origin      3256..3391
                     /label="SV40 ori"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer_bind     3318..3337
                     /label="SV40pro-F"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     CDS             3440..4234
                     /label="NeoR/KanR"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer_bind     complement(3494..3513)
                     /label="Neo-R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer_bind     4104..4123
                     /label="Neo-F"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     primer_bind     complement(4422..4441)
                     /label="TK-pA-R"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     polyA_signal    4466..4513
                     /label="HSV TK poly(A) signal"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     rep_origin      4842..5430
                     /label="ori"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     primer_bind     5331..5350
                     /label="pBR322ori-F"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     source          5528..5527
                     /label="source"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     enhancer        5605..5908
                     /label="CMV enhancer"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     promoter        5909..151
                     /label="CMV promoter"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
ORIGIN
        1 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
       61 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      121 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg
      181 ctaccggact cagatctcga gcgccaccat ggcggccccg ggcgcccggc gtccgctgct
      241 cctgttgctg ctggcaggcc ttgcacacag cgccccagca ctgttcgagg tgaaagacaa
      301 caacggcaca gcgtgtataa tggccagctt ctctgcctcc tttctgacca cctatgaggc
      361 tggacatgtt tctaaggtct cgaatatgac cctgccagcc tctgcagaag tcctgaagaa
      421 tagcagctct tgtggtgaaa agaatgcttc tgagcccacc ctcgcaatca cctttggaga
      481 aggatattta ctgaaactca ccttcacaaa aaacacaaca cgttacagtg tccagcacat
      541 gtatttcaca tataacctgt cagacacaca attctttccc aatgccagct ccaaagggcc
      601 cgacactgtg gattccacaa ctgacatcaa ggcagacatc aacaaaacat accgatgtgt
      661 cagcgacatc agggtctaca tgaagaatgt gaccattgtg ctctgggacg ctactatcca
      721 ggcctacctg ccgagtagca acttcagcaa ggaagagaca cgctgcccac aggatcaacc
      781 ttccccaact actgggccac ccagcccctc accaccactt gtgcccacaa accccagtgt
      841 gtccaagtac aatgtgactg gtgacaatgg aacctgcctg ctggcctcta tggcactgca
      901 actcaacatc acctacatga agaaggacaa cacgactgtg accagagcat tcaacatcaa
      961 cccaagtgac aaatatagtg ggacttgcgg tgcccagttg gtgaccctga aggtggggaa
     1021 caagagcaga gtcctggagc tgcagtttgg gatgaatgcc acttctagcc tgtttttcct
     1081 gcaaggagtt cagttgaaca tgactcttcc tgatgccata gagcccacgt tcagcacctc
     1141 caactattcc ctgaaagctc ttcaggccag tgtcggcaac tcatacaagt gcaacagtga
     1201 ggagcacatc tttgtcagca aggcgctcgc cctcaatgtc ttcagcgtgc aagtccaggc
     1261 tttcagggta gaaagtgaca ggtttgggtc tgtggaagag tgtgtacagg acggtaacaa
     1321 catgctgatc cccattgctg tgggcggggc cctggcaggg ctggtcctca tcgtcctcat
     1381 cgcctacctc atcggcagga agaggagtca cgcgggctat cagaccatct cggaattcgg
     1441 ctccaccggc tccaccggct ccaccggcgc ggatccaccg gtcgccaccA TGGCGAGCAA
     1501 GGGCGAGGAG CTGTTCACCG GGGTGGTGCC CATCCTGGTC GAGCTGGACG GCGACATAAA
     1561 CGGCCACAAG TTCAGCGTGT CCGGCGAGGG CGAGGGCGAT GCCACCTACG GCAAGCTGAC
     1621 CCTGAAGTTC ATCTGCACCA CCGGCAAGCT GCCCGTGCCC TGGCCCACCC TCGTGACCAG
     1681 CCTGGGCTAC GGCCTGCAGT GCTTCGCCCG CTACCCCGAC AACATGAAGC GGCACGACTT
     1741 CTTCAAGTCC GCCATGCCCG AAGGCTACGT CCAGGAGCGC ACCATCTTCT TCGAGGACGA
     1801 CGGCAACTAC AAGACCCGCG CCGAGGTGAA GTTTGAGGGC GGCACCCTGG TGAACCGCGT
     1861 CGAGCTGAAG GGCATCGACT TCAAGGAGGA CGGCAACATC CTGGGGCACA AGCTGGAGTA
     1921 CAACTACAAC TGCCACAACG TCTTTATCAC CGCCGACAGG CAGCAGAACG GCATCAAGGT
     1981 CAACTTCAAG ATCCGCCACA ACATCGAGGT CGGCGGCGTG CAGCTCGCCG ACCACTACCA
     2041 GCAGAACACC CCCATCGGCG ACGGCCCCGT GCTGCTGCCC GACAACCACT ACCTGAGCTA
     2101 CCAGACCAAG CTGAGCAAAG ACCCCAACGA GAAGCGCGAT CACATGGTCC TGCTGGAGTT
     2161 CGTGACCGCC GCCGGGATCA CTCTCAGCAT GGACGAGCTG TACAAGtaaa gcggccgcga
     2221 ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2281 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2341 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2401 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     2461 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     2521 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2581 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2641 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     2701 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     2761 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2821 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     2881 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     2941 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3001 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     3061 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3121 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3181 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3241 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3301 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3361 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     3421 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3481 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3541 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3601 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3661 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3721 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3781 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3841 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     3901 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     3961 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4021 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4081 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4141 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4201 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4261 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4321 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4381 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     4441 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     4501 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     4561 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     4621 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     4681 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4741 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4801 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4861 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4921 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4981 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5041 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5101 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5161 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5221 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5281 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5341 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5401 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5461 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5521 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt
     5581 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     5641 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     5701 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     5761 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     5821 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     5881 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     5941 tggatagcgg tttgactcac g
//