LOCUS       sequence-355803-        7996 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of LAMP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7996)
  AUTHORS   Nabb AT, Bentley M
  TITLE     NgCAM and VAMP2 Reveal that Direct Delivery and Dendritic 
            Degradation Maintain Axonal Polarity.
  JOURNAL   Mol Biol Cell. 2021 Nov 3:mbcE21080425. doi: 
            10.1091/mbc.E21-08-0425.
  PUBMED    34731031
REFERENCE   2  (bases 1 to 7996)
  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"; journalName: "Mol Biol 
            Cell. 2021 Nov 3:mbcE21080425. doi: 10.1091/mbc.E21-08-0425."
COMMENT     Sequence Label: pBa.Lamp1-GFP
FEATURES             Location/Qualifiers
     source          1..7996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..276
                     /label=chicken beta-actin promoter
     primer_bind     1219..1241
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1307..1316
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1309..1392,1393..2559)
                     /codon_start=1
                     /product="human lysosome-associated membrane glycoprotein 
                     1"
                     /label=LAMP1
                     /translation="MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMAN
                     FSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNF
                     TRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHM
                     NNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKY
                     NVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSE
                     GTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNA
                     EEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSTLIPIAVGGALAGLVLIV
                     LIAYLVGRKRSHAGYQTI"
     CDS             join(2566..2568,2569..2571,2572..3282)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2566..2568,2569..2571,2572..3282)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2611..2632)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2872..2891)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3219..3240
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3415..3549
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3452..3471)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3506..3525
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3580..3596)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3580..3596)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3593..3615)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3604..3620
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3628..3634,3635..3652,3653..3658))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3673..3694
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3811..3828)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3982..4570)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4062..4081)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4741..5532,5533..5601))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5364..5383
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5602..5706)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5774..5792
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5830..5852)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5952..5971
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6165..6187
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6179..6196
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6180..6196
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
       61 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg
      121 cgcgcgccag gcggggcggg gcggggcgrg gggcggggcg gggcgaggcg gagaggtgcg
      181 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
      241 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
      301 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
      361 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
      421 taatgacggc tcgtttcttt tctgtggctg cgtgaaagcc ttaaagggct ccgggagggc
      481 cctttgtgcg ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc
      541 cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg
      601 tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg
      661 gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg
      721 tgtgggcgcg gcggtcgggc tgtaaccccc ccctgcaccc ccctccccga gttgctgagc
      781 acggcccggc ttcgggtgcg gggctccgtg cggggcgtgg cgcggggctc gccgtgccgg
      841 gcggggggtg gcggcaggkg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg
      901 gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc
      961 gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa
     1021 tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga
     1081 agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc
     1141 gccgtcccct tctccatctc cagcctcggg gctgtccgca gggggacggc tgccttcggg
     1201 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggggtt tatatcttcc
     1261 cttctctgtt cctccgcagc ccccaagctt ggcgcgccta ggcctaccat ggcggccccc
     1321 ggcagcgccc ggcgacccct gctgctgcta ctgctgttgc tgctgctcgg cctcatgcat
     1381 tgtgcgtcag cagcaatgtt tatggtgaaa aatggcaacg ggaccgcgtg cataatggcc
     1441 aacttctctg ctgccttctc agtgaactac gacaccaaga gtggccctaa gaacatgacc
     1501 tttgacctgc catcagatgc cacagtggtg ctcaaccgca gctcctgtgg aaaagagaac
     1561 acttctgacc ccagtctcgt gattgctttt ggaagaggac atacactcac tctcaatttc
     1621 acgagaaatg caacacgtta cagcgtccag ctcatgagtt ttgtttataa cttgtcagac
     1681 acacaccttt tccccaatgc gagctccaaa gaaatcaaga ctgtggaatc tataactgac
     1741 atcagggcag atatagataa aaaatacaga tgtgttagtg gcacccaggt ccacatgaac
     1801 aacgtgaccg taacgctcca tgatgccacc atccaggcgt acctttccaa cagcagcttc
     1861 agccggggag agacacgctg tgaacaagac aggccttccc caaccacagc gccccctgcg
     1921 ccacccagcc cctcgccctc acccgtgccc aagagcccct ctgtggacaa gtacaacgtg
     1981 agcggcacca acgggacctg cctgctggcc agcatggggc tgcagctgaa cctcacctat
     2041 gagaggaagg acaacacgac ggtgacaagg cttctcaaca tcaaccccaa caagacctcg
     2101 gccagcggga gctgcggcgc ccacctggtg actctggagc tgcacagcga gggcaccacc
     2161 gtcctgctct tccagttcgg gatgaatgca agttctagcc ggtttttcct acaaggaatc
     2221 cagttgaata caattcttcc tgacgccaga gaccctgcct ttaaagctgc caacggctcc
