LOCUS       sequence-441071-        6343 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBI-BFP-NL1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6343)
  AUTHORS   Dou D, Aiken J, Holzbaur ELF
  TITLE     RAB3 phosphorylation by pathogenic LRRK2 impairs trafficking of 
            synaptic vesicle precursors.
  JOURNAL   J Cell Biol. 2024 Jun 3;223(6):e202307092. doi: 
            10.1083/jcb.202307092. Epub 2024 Mar 21.
  PUBMED    38512027
REFERENCE   2  (bases 1 to 6343)
  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: "J Cell 
            Biol."; date: "2024-06-3"; volume: "223(6):e202307092. doi"; pages: 
            " 10.1083/jcb.202307092. Epub 2024 Mar 21"
COMMENT     Sequence Label: pBI-BFP-NL1
FEATURES             Location/Qualifiers
     source          1..6343
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             133..834
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    960..1041
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1074..1178
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1179..1247,1248..2039)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1397..1416)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2210..2798
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2699..2718
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2952..2969
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    2980..3061
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      3201..3210
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      5144..5153
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5549..5575)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(5811..6014)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5840..5860)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6015..6318
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ctaatgaccc cgtaattgat tactattaat aactactcga gtatgtcgag gtggcgtgta
       61 cggtgggagg cctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagaatt
      121 ccggtcgcca ccatgagcga gctgattaag gagaacatgc acatgaagct gtacatggag
      181 ggcaccgtgg acaaccatca cttcaagtgc acatccgagg gcgaaggcaa gccctacgag
      241 ggcacccaga ccatgagaat caaggtggtc gagggcggcc ctctcccctt cgccttcgac
      301 atcctggcta ctagcttcct ctacggcagc aagaccttca tcaaccacac ccagggcatc
      361 cccgacttct tcaagcagtc cttccctgag ggcttcacat gggagagagt caccacatac
      421 gaagacgggg gcgtgctgac cgctacccag gacaccagcc tccaggacgg ctgcctcatc
      481 tacaacgtca agatcagagg ggtgaacttc acatccaacg gccctgtgat gcagaagaaa
      541 acactcggct gggaggcctt caccgagacg ctgtaccccg ctgacggcgg cctggaaggc
      601 agaaacgaca tggccctgaa gctcgtgggc gggagccatc tgatcgcaaa catcaagacc
      661 acatatagat ccaagaaacc cgctaagaac ctcaagatgc ctggcgtcta ctatgtggac
      721 tacagactgg aaagaatcaa ggaggccaac aacgagacct acgtcgagca gcacgaggtg
      781 gcagtggcca gatactgcga cctccctagc aaactggggc acaagcttaa ttaaagcggc
      841 cgcgactcta gaggatcata atcagccata ccacatttgt agaggtttta cttgctttaa
      901 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
      961 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     1021 ataaagcatt tttttcactg ccgacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     1081 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1141 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     1201 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     1261 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     1321 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     1381 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     1441 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     1501 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     1561 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     1621 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     1681 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     1741 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     1801 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     1861 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     1921 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     1981 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     2041 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     2101 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     2161 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2221 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2281 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2341 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2401 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2461 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2521 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2581 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2641 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2701 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2761 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2821 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     2881 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     2941 cgaccgagcg cagcgagtca gtgagcgagg aagctcgggg cagtgaaaaa aatgctttat
