LOCUS       sequence-439313-        5961 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Localization: Lysosome Membrane, Excitation: 433, Emission: 475.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5961)
  AUTHORS   Markwardt ML, Kremers GJ, Kraft CA, Ray K, Cranfill PJ, Wilson KA, 
            Day RN, Wachter RM, Davidson MW, Rizzo MA
  TITLE     An improved cerulean fluorescent protein with enhanced brightness 
            and reduced reversible photoswitching.
  JOURNAL   PLoS One. 2011  . 6(3):e17896.
  PUBMED    21479270
REFERENCE   2  (bases 1 to 5961)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One. 
            2011  . 6(3):e17896."
COMMENT     Sequence Label: mCerulean3-Lysosomes-20
FEATURES             Location/Qualifiers
     source          1..5961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        38..341
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        342..545
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     496..516
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      597..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      601..610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(603..665,666..1823)
                     /codon_start=1
                     /product="rat lysosome-associated membrane glycoprotein 1"
                     /label=LAMP1
                     /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF
                     LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT
                     RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI
                     VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT
                     CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG
                     MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL
                     ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR
                     SHAGYQTI"
     regulatory      1878..1887
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1884..1886,1887..1889,1890..2603)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIK
                     ANFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1929..1950)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2190..2209)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2537..2558
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2726..2847
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2763..2782)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2817..2836
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2854..3309)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2991..3012)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3203..3222
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3336..3440
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3442..3799
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3442..3633
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3465..3485)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3650..3785
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3834..4628
                     /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             3834..4628
                     /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(3888..3907)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4498..4517
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4816..4835)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4860..4907
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5236..5824
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5725..5744
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
       61 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      121 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      181 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      241 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      301 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      361 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      421 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      481 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      541 gagctggttt agtgaaccgt cagatccgct agcgctaccg gactcagatc tcgagcgcca
      601 ccatggcggc cccgggcgcc cggcgtccgc tgctcctgtt gctgctggca ggccttgcac
      661 acagcgcccc agcactgttc gaggtgaaag acaacaacgg cacagcgtgt ataatggcca
      721 gcttctctgc ctcctttctg accacctatg aggctggaca tgtttctaag gtctcgaata
      781 tgaccctgcc agcctctgca gaagtcctga agaatagcag ctcttgtggt gaaaagaatg
      841 cttctgagcc caccctcgca atcacctttg gagaaggata tttactgaaa ctcaccttca
      901 caaaaaacac aacacgttac agtgtccagc acatgtattt cacatataac ctgtcagaca
      961 cacaattctt tcccaatgcc agctccaaag ggcccgacac tgtggattcc acaactgaca
     1021 tcaaggcaga catcaacaaa acataccgat gtgtcagcga catcagggtc tacatgaaga
     1081 atgtgaccat tgtgctctgg gacgctacta tccaggccta cctgccgagt agcaacttca
     1141 gcaaggaaga gacacgctgc ccacaggatc aaccttcccc aactactggg ccacccagcc
     1201 cctcaccacc acttgtgccc acaaacccca gtgtgtccaa gtacaatgtg actggtgaca
     1261 atggaacctg cctgctggcc tctatggcac tgcaactcaa catcacctac atgaagaagg
     1321 acaacacgac tgtgaccaga gcattcaaca tcaacccaag tgacaaatat agtgggactt
     1381 gcggtgccca gttggtgacc ctgaaggtgg ggaacaagag cagagtcctg gagctgcagt
     1441 ttgggatgaa tgccacttct agcctgtttt tcctgcaagg agttcagttg aacatgactc
     1501 ttcctgatgc catagagccc acgttcagca cctccaacta ttccctgaaa gctcttcagg
     1561 ccagtgtcgg caactcatac aagtgcaaca gtgaggagca catctttgtc agcaaggcgc
     1621 tcgccctcaa tgtcttcagc gtgcaagtcc aggctttcag ggtagaaagt gacaggtttg
     1681 ggtctgtgga agagtgtgta caggacggta acaacatgct gatccccatt gctgtgggcg
     1741 gggccctggc agggctggtc ctcatcgtcc tcatcgccta cctcatcggc aggaagagga
     1801 gtcacgcggg ctatcagacc atctcggaat tcggctccac cggctccacc ggctccaccg
     1861 gcgcggatcc accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     1921 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1981 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     2041 agctgcccgt gccctggccc accctcgtga ccaccctgag ctggggcgtg cagtgcttcg
     2101 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2161 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2221 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2281 aggacggcaa catcctgggg cacaagctgg agtacaacgc catccacggc aacgtctata
     2341 tcaccgccga caagcagaag aacggcatca aggccaactt cggcctcaac tgcaacatcg
     2401 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2461 ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca
     2521 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2581 gcatggacga gctgtacaag taaagcggcc gcgactctag atcataatca gccataccac
     2641 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     2701 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     2761 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2821 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
     2881 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     2941 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     3001 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     3061 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     3121 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     3181 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     3241 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     3301 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3361 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3421 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     3481 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3541 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3601 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3661 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3721 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3781 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     3841 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3901 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3961 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     4021 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4081 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4141 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4201 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4261 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4321 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     4381 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4441 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4501 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4561 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4621 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4681 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4741 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4801 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     4861 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     4921 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     4981 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     5041 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     5101 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5161 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5221 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5281 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5341 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5401 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5461 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5521 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5581 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5641 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5701 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5761 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5821 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5881 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
     5941 gttattaata gtaatcaatt a
//
