LOCUS       sequence-251156-        5897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Lysosome Membrane, Excitation: 548, Emission: 561.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5897)
  TITLE     Davidson Orange Fluorescent Proteins
REFERENCE   2  (bases 1 to 5897)
  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"
COMMENT     Sequence Label: mKO-Lysosomes-20
FEATURES             Location/Qualifiers
     source          1..5897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        106..409
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        410..613
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     564..584
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      665..674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      669..678
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(671..733,734..1891)
                     /codon_start=1
                     /product="rat lysosome-associated membrane glycoprotein 1"
                     /label=LAMP1
                     /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF
                     LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT
                     RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI
                     VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT
                     CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG
                     MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL
                     ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR
                     SHAGYQTI"
     regulatory      1946..1955
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1952..2608
                     /codon_start=1
                     /product="monomeric Kusabira-Orange fluorescent protein 
                     (Karasawa et al., 2004)"
                     /label=mKO
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAKGGPMPFAFDLVSHVFCYGHRPFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKKILKMPGSHYISHRLVRKTEGNITELVEDAVAHS"
     polyA_signal    2730..2851
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2767..2786)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2821..2840
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2858..3313)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2995..3016)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3207..3226
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3340..3444
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3446..3803
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3446..3637
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3469..3489)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3654..3789
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3838..4632
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3838..4632
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3892..3911)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4502..4521
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4820..4839)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4864..4911
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5240..5828
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5729..5748
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
       61 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
      121 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      181 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      241 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      301 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      361 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
      421 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
      481 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      541 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      601 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga ctcagatctc
      661 gagcgccacc atggcggccc cgggcgcccg gcggccgctg ctcctgttgc tgctggcagg
      721 ccttgcacac agcgccccag cactgttcga ggtgaaagac aacaacggca cagcgtgtat
      781 aatggccagc ttctctgcct cctttctgac cacctatgag gctggacatg tttctaaggt
      841 ctcgaatatg accctgccag cctctgcaga agtcctgaag aatagcagct cttgtggtga
      901 aaagaatgct tctgagccca ccctcgcaat cacctttgga gaaggatatt tactgaaact
      961 caccttcaca aaaaacacaa cacgttacag tgtccagcac atgtatttca catataacct
     1021 gtcagacaca caattctttc ccaatgccag ctccaaaggg cccgacactg tggattccac
     1081 aactgacatc aaggcagaca tcaacaaaac ataccgatgt gtcagcgaca tcagggtcta
     1141 catgaagaat gtgaccattg tgctctggga cgctactatc caggcctacc tgccgagtag
     1201 caacttcagc aaggaagaga cacgctgccc acaggatcaa ccttccccaa ctactgggcc
     1261 acccagcccc tcaccaccac ttgtgcccac aaaccccagt gtgtccaagt acaatgtgac
     1321 tggtgacaat ggaacctgcc tgctggcctc tatggcactg caactcaaca tcacctacat
     1381 gaagaaggac aacacgactg tgaccagagc attcaacatc aacccaagtg acaaatatag
     1441 tgggacttgc ggtgcccagt tggtgaccct gaaggtgggg aacaagagca gagtcctgga
     1501 gctgcagttt gggatgaatg ccacttctag cctgtttttc ctgcaaggag ttcagttgaa
     1561 catgactctt cctgatgcca tagagcccac gttcagcacc tccaactatt ccctgaaagc
     1621 tcttcaggcc agtgtcggca actcatacaa gtgcaacagt gaggagcaca tctttgtcag
     1681 caaggcgctc gccctcaatg tcttcagcgt gcaagtccag gctttcaggg tagaaagtga
     1741 caggtttggg tctgtggaag agtgtgtaca ggacggtaac aacatgctga tccccattgc
     1801 tgtgggcggg gccctggcag ggctggtcct catcgtcctc atcgcctacc tcatcggcag
     1861 gaagaggagt cacgcgggct atcagaccat ctcggaattc ggctccaccg gctccaccgg
     1921 ctccaccggc gcggatccac cggtcgccac catggtgagt gtgattaaac cagagatgaa
     1981 gatgaggtac tacatggacg gctccgtcaa tgggcatgag ttcacaattg aaggtgaagg
     2041 cacaggcaga ccttacgagg gacatcaaga gatgacacta cgcgtcacaa tggccaaggg
     2101 cgggccaatg cctttcgcgt ttgacttagt gtcacacgtg ttctgttacg gccacagacc
     2161 ttttactaaa tatccagaag agataccaga ctatttcaaa caagcatttc ctgaaggcct
     2221 gtcatgggaa aggtcgttgg agttcgaaga tggtgggtcc gcttcagtca gtgcgcatat
     2281 aagccttaga ggaaacacct tctaccacaa atccaaattt actggggtta actttcctgc
     2341 cgatggtcct atcatgcaaa accaaagtgt tgattgggag ccatcaaccg agaaaattac
     2401 tgccagcgac ggagttctga agggtgatgt tacgatgtac ctaaaacttg aaggaggcgg
     2461 caatcacaaa tgccaattca agactactta caaggcggca aaaaagattc ttaaaatgcc
     2521 aggaagccat tacatcagcc atcgcctcgt caggaaaacc gaaggcaaca ttactgagct
     2581 ggtagaagat gcagtagctc attcctaagc ggccgcgact ctagatcata atcagccata
     2641 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     2701 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     2761 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2821 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg
     2881 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     2941 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     3001 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     3061 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     3121 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     3181 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     3241 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     3301 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3361 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3421 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     3481 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3541 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3601 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3661 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3721 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3781 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     3841 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3901 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3961 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     4021 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4081 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4141 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4201 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4261 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4321 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     4381 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4441 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4501 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4561 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4621 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4681 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4741 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4801 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     4861 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     4921 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     4981 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     5041 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     5101 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5161 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5221 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5281 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5341 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5401 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5461 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5521 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5581 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5641 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5701 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5761 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5821 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5881 ctcacatgtt ctttcct
//