     3001 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     3061 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     3121 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcctc tggagatggg
     3181 cccgacgtcg catgctcccg gccgccatgg ccgcgggatt aatcagtttg tctctctcat
     3241 ctggctatac cctggttgtt gaatgtgaat gggggtgtgg gtggggatgg ggcagtgtat
     3301 tattctgtcc tccagtaaat gtgttgaatg tatgtaaggg ctgaatccct ggtatagtgt
     3361 tgggaatcat tgtgatggtg ttaggtgtca ttagtggaat atcatcaggt gacctcctca
     3421 tagctagagt ataatctggg ggacaggcgg tccgaagaac cacctcatgt ggatggatgg
     3481 actcacactc gtgatccaag tcagtgtgct tcatttggag agacataatt tcctcttctg
     3541 gagcatgggt taggtcgttg gtggtcgtgc gctgagggct gcacctccgg tggacatcat
     3601 gtctcctctt atccttcttg tagtacaggg ctgcaaaagc caagatgttg agaaacagca
     3661 gagaggcccc cactgcgata gtgacactta gctctgtgga gtagtccctc tgatccaccg
     3721 agaaggggct tggttgttgc ttgggatcat cctgtttggc agtgggaaag gctgatgtga
     3781 ctggtgtgga attcttcctt gtaggtctga gagtgatgtc cgtggatggc actttagttg
     3841 ttgtcgaggt atactgagaa atgtcattga gattatgcag atgaggtacc agctccagcc
     3901 agagatttac cttattggct ctgtaatgct ctttaactct cggttttaat ccaatatgga
     3961 gataaagctg gtctttctgg gaatatctgg tccatgctac ttcttcaaag cggttgggtt
     4021 tggtatggat gaattttgtg tcttgaggaa ctggttgatt tgggtcacca gttttagcaa
     4081 aattcgtcca gtatgtcatt actactgcac tcaacatcac atcattcttg gagaaattgc
     4141 aaggaaataa ctctgtaggg ccaatcatgg ggattcccaa cacataggga acctcatccc
     4201 catgagctgc atcagcccaa gctggaactt gatctgtttg gcaatgatga taaaaggcat
     4261 agaagtatgt aggtgagcca aagttcgagt gaaggtctgc tgtggccaca gcaggtgcta
     4321 cccattgatg gtccgtaaac aaagccaaca atgtcttcct tctagtttca ggattatggc
     4381 gatcagccca gtcagtatac atgaatttaa tggtttctct caaaacatct ttgccttcag
     4441 gatatccata taaattatca acaaaattag aaacagcaaa gtcgaaatca ctggctgata
     4501 caccatcatc actatctact atattttcga caaacttcaa cccttcccct tggttaactc
     4561 ccaacattat atcatagttg aggaactctc cttgttccat cagtatctga gggtcatctg
     4621 gtattacatc accatcgatc acaggtccaa aggctatgtg gtatcgggct ggttgaacat
     4681 cttgatcaac aagttctttg taaggcttct tctgcaggca ttctactaac tctactgtat
     4741 ctgaaacatt gcagccaact tttgtggcca gaattctagc gtattttgca ggctggaaac
     4801 taacagccca gctggaaagg gctgttccac tctgagctat tgctcgttga aaaagtcctt
     4861 cagaataatg ggataaagtc agcaggttga cacatgaacc cccagcgcct gatccaaaca
     4921 cagtgattcg caaggggtca ccaccaaaga acccaatgtt ctcgctggtc catcttaggg
     4981 cctggatgag gtcaaggagc ccgtagtttc ctttggcagc ctgatcccct gtgctcaaga
     5041 agccaagtac cccaagccga tagttgactg tgatgacgat cacattgcca tagcttgcca
     5101 agacactccc atcatacaga tttccagtac cttccatgta agagccgcca tggatgtaca
     5161 ccatcactgg tttgggaccc ccactgtccc gaatatcatc ttcagttggg acatagatgt
     5221 ttaagtatag acagtcttca ctctggtctt ggacgtatga tgaaaccaca tccaagttat
     5281 tagtgaacca cacaggaagc ataacttcag gcaatctgcc atcaatgata ttctggggac
     5341 atacaggagc aaactgagtg gcgttccgga tgtcagacca gggagatggt ggttctggag
     5401 gctggaaacg atgttctcct gttggtggag cggcatatgg aaccccaaga aactgaatga
     5461 caggacccaa aatttcatta ttgagttctt tcttaattcc cctaatcttg ccaaagttag
     5521 tagtaaccaa tgggtctaca tcatccaatg cgtagtccgg aacgtcgtat ggatactttt
     5581 gagagagtac gtgaaaagtc tgtagcaaac atcccaacaa gcagagagtc aatggggcac
     5641 cggatcccct gtggaccaca catgccatca tagctctcca aacataattt ggccacatgc
     5701 atctgggaag tgccatggtc ccaagcttga gctcgagatc tgagtccggt agcgctagca
     5761 cgcgtcagct gactagagga tccctagcgg atctgacggt tcactaaacc agctctgctt
     5821 atatagacct cccaccgtac acgcctaccg cccatttgcg tcaatggggc ggagttgtta
     5881 cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg
     5941 gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt gatgtactgc
     6001 caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga
     6061 aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg
     6121 tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac
     6181 cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca
     6241 tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt
     6301 accgtaagtt atgtaacgcg gaactccata tatgggctat gaa
//
