LOCUS       sequence-456534-        5960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Lysosome Membrane, Excitation: 485, Emission: 507.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5960)
  TITLE     Davidson Green fluorescent proteins
REFERENCE   2  (bases 1 to 5960)
  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: sfGFP-Lysosomes-20
FEATURES             Location/Qualifiers
     source          1..5960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     37..57
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      138..147
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      142..151
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(144..206,207..1364)
                     /codon_start=1
                     /product="rat lysosome-associated membrane glycoprotein 1"
                     /label=LAMP1
                     /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF
                     LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT
                     RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI
                     VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT
                     CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG
                     MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL
                     ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR
                     SHAGYQTI"
     regulatory      1419..1428
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1425..1427,1428..1430,1431..2144)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1470..1491)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2078..2099
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2266..2387
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2303..2322)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2357..2376
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2394..2849)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2531..2552)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2743..2762
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2876..2980
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2982..3339
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2982..3173
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3005..3025)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3190..3325
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3374..4168
                     /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             3374..4168
                     /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(3428..3447)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4038..4057
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4356..4375)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4400..4447
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4776..5364
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5265..5284
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5539..5842
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(5843..5960,1..86)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
       61 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      121 ggactcagat ctcgagcgcc accatggcgg ccccgggcgc ccggcgtccg ctgctcctgt
      181 tgctgctggc aggccttgca cacagcgccc cagcactgtt cgaggtgaaa gacaacaacg
      241 gcacagcgtg tataatggcc agcttctctg cctcctttct gaccacctat gaggctggac
      301 atgtttctaa ggtctcgaat atgaccctgc cagcctctgc agaagtcctg aagaatagca
      361 gctcttgtgg tgaaaagaat gcttctgagc ccaccctcgc aatcaccttt ggagaaggat
      421 atttactgaa actcaccttc acaaaaaaca caacacgtta cagtgtccag cacatgtatt
      481 tcacatataa cctgtcagac acacaattct ttcccaatgc cagctccaaa gggcccgaca
      541 ctgtggattc cacaactgac atcaaggcag acatcaacaa aacataccga tgtgtcagcg
      601 acatcagggt ctacatgaag aatgtgacca ttgtgctctg ggacgctact atccaggcct
      661 acctgccgag tagcaacttc agcaaggaag agacacgctg cccacaggat caaccttccc
      721 caactactgg gccacccagc ccctcaccac cacttgtgcc cacaaacccc agtgtgtcca
      781 agtacaatgt gactggtgac aatggaacct gcctgctggc ctctatggca ctgcaactca
      841 acatcaccta catgaagaag gacaacacga ctgtgaccag agcattcaac atcaacccaa
      901 gtgacaaata tagtgggact tgcggtgccc agttggtgac cctgaaggtg gggaacaaga
      961 gcagagtcct ggagctgcag tttgggatga atgccacttc tagcctgttt ttcctgcaag
     1021 gagttcagtt gaacatgact cttcctgatg ccatagagcc cacgttcagc acctccaact
     1081 attccctgaa agctcttcag gccagtgtcg gcaactcata caagtgcaac agtgaggagc
     1141 acatctttgt cagcaaggcg ctcgccctca atgtcttcag cgtgcaagtc caggctttca
     1201 gggtagaaag tgacaggttt gggtctgtgg aagagtgtgt acaggacggt aacaacatgc
     1261 tgatccccat tgctgtgggc ggggccctgg cagggctggt cctcatcgtc ctcatcgcct
     1321 acctcatcgg caggaagagg agtcacgcgg gctatcagac catctcggaa ttcggctcca
     1381 ccggctccac cggctccacc ggcgcggatc caccggtcgc caccatggtg agcaagggcg
     1441 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     1501 acaagttcag cgtgcgcggc gagggcgagg gcgatgccac caacggcaag ctgaccctga
     1561 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     1621 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcgccac gacttcttca
     1681 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cagcttcaag gacgacggca
     1741 cctacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     1801 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     1861 tcaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     1921 tcaagatccg ccacaacgtg gaggacggca gcgtgcagct cgccgaccac taccagcaga
     1981 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     2041 ccgtgctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     2101 ccgccgccgg gatcactcac ggcatggacg agctgtacaa gtaagcggcc gcgactctag
     2161 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     2221 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     2281 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2341 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     2401 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2461 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     2521 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     2581 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     2641 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     2701 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     2761 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     2821 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     2881 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2941 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     3001 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3061 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3121 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3181 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3241 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3301 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     3361 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3421 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3481 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3541 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     3601 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     3661 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3721 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3781 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3841 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3901 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     3961 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4021 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4081 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4141 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4201 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4261 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4321 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     4381 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     4441 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     4501 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     4561 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     4621 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4681 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4741 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4801 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4861 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4921 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4981 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5041 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5101 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5161 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5221 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5281 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5341 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5401 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5461 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
     5521 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     5581 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     5641 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     5701 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     5761 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     5821 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     5881 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     5941 ttggcaccaa aatcaacggg
//
