LOCUS       sequence-388049-        6633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of LAMP1-miniSOG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6633)
  TITLE     Hensel lab plasmids
REFERENCE   2  (bases 1 to 6633)
  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: p3806
FEATURES             Location/Qualifiers
     source          1..6633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     525..545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(641..724,725..1891)
                     /codon_start=1
                     /product="human lysosome-associated membrane glycoprotein 
                     1"
                     /label=LAMP1
                     /translation="MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMAN
                     FSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNF
                     TRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHM
                     NNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKY
                     NVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSE
                     GTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNA
                     EEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLIV
                     LIAYLVGRKRSHAGYQTI"
     regulatory      1907..1916
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1913..2230
                     /codon_start=1
                     /product="mini singlet oxygen generator, a fluorescent 
                     flavoprotein suitable for correlated light and electron 
                     microscopy (Shu et al., 2011)"
                     /label=miniSOG
                     /translation="MEKSFVITDPRLPDNPIIFASDGFLELTEYSREEILGRNGRFLQG
                     PETDQATVQKIRDAIRDQREITVQLINYTKSGKKFWNLLHLQPMRDQKGELQYFIGVQL
                     DG"
     CDS             2258..2575
                     /codon_start=1
                     /product="mini singlet oxygen generator, a fluorescent 
                     flavoprotein suitable for correlated light and electron 
                     microscopy (Shu et al., 2011)"
                     /label=miniSOG
                     /translation="MEKSFVITDPRLPDNPIIFASDGFLELTEYSREEILGRNGRFLQG
                     PETDQATVQKIRDAIRDQREITVQLINYTKSGKKFWNLLHLQPMRDQKGELQYFIGVQL
                     DG"
     regulatory      2588..2597
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2594..3304
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2739..2757)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3007..3026
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3427..3548
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3464..3483)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3518..3537
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3555..4010)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3692..3713)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3904..3923
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4037..4141
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4143..4500
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4143..4334
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4166..4186)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4351..4486
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4535..5329
                     /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             4535..5329
                     /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(4589..4608)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5199..5218
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5517..5536)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5561..5608
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5937..6525
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6426..6445
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgctaccgg actcagatct cgagctcaag cttcgaattc atggcggccc ccggcagcgc
      661 ccggcgaccc ctgctgctgc tactgctgtt gctgctgctc ggcctcatgc attgtgcgtc
      721 agcagcaatg tttatggtga aaaatggcaa cgggaccgcg tgcataatgg ccaacttctc
      781 tgctgccttc tcagtgaact acgacaccaa gagtggccct aagaacatga cctttgacct
      841 gccatcagat gccacagtgg tgctcaaccg cagctcctgt ggaaaagaga acacttctga
      901 ccccagtctc gtgattgctt ttggaagagg acatacactc actctcaatt tcacgagaaa
      961 tgcaacacgt tacagcgtcc agctcatgag ttttgtttat aacttgtcag acacacacct
     1021 tttccccaat gcgagctcca aagaaatcaa gactgtggaa tctataactg acatcagggc
     1081 agatatagat aaaaaataca gatgtgttag tggcacccag gtccacatga acaacgtgac
     1141 cgtaacgctc catgatgcca ccatccaggc gtacctttcc aacagcagct tcagcagggg
     1201 agagacacgc tgtgaacaag acaggccttc cccaaccaca gcgccccctg cgccacccag
     1261 cccctcgccc tcacccgtgc ccaagagccc ctctgtggac aagtacaacg tgagcggcac
     1321 caacgggacc tgcctgctgg ccagcatggg gctgcagctg aacctcacct atgagaggaa
     1381 ggacaacacg acggtgacaa ggcttctcaa catcaacccc aacaagacct cggccagcgg
     1441 gagctgcggc gcccacctgg tgactctgga gctgcacagc gagggcacca ccgtcctgct
     1501 cttccagttc gggatgaatg caagttctag ccggtttttc ctacaaggaa tccagttgaa
     1561 tacaattctt cctgacgcca gagaccctgc ctttaaagct gccaacggct ccctgcgagc
     1621 gctgcaggcc acagtcggca attcctacaa gtgcaacgcg gaggagcacg tccgtgtcac
     1681 gaaggcgttt tcagtcaata tattcaaagt gtgggtccag gctttcaagg tggaaggtgg
     1741 ccagtttggc tctgtggagg agtgtctgct ggacgagaac agcatgctga tccccatcgc
     1801 tgtgggtggt gccctggcgg ggctggtcct catcgtcctc atcgcctacc tcgtcggcag
     1861 gaagaggagt cacgcaggct accagactat ccgggatcca ccggtcgcca ccatggagaa
     1921 gagcttcgtg atcaccgacc cccgcctgcc cgacaacccc atcatcttcg ccagcgacgg
     1981 cttcctggag ctgaccgagt acagcagaga ggagatcctg ggccgcaacg gccgcttcct
     2041 gcagggcccc gagaccgacc aggccaccgt gcagaagatc cgcgacgcca tccgcgacca
     2101 gcgcgagatc accgtgcagc tgatcaacta caccaagagc ggcaagaagt tctggaacct
     2161 gctgcacctg cagcccatgc gcgaccagaa gggcgagctg cagtacttca tcggcgtgca
     2221 gctggacggc tctggtggcg ggagtggtgg cgggtccatg gagaaaagtt tcgtgataac
     2281 tgatccacgg ctgccagaca atcccatcat cttcgcatcc gatggcttcc tggagctgac
     2341 cgagtattcc agagaggaga tcctgggccg caatggccgc tttctgcagg gaccagagac
     2401 agaccaggcc acagtgcaga agattcgcga tgccattaga gatcagcgcg agattaccgt
     2461 gcagctgata aactacacaa aaagcgggaa gaaattctgg aacctcctgc acctccagcc
     2521 catgagggac cagaagggtg agctccagta tttcatcgga gtgcagctgg atggagatcc
     2581 accggtcgcc accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
     2641 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     2701 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     2761 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     2821 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     2881 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     2941 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     3001 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     3061 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     3121 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     3181 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     3241 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     3301 gtaaagcggc cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt
     3361 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3421 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3481 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3541 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3601 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3661 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3721 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     3781 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     3841 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3901 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     3961 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     4021 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4081 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4141 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4201 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4261 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4321 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4381 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4441 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4501 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4561 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4621 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4681 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     4741 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4801 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4861 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4921 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4981 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5041 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5101 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5161 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5221 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5281 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5341 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5401 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5461 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     5521 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     5581 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5641 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     5701 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     5761 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     5821 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5881 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5941 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6001 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6061 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6121 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6181 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6241 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6301 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6361 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6421 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6481 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6541 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6601 tcccctgatt ctgtggataa ccgtattacc gcc
//
