LOCUS       sequence-212979-        5945 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 5945)
  TITLE     Davidson Orange Fluorescent Proteins
REFERENCE   2  (bases 1 to 5945)
  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: mKOet-Lysosomes-20
FEATURES             Location/Qualifiers
     source          1..5945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        52..355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        356..559
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     510..530
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      611..620
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      615..624
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(617..679,680..1837)
                     /codon_start=1
                     /product="rat lysosome-associated membrane glycoprotein 1"
                     /label=LAMP1
                     /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF
                     LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT
                     RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI
                     VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT
                     CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG
                     MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL
                     ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR
                     SHAGYQTI"
     regulatory      1892..1901
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1925..2575
                     /codon_start=1
                     /product="monomeric Kusabira-Orange fluorescent protein 
                     (Karasawa et al., 2004)"
                     /label=mKO
                     /translation="MAVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRVT
                     MAKGGPMPFAFDLVSHVFCYGHRPFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSAS
                     VSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMYL
                     KLEGGGNHKCQFKTTYKAAKKILKMPGSHYISHRLVRKTEGNITELVEDAVAHS"
     polyA_signal    2724..2845
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2761..2780)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2815..2834
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2852..3307)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2989..3010)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3201..3220
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3334..3438
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3440..3797
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3440..3631
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3463..3483)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3648..3783
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3832..4626
                     /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             3832..4626
                     /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(3886..3905)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4496..4515
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4814..4833)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4858..4905
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5234..5822
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5723..5742
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
       61 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      121 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      301 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      361 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      421 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      481 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      541 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggactca
      601 gatctcgagc gccaccatgg cggccccggg cgcccggcgt ccgctgctcc tgttgctgct
      661 ggcaggcctt gcacacagcg ccccagcact gttcgaggtg aaagacaaca acggcacagc
      721 gtgtataatg gccagcttct ctgcctcctt tctgaccacc tatgaggctg gacatgtttc
      781 taaggtctcg aatatgaccc tgccagcctc tgcagaagtc ctgaagaata gcagctcttg
      841 tggtgaaaag aatgcttctg agcccaccct cgcaatcacc tttggagaag gatatttact
      901 gaaactcacc ttcacaaaaa acacaacacg ttacagtgtc cagcacatgt atttcacata
      961 taacctgtca gacacacaat tctttcccaa tgccagctcc aaagggcccg acactgtgga
     1021 ttccacaact gacatcaagg cagacatcaa caaaacatac cgatgtgtca gcgacatcag
     1081 ggtctacatg aagaatgtga ccattgtgct ctgggacgct actatccagg cctacctgcc
     1141 gagtagcaac ttcagcaagg aagagacacg ctgcccacag gatcaacctt ccccaactac
     1201 tgggccaccc agcccctcac caccacttgt gcccacaaac cccagtgtgt ccaagtacaa
     1261 tgtgactggt gacaatggaa cctgcctgct ggcctctatg gcactgcaac tcaacatcac
     1321 ctacatgaag aaggacaaca cgactgtgac cagagcattc aacatcaacc caagtgacaa
     1381 atatagtggg acttgcggtg cccagttggt gaccctgaag gtggggaaca agagcagagt
     1441 cctggagctg cagtttggga tgaatgccac ttctagcctg tttttcctgc aaggagttca
     1501 gttgaacatg actcttcctg atgccataga gcccacgttc agcacctcca actattccct
     1561 gaaagctctt caggccagtg tcggcaactc atacaagtgc aacagtgagg agcacatctt
     1621 tgtcagcaag gcgctcgccc tcaatgtctt cagcgtgcaa gtccaggctt tcagggtaga
     1681 aagtgacagg tttgggtctg tggaagagtg tgtacaggac ggtaacaaca tgctgatccc
     1741 cattgctgtg ggcggggccc tggcagggct ggtcctcatc gtcctcatcg cctacctcat
     1801 cggcaggaag aggagtcacg cgggctatca gaccatctcg gaattcggct ccaccggctc
     1861 caccggctcc accggcgcgg atccaccggt cgccaccatg gtgagcaagg gcgaggagaa
     1921 taacatggcc gtgattaaac cagagatgaa gatgaggtac tacatggacg gctccgtcaa
     1981 tgggcatgag ttcacaattg aaggtgaagg cacaggcaga ccttacgagg gacatcaaga
     2041 gatgacacta cgcgtcacaa tggccaaggg cgggccaatg cctttcgcgt ttgacttagt
     2101 gtcacacgtg ttctgttacg gccacagacc ttttactaaa tatccagaag agataccaga
     2161 ctatttcaaa caagcatttc ctgaaggcct gtcatgggaa aggtcgttgg agttcgaaga
     2221 tggtgggtcc gcttcagtca gtgcgcatat aagccttaga ggaaacacct tctaccacaa
     2281 atccaaattt actggggtta actttcctgc cgatggtcct atcatgcaaa accaaagtgt
     2341 tgattgggag ccatcaaccg agaaaattac tgccagcgac ggagttctga agggtgatgt
     2401 tacgatgtac ctaaaacttg aaggaggcgg caatcacaaa tgccaattca agactactta
     2461 caaggcggca aaaaagattc ttaaaatgcc aggaagccat tacatcagcc atcgcctcgt
     2521 caggaaaacc gaaggcaaca ttactgagct ggtagaagat gcagtagctc attccggcgg
     2581 catggacgag ctgtacaagt aagcggccgc gactctagat cataatcagc cataccacat
     2641 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     2701 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     2761 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     2821 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     2881 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     2941 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     3001 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     3061 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     3121 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     3181 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     3241 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     3301 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3361 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3421 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     3481 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3541 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3601 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3661 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3721 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3781 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     3841 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3901 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3961 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     4021 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4081 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4141 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4201 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4261 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4321 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     4381 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4441 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4501 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4561 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4621 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4681 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4741 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     4801 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     4861 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     4921 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     4981 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     5041 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     5101 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5161 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5221 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5281 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5341 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5401 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5461 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5521 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5581 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5641 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5701 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5761 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5821 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5881 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt
     5941 tatta
//
