LOCUS       sequence-331260-        5987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express rat LAMP1-mRFP in zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5987)
  AUTHORS   Morishita H, Kanda Y, Kaizuka T, Chino H, Nakao K, Miki Y, Taketomi 
            Y, Guan JL, Murakami M, Aiba A, Mizushima N
  TITLE     Autophagy Is Required for Maturation of Surfactant-Containing 
            Lamellar Bodies in the Lung and Swim Bladder.
  JOURNAL   Cell Rep. 2020 Dec 8;33(10):108477. doi: 
            10.1016/j.celrep.2020.108477.
  PUBMED    33296658
REFERENCE   2  (bases 1 to 5987)
  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: "Cell Rep.";
            date: "2020-12-8"; volume: "33(10):108477. doi"; pages: " 
            10.1016/j.celrep.2020.108477"
COMMENT     Sequence Label: pCS2-LAMP1-mRFP
FEATURES             Location/Qualifiers
     source          1..5987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      90..99
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(92..154,155..1312)
                     /codon_start=1
                     /product="rat lysosome-associated membrane glycoprotein 1"
                     /label=LAMP1
                     /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF
                     LTTYDAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT
                     RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI
                     VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT
                     CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG
                     MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL
                     ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR
                     SHAGYQTI"
     CDS             1325..2002
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1325..2002
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1455..1473)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1723..1742
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1917..1940
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2039..2173
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2063..2082)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2117..2136
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2286..2306)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2286..2304)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2325..2341)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2325..2341)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2338..2360)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2349..2365
                     /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(2373..2379,2380..2397,2398..2403))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2418..2439
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2556..2573)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2727..3315)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2807..2826)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3486..4277,4278..4346))
                     /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     4109..4128
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4347..4451)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4477..4932)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4614..4635)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4826..4845
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5003..5987
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atctcgagac catggcggcc ccgggcgccc ggcggccgct
      121 gctcctgttg ctgctggcag gccttgcaca cagcgcccca gcactgttcg aggtgaaaga
      181 caacaacggc acagcgtgta taatggccag cttctctgcc tcctttctga ccacctatga
      241 tgctggacat gtttctaagg tctcgaatat gaccctgcca gcctctgcag aagtcctgaa
      301 gaatagcagc tcttgtggtg aaaagaatgc ttctgagccc accctcgcaa tcacctttgg
      361 agaaggatat ttactgaaac tcaccttcac aaaaaacaca acacgttaca gtgtccagca
      421 catgtatttc acatataacc tgtcagacac acaattcttt cccaatgcca gctccaaagg
      481 gcccgacact gtggattcca caactgacat caaggcagac atcaacaaaa cataccgatg
      541 tgtcagcgac atcagggtct acatgaagaa tgtgaccatt gtgctctggg acgctactat
      601 ccaggcctac ctgccgagta gcaacttcag caaggaagag acacgctgcc cacaggatca
      661 accttcccca actactgggc cacccagccc ctcaccacca cttgtgccca caaaccccag
      721 tgtgtccaag tacaatgtga ctggtgacaa tggaacctgc ctgctggcct ctatggcact
      781 gcaactcaac atcacctaca tgaagaagga caacacgact gtgaccagag cattcaacat
      841 caacccaagt gacaaatata gtgggacttg cggtgcccag ttggtgaccc tgaaggtggg
      901 gaacaagagc agagtcctgg agctgcagtt tgggatgaat gccacttcta gcctgttttt
      961 cctgcaagga gttcagttga acatgactct tcctgatgcc atagagccca cgttcagcac
     1021 ctccaactat tccctgaaag ctcttcaggc cagtgtcggc aactcataca agtgcaacag
     1081 tgaggagcac atctttgtca gcaaggcgct cgccctcaat gtcttcagcg tgcaagtcca
     1141 ggctttcagg gtagaaagtg acaggtttgg gtctgtggaa gagtgtgtac aggacggtaa
     1201 caacatgctg atccccattg ctgtgggcgg ggccctggca gggctggtcc tcatcgtcct
     1261 catcgcctac ctcatcggca ggaagaggag tcacgcgggc tatcagacca tcatcgatgg
     1321 taccatggcc tcctccgagg acgtcatcaa ggagttcatg cgcttcaagg tgcgcatgga
     1381 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
     1441 gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga
     1501 catcctgtcc cctcagttcc agtacggctc caaggcctac gtgaagcacc ccgccgacat
     1561 ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     1621 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat
     1681 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa
     1741 gaccatgggc tgggaggcct ccaccgagcg gatgtacccc gaggacggcg ccctgaaggg
     1801 cgagatcaag atgaggctga agctgaagga cggcggccac tacgacgccg aggtcaagac
     1861 cacctacatg gccaagaagc ccgtgcagct gcccggcgcc tacaagaccg acatcaagct
     1921 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgagc gcgccgaggg
     1981 ccgccactcc accggcgcct agctcgacct ctagaactat agtgagtcgt attacgtaga
     2041 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2101 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2161 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     2221 gtgggaggtt ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt
     2281 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     2341 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccgggag cataaagtgt
     2401 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     2461 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     2521 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2581 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2641 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2701 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2761 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2821 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2881 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2941 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3001 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3061 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3121 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3181 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3241 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3301 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3361 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3421 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3481 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3541 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3601 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3661 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3721 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3781 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3841 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3901 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3961 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4021 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4081 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4141 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4201 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4261 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4321 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4381 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4441 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     4501 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4561 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4621 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4681 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4741 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4801 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4861 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4921 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     4981 gggcctcttc gctattacgc cagtcgaccg ccaattcaat atggcgtata tggactcatg
     5041 ccaattcaat atggtggatc tggacctgtg ccaattcaat atggcgtata tggactcgtg
     5101 ccaattcaat atggtggatc tggaccccag ccaattcaat atggcggact tggcaccatg
     5161 ccaattcaat atggcggacc tggcactgtg ccaactgggg aggggtctac ttggcacggt
     5221 gccaagtttg aggaggggtc ttggccctgt gccaagtccg ccatattgaa ttggcatggt
     5281 gccaataatg gcggccatat tggctatatg ccaggatcaa tatataggca atatccaata
     5341 tggccctatg ccaatatggc tattggccag gttcaatact atgtattggc cctatgccat
     5401 atagtattcc atatatgggt tttcctattg acgtagatag cccctcccaa tgggcggtcc
     5461 catataccat atatggggct tcctaatacc gcccatagcc actcccccat tgacgtcaat
     5521 ggtctctata tatggtcttt cctattgacg tcatatgggc ggtcctattg acgtatatgg
     5581 cgcctccccc attgacgtca attacggtaa atggcccgcc tggctcaatg cccattgacg
     5641 tcaataggac cacccaccat tgacgtcaat gggatggctc attgcccatt catatccgtt
     5701 ctcacgcccc ctattgacgt caatgacggt aaatggccca cttggcagta catcaatatc
     5761 tattaatagt aacttggcaa gtacattact attggaagta cgccagggta cattggcagt
     5821 actcccattg acgtcaatgg cggtaaatgg cccgcgatgg ctgccaagta catccccatt
     5881 gacgtcaatg gggaggggca atgacgcaaa tgggcgttcc attgacgtaa atgggcggta
     5941 ggcgtgccta atgggaggtc tatataagca atgctcgttt agggaac
//