     2281 ctgcgagcgc tgcaggccac agtcggcaat tcctacaagt gcaacgcgga ggagcacgtc
     2341 cgtgtcacga aggcgttttc agtcaatata ttcaaagtgt gggtccaggc tttcaaggtg
     2401 gaaggtggcc agtttggctc tgtggaggag tgtctgctgg acgagaacag cacgctgatc
     2461 cccatcgctg tgggtggtgc cctggcgggg ctggtcctca tcgtcctcat cgcctacctc
     2521 gtcggcagga agaggagtca cgcaggctac cagactatcc gcggcatggt gagcaagggc
     2581 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2641 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     2701 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2761 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     2821 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     2881 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2941 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     3001 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     3061 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     3121 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     3181 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     3241 accgccgccg ggatcactct cggcatggac gagctgtaca agcggaccgt ctaatctaga
     3301 taagtaatga tcataatcag ccatatcaca tctgtagagg ttttacttgc tttaaaaaac
     3361 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3421 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3481 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3541 gtctggatcc ccgggtaccg agctcgaatt cgtaatcatg tcatagctgt ttcctgtgtg
     3601 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3661 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3721 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3781 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3841 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3901 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3961 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4021 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4081 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4141 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4201 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4261 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4321 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4381 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4441 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4501 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4561 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4621 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4681 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4741 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4801 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4861 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4921 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4981 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5041 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5101 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5161 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5221 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5281 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5341 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5401 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5461 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5521 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5581 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5641 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5701 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     5761 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     5821 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     5881 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     5941 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     6001 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     6061 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     6121 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     6181 taaaacgacg gccagtgcca agttgggatc tttgcattgg cccacggctc tcaggatggg
     6241 gatgctcccc ttcagcaccc ggttcccctt ggaaactgat ggtcctggct ctgtggcatg
     6301 gcagtggcac tgtgaggagc cccctaccag cagcacacag tggggttggc actgccacgc
     6361 tccggatgcc gcgctctgat ccaaccccat aatcaaggga acccgaattg ccccatcatt
     6421 gcccccacca cccccatcct gccgggccct cacaccccac gctgccttct ggtgacattc
     6481 cccagcccaa acccacggct tcatggctac cgcggggcat ttcccattgc cgccccatta
     6541 tcagctctgc acacctcccg ctgtacccat gcctcgtggc tgcccttctt tgacgtataa
     6601 tcttctaatt aatacccggc cttgtcaaag tggagcacaa acgttaatta attccccagc
     6661 aggcaggtaa ttaacagtgt gactcccttt ttgctgcgag tggggctgat acagagagat
     6721 gtggcactat ggagcccacg gggtcctggc actgggtgcc cacggaggtc cccatgtgct
     6781 gcagtgtcac cgcctccgag gagacagtat tgtccctgcg gtgtccctgc agctcagctc
     6841 tgtccacagg gccacctcca gtttggaggg gacacaatgc agccccgatg caacccatcc
     6901 tcgcagcatc ccagggacaa agaccccact gcaagaccgc acacagggct gggtcccgct
     6961 cccctaatat ctacagtgct tcttgcatgg ccccttaatc aatgcagtta atcagcatgc
     7021 gctcatgcac cgctctggag ctgcaaagcc cctcgcagcg ctgctcacca acaccgcgca
     7081 ccgccccggc ccagcctgca gcacgcgctg caaacaggaa agaaacaaaa tattgcccaa
     7141 atgtaggcaa aggcattcgg ctgccttgac ctccgccggg ccgggccctg cctgactcag
     7201 ctccttactc agcgctcgct tcctccctcc ggctgccacc gccgcagcgc acaccctgac
     7261 aaagagtggc ccttaacggg ctctgaggtg cacccagcag tgcactcagc agtccaaggg
     7321 ccggcctgga ggtttgcacc gctacgtgct gacattagca ttgaacttgg ccctgggtag
     7381 tgctgcaggc cgggcggggt gggtgtagag agtgcagcgc gcgttgcacc cggtgcccct
     7441 tcccctccct tgcatcccag caggctgcac cccagcacca ggcccgtgca tgcatgctcc
     7501 tggtgttatt gcagcctggt gcatgcatgc gtcttgatgg tgcagcgctg tgcatgcatc
     7561 ctccttggtg tgtagcagtc tagtgcatgc atacccctcg gtgttattgc tgctctgtgc
     7621 acgcacgctc attgtatcac ttcatcccag tgcatgcact cacactggag cgattgctgc
     7681 tcggtgcacg cacactcatt gtatcactgc agctcagtgg ctgcacgcac accggtgtta
     7741 ttgctgctcg gtgcgtgcat gcacatcagt gtcgctgcag ctcagtgcat gcacgctcat
     7801 tgcccatcgc tatccctgcc tctcctgctg gcgctccccg ggaggtgact tcaaggggac
     7861 cgcaggacca cctcgggggt ggggggaggg ctgcacacgc ggaccccgct ccccctcccc
     7921 aacaaagcac tgtggaatca aaaagggggg aggggggatg gaggggcgcg tcacaccccc
     7981 gccccacacc ctcacc
//
