LOCUS       sequence-198613-        5482 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and linker-mEGFP sequence for C-terminus tagging of 
            human LAMP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5482)
  AUTHORS   Allen Institute for Cell Science
  TITLE     Allen Institute for Cell Science Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5482)
  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: 
            "Unpublished"
COMMENT     Sequence Label: AICSDP-19:LAMP1-mEGFP
FEATURES             Location/Qualifiers
     source          1..5482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(376..1089,1090..1092))
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(421..442)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     770..789
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1029..1050
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2204..2220)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2204..2220)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2217..2239)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2228..2244
                     /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(2252..2258,2259..2276,2277..2282))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2297..2318
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2435..2452)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2606..3194)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2686..2705)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3365..4156,4157..4225))
                     /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     3988..4007
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4226..4330)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4398..4416
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4454..4476)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4576..4595
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4789..4811
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4803..4820
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4804..4820
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ccctcagaga aagcggcacc cccaccccct cagagagagc ggcaccccca cccctcagag
       61 acagcggcac ccccacccct cagagacagc ggcaccccca ccccctcaga gagagcaccc
      121 ccaccctcaa aatcctactt tggtaaatat taatgtttaa ccagttagca aaatatttaa
      181 cccagttagc aaaattaaca ccgtcatttc gatagttact ctgttgtgca tagtaaatgc
      241 tgcaagatga acttcacatt ggatgaagca gatttgagaa acgtttgcca gaaagtgtgc
      301 ctccccttcg acaggaccct aagcccagag aaaggaacag aggcccctgc agctgctgtg
      361 gctgcgtgca ccaggctact tgtacagctc gtccatgccg agagtgatcc cggcggcggt
      421 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga
      481 ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
      541 ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa
      601 gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta
      661 gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag
      721 ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt
      781 gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt
      841 gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt
      901 cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt
      961 cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg
     1021 gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc
     1081 gcccttgctc acggtcgcca ccggcggatc cgcgccggtg ctgccggtgc tgccggtgct
     1141 gccaaattcg atagtctgat agcctgcgtg actcctcttc ctgccgacga ggtaggcgat
     1201 gaggacgatg aggaccagcc ccgccagggc accacccaca gcgatgggga tcagcatgct
     1261 gttctcgtcc agcagacact cctccactgc caagacagac ggatggtgtc agggctctgc
     1321 tcacacaggg gcctgcaaac aggcctctct ccccaccact cccccacatc agccaaaagg
     1381 caaagaggaa ccctggcgga aggttccagc tctagctgga attgtcactt ggggaagccg
     1441 ctgcagagac tggtgctctc tgcacggcga ggaaaccaca cacgtctctc ccccgtctgg
     1501 aatcaggggc tgtggccgga cagctgtgct gcgtccaggg cctctgtgca catcagatga
     1561 cagtgttgta ttcaacgttc atgggagaag cgccaccact gtttcttaaa atgattccct
     1621 tactctcaaa tggtaccagg ggctcccagc tgagagacac gctgggcctc tctaaagcag
     1681 ggggagttac ctctagaatc tcagaccggc aagaagaaac agctccttgc agaggaacag
     1741 gggcgtcgcc ccagcccaca cggaagcgac taaacgtcca caggcggagt ctgaagcacg
     1801 tcctccacac cccgcgacag ctgccctcgg tgacactcac cagagccaaa ctggccacct
     1861 tccaccttga aagcctggac ccacactttg aatatattga ctgaaaacgc cttcgtgaca
     1921 cggacgtgct cctccgcgtt gcacttgtag gaattgccga ctgtggcctg cagcgctcgc
     1981 agggagccgt tggcagcttt aaaggcaggg tctgcaacac acacagaggt ctagtggctc
     2041 cagaataccc tgggcgcgcg ggcgggggcg gggctcgcag agattgtggg ttctcacctc
     2101 tggcgtcagg aagaattgta ttcaactgga ttccttgtag gaaaaaccga tcccaatacg
     2161 cgtcaattca tcccaatggc gcgccgagct tggcgtaatc atggtcatag ctgtttcctg
     2221 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     2281 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     2341 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     2401 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2461 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     2521 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2581 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2641 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2701 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2761 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2821 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2881 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2941 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3001 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     3061 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3121 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3181 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3241 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3301 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3361 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3421 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3481 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     3541 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     3601 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     3661 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     3721 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     3781 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     3841 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     3901 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     3961 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4021 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4081 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4141 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4201 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4261 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4321 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     4381 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     4441 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     4501 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     4561 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     4621 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     4681 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     4741 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     4801 gttgtaaaac gacggccagt gaattgacgc gtattgggat gctcggcgcg ccattgggat
     4861 gaggcttttt attacattgt tacaggcaat gcattacgtg agctggtccc gtgtacaatc
     4921 cccaagccac ggcaaaacat caggagaacg caccgatctc acattgtaca cagcgcagaa
     4981 caggatcacc ccgaatgtca acagaagacc aaaaatcact ttacaaataa aaataaaaat
     5041 aacacccatt tttatattta aaaaagtgcc agcccttgag ctgcagacat tagagacgcc
     5101 caaggacgtg ccgccactgg aacgaggccg ccagccacgt gtggcccgtc ctggccttta
     5161 aagctgaact gaagcgctcc agacactcat ctctggagtt cctctgtctt aggatgttcc
     5221 ctctctactg ttgtaatgtg tgagcatgtc agcctcacca cgagtgacct tcatgcgtcc
     5281 tacgtgtgac ggccagaggc tttccctttg taaggcttaa atctgagtgc agtaaagaat
     5341 gaagaaacca aacaccagaa ccctaaactc aaatgatatg gagccaattt agagcggcac
     5401 ccccacccct cagagacagc ggcattccac ccctcggaga gagcagcacc cccaccccct
     5461 cagagagagc ggcaccccca cc
//
