LOCUS       sequence-442326-        7130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Synchronize the trafficking of LAMP1-A17 from the ER..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7130)
  AUTHORS   Ecard J, Lian YL, Divoux S, Gouveia Z, Vigne E, Perez F, Boncompain 
            G
  TITLE     Lysosomal membrane proteins LAMP1 and LIMP2 are segregated in the 
            Golgi apparatus independently of their clathrin adaptor binding 
            motif.
  JOURNAL   Mol Biol Cell. 2024 Mar 1;35(3):ar42. doi: 10.1091/mbc.E23-06-0251. 
            Epub 2024 Jan 17.
  PUBMED    38231876
REFERENCE   2  (bases 1 to 7130)
  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: "Mol Biol 
            Cell."; date: "2024-03-1"; volume: "35(3):ar42. doi"; pages: " 
            10.1091/mbc.E23-06-0251. Epub 2024 Jan 17"
COMMENT     Sequence Label: Str-KDEL_SBP-mCherry-LAMP1-Y414A
FEATURES             Location/Qualifiers
     source          1..7130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          1646..1875
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1858..1877
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(1947..2521,2522..2524,2525..2533)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2114..2131)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2341..2360
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     sig_peptide     2547..2606
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             2607..2717
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="DEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     CDS             2727..3431
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2869..2887)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3137..3156
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4750..4871
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4787..4806)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4973..4989)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4973..4989)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4986..5008)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4997..5013
                     /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)."
     promoter        complement(join(5021..5027,5028..5045,5046..5051))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5066..5087
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5204..5221)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5375..5963)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5455..5474)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6134..6925,6926..6994))
                     /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     6757..6776
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6995..7099)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgatatctgc ggcctagcta gcgcttaagg cctgttaacc ggtatggatg
      961 tatgcgtccg tcttgccctg tggctcctct ggggactcct cctgcaccag ggccagagcc
     1021 tcagccatag tcacagtgag aaggcgacag gaaccagctc gggcgccggt ttaaacgacc
     1081 ctagcaaaga ctcaaaagct caggtgtccg ctgccgaggc tggcattact ggaacatggt
     1141 acaatcagct cgggagcacc tttattgtga ctgctggagc cgatggagcc ctcaccggaa
     1201 catacgaatc tgctgtggga aacgccgaat cacggtacgt cctcactggc cgatacgata
     1261 gtgcccctgc caccgacgga tctgggactg ccctgggatg gactgtcgct tggaaaaaca
     1321 actaccggaa tgctcattct gccacaacat ggagtggaca gtacgtggga ggcgctgagg
     1381 ctagaatcaa tacacagtgg ctgctcacat ctggcacaac cgaggcaaat gcttggaaat
     1441 ccaccctggt gggacatgac acattcacca aagtgaaacc ctccgccgct tcaatcgatg
     1501 ccgccaaaaa agccggagtc aacaacggca atcctctgga tgccgtccag cagctcggac
     1561 cgaaagatga actgtgagcg gccgcataga taactgatcc agtgtgctgg aattaattcg
     1621 ctgtctgcga gggccagctg ttggggtgag tactccctct caaaagcggg catgacttct
     1681 gcgctaagat tgtcagtttc caaaaacgag gaggatttga tattcacctg gcccgcggtg
     1741 atgcctttga gggtggccgc gtccatctgg tcagaaaaga caatcttttt gttgtcaagc
     1801 ttgaggtgtg gcaggcttga gatctggcca tacacttgag tgacaatgac atccactttg
     1861 cctttctctc cacaggtgtc cactcccagg tccaactgca ggtcgagcat gcatctaggg
     1921 cggccaattc cgcccctctc cccccccccc tctccctccc ccccccctaa cgttactggc
     1981 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     2041 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     2101 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     2161 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     2221 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     2281 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     2341 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     2401 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     2461 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     2521 gcttgccaca acccgggagg cgcgccatgt acaggatgca actcctgtct tgcattgcac
     2581 taagtcttgc acttgtcacg aattccgacg agaagaccac tggttggcga ggtggacacg
     2641 ttgttgaagg actggctggg gaacttgaac aacttcgtgc acgactggag catcacccac
     2701 aaggtcaacg tgaaccacct gcaggtgtga gcaagggcga ggaggataac atggccatca
     2761 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
     2821 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     2881 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
     2941 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     3001 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     3061 cccaggactc ctccctacag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     3121 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     3181 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     3241 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     3301 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     3361 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     3421 agctgtacaa gggccggcca gcaatgttta tggtgaaaaa tggcaacggg accgcgtgca
     3481 taatggccaa cttctctgct gccttctcag tgaactacga caccaagagt ggccctaaga
     3541 acatgacctt tgacctgcca tcagatgcca cagtggtgct caaccgcagc tcctgtggaa
     3601 aagagaacac ttctgacccc agtctcgtga ttgcttttgg aagaggacat acactcactc
     3661 tcaatttcac gagaaatgca acacgttaca gcgtccagct catgagtttt gtttataact
     3721 tgtcagacac acaccttttc cccaatgcga gctccaaaga aatcaagact gtggaatcta
     3781 taactgacat cagggcagat atagataaaa aatacagatg tgttagtggc acccaggtcc
     3841 acatgaacaa cgtgaccgta acgctccatg atgccaccat ccaggcgtac ctttccaaca
     3901 gcagcttcag caggggagag acacgctgtg aacaagacag gccttcccca accacagcgc
     3961 cccctgcgcc acccagcccc tcgccctcac ccgtgcccaa gagcccctct gtggacaagt
     4021 acaacgtgag cggcaccaac gggacctgcc tgctggccag catggggctg cagctgaacc
     4081 tcacctatga gaggaaggac aacacgacgg tgacaaggct tctcaacatc aaccccaaca
     4141 agacctcggc cagcgggagc tgcggcgccc acctggtgac tctggagctg cacagcgagg
     4201 gcaccaccgt cctgctcttc cagttcggga tgaatgcaag ttctagccgg tttttcctac
     4261 aaggaatcca gttgaataca attcttcctg acgccagaga ccctgccttt aaagctgcca
     4321 acggctccct gcgagcgctg caggccacag tcggcaattc ctacaagtgc aacgcggagg
     4381 agcacgtccg tgtcacgaag gcgttttcag tcaatatatt caaagtgtgg gtccaggctt
     4441 tcaaggtgga aggtggccag tttggctctg tggaggagtg tctgctggac gagaacagca
     4501 tgctgatccc catcgctgtg ggtggtgccc tggcggggct ggtcctcatc gtcctcatcg
     4561 cctacctcgt cggcaggaag aggagtcacg caggcgccca gactatctag ggcctcgagg
     4621 gccttaatta actctagata actgatcata atcagccata ccacatttgt tgaggtttta
     4681 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     4741 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4801 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4861 aatgtatctt aacgcgtcga gtgcattcta gttgtggttt gtccaaactc atcaatgtat
     4921 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     4981 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5041 taaggtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5101 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5161 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5221 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5281 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5341 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5401 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5461 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5521 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5581 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5641 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5701 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5761 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5821 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5881 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5941 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6001 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6061 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6121 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6181 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6241 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6301 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6361 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6421 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6481 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6541 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6601 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6661 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6721 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6781 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6841 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6901 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6961 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7021 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7081 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
