LOCUS       sequence-331253-        6086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express rat LAMP1-EGFP in zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6086)
  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 6086)
  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-EGFP
FEATURES             Location/Qualifiers
     source          1..6086
                     /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"
     misc_feature    83..135
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      137..146
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(139..201,202..1359)
                     /codon_start=1
                     /product="rat lysosome-associated membrane glycoprotein 1"
                     /label=LAMP1
                     /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF
                     LTTYDAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT
                     RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI
                     VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT
                     CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG
                     MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL
                     ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR
                     SHAGYQTI"
     regulatory      1375..1384
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1381..1383,1384..1386,1387..2100)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1381..1383,1384..1386,1387..2097)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1426..1447)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1687..1706)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2034..2055
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2138..2272
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2162..2181)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2216..2235
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2385..2405)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2385..2403)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2424..2440)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2424..2440)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2437..2459)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2448..2464
                     /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(2472..2478,2479..2496,2497..2502))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2517..2538
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2655..2672)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2826..3414)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2906..2925)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3585..4376,4377..4445))
                     /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     4208..4227
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4446..4550)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4576..5031)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4713..4734)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4925..4944
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5102..6086
                     /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 atctcgagct caagcttcga attctgcagt cgacggtacc
      121 gcgggcccgg gatccaccat ggcggccccg ggcgcccggc ggccgctgct cctgttgctg
      181 ctggcaggcc ttgcacacag cgccccagca ctgttcgagg tgaaagacaa caacggcaca
      241 gcgtgtataa tggccagctt ctctgcctcc tttctgacca cctatgatgc tggacatgtt
      301 tctaaggtct cgaatatgac cctgccagcc tctgcagaag tcctgaagaa tagcagctct
      361 tgtggtgaaa agaatgcttc tgagcccacc ctcgcaatca cctttggaga aggatattta
      421 ctgaaactca ccttcacaaa aaacacaaca cgttacagtg tccagcacat gtatttcaca
      481 tataacctgt cagacacaca attctttccc aatgccagct ccaaagggcc cgacactgtg
      541 gattccacaa ctgacatcaa ggcagacatc aacaaaacat accgatgtgt cagcgacatc
      601 agggtctaca tgaagaatgt gaccattgtg ctctgggacg ctactatcca ggcctacctg
      661 ccgagtagca acttcagcaa ggaagagaca cgctgcccac aggatcaacc ttccccaact
      721 actgggccac ccagcccctc accaccactt gtgcccacaa accccagtgt gtccaagtac
      781 aatgtgactg gtgacaatgg aacctgcctg ctggcctcta tggcactgca actcaacatc
      841 acctacatga agaaggacaa cacgactgtg accagagcat tcaacatcaa cccaagtgac
      901 aaatatagtg ggacttgcgg tgcccagttg gtgaccctga aggtggggaa caagagcaga
      961 gtcctggagc tgcagtttgg gatgaatgcc acttctagcc tgtttttcct gcaaggagtt
     1021 cagttgaaca tgactcttcc tgatgccata gagcccacgt tcagcacctc caactattcc
     1081 ctgaaagctc ttcaggccag tgtcggcaac tcatacaagt gcaacagtga ggagcacatc
     1141 tttgtcagca aggcgctcgc cctcaatgtc ttcagcgtgc aagtccaggc tttcagggta
     1201 gaaagtgaca ggtttgggtc tgtggaagag tgtgtacagg acggtaacaa catgctgatc
     1261 cccattgctg tgggcggggc cctggcaggg ctggtcctca tcgtcctcat cgcctacctc
     1321 atcggcagga agaggagtca cgcgggctat cagaccatcg gggatccacc ggtcgccacc
     1381 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1441 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1501 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1561 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     1621 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     1681 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1741 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1801 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     1861 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     1921 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1981 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     2041 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     2101 ctcgagcctc tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat
     2161 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2221 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     2281 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc
     2341 ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta
     2401 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     2461 acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga
     2521 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2581 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2641 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2701 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2761 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2821 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2881 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2941 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3001 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3061 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3121 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3181 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3241 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3301 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3361 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3421 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3481 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3541 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3601 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3661 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3721 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3781 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3841 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3901 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3961 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4021 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4081 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4141 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4201 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4261 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4321 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4381 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4441 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4501 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4561 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     4621 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     4681 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4741 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4801 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4861 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4921 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     4981 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
     5041 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     5101 agtcgaccgc caattcaata tggcgtatat ggactcatgc caattcaata tggtggatct
     5161 ggacctgtgc caattcaata tggcgtatat ggactcgtgc caattcaata tggtggatct
     5221 ggaccccagc caattcaata tggcggactt ggcaccatgc caattcaata tggcggacct
     5281 ggcactgtgc caactgggga ggggtctact tggcacggtg ccaagtttga ggaggggtct
     5341 tggccctgtg ccaagtccgc catattgaat tggcatggtg ccaataatgg cggccatatt
     5401 ggctatatgc caggatcaat atataggcaa tatccaatat ggccctatgc caatatggct
     5461 attggccagg ttcaatacta tgtattggcc ctatgccata tagtattcca tatatgggtt
     5521 ttcctattga cgtagatagc ccctcccaat gggcggtccc atataccata tatggggctt
     5581 cctaataccg cccatagcca ctcccccatt gacgtcaatg gtctctatat atggtctttc
     5641 ctattgacgt catatgggcg gtcctattga cgtatatggc gcctccccca ttgacgtcaa
     5701 ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt caataggacc acccaccatt
     5761 gacgtcaatg ggatggctca ttgcccattc atatccgttc tcacgccccc tattgacgtc
     5821 aatgacggta aatggcccac ttggcagtac atcaatatct attaatagta acttggcaag
     5881 tacattacta ttggaagtac gccagggtac attggcagta ctcccattga cgtcaatggc
     5941 ggtaaatggc ccgcgatggc tgccaagtac atccccattg acgtcaatgg ggaggggcaa
     6001 tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa tgggaggtct
     6061 atataagcaa tgctcgttta gggaac
//
