LOCUS       sequence-435172-        5011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV HA-RIM2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5011)
  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 5011)
  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: CMV HA-RIM2
FEATURES             Location/Qualifiers
     source          1..5011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..330
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        331..534
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     485..505
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     531..555
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          672..768
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             832..858
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2150..2284
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2174..2193)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2228..2247
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2395..2411)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2395..2411)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2408..2430)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2419..2435
                     /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(2443..2449,2450..2467,2468..2473))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2488..2509
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2626..2643)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2797..3385)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2877..2896)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3556..4347,4348..4416))
                     /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     4179..4198
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4417..4521)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4589..4607
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4645..4667)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4767..4786
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4980..5002
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4994..5011
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4995..5011
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gagttcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc
       61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      541 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
      601 cgggaccgat ccagcctccg gactctagag gatccggtac tagaggaact gaaaaaccag
      661 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg
      721 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa
      781 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg ggcccaccat gtacccatac
      841 gatgttccag attacgctct tatggccatg gaggcccgaa ttctaatgtc ggctccactc
      901 gggccccggg gccgcccggc tcccaccccg gcggcctctc agcctcctcc gcagcccgag
      961 atgccggacc tcagccacct cacggaagag gagaggaaaa tcatccaggc tgtcatggat
     1021 cgtcagaaga aagaagagga gaaggagcag tccgtgctca aaaagctgca tcaacaattt
     1081 gaaatgtata aggagcaagt caagaaaatg ggagaggaat cacaacagca gcaagagcag
     1141 aagggcgacg ccccgacctg cggcatctgc cacaagacaa aatttgcaga tggatgcggc
     1201 cataactgtt catattgcca aaccaagttc tgtgctcgtt gtggaggtcg agtgtcatta
     1261 cgctcaaaca aggttatgtg ggtgtgtaat ttgtgccgaa aacaacaaga aatcctcact
     1321 aagtcgggcg cgtggtttta caatagtggg tctaatacac cgcagcagcc tgatcaaaag
     1381 gctcttcgag ggcttcgcag tgaggaagcc cctcaggaga agaaggcaaa actgcatgag
     1441 cagacgcagt tccagggacc ccccggtgac tcatcagtac ctgcagttga gagaggtcga
     1501 gctcatgggc tcacaagaca ggattctatt aagaatggct caggaatgaa gcaccagatc
     1561 gccagtgaca tgccttcaga cagaaaaaga agtccgtcag tgtccaggga tcaaaatcga
     1621 agatacgacc aaagtgaaga aagagaggaa tattcacagt atgttccttc agatagcaca
     1681 atgcctagat ctccatcaga ttatgctgat agacgatctc aacgtgaacc tcaattttat
     1741 gaagaacctg atcatttaaa ttatagggat tctaacagga gaggccatag acattccaaa
     1801 gagtatattg tagacgacga agatgtggag agcagagatg aatatgaaag acaaaggaga
     1861 gaggaggagt accaggcacg ctacagaagt gatccaaatt tggcccggta tccggtaaag
     1921 ccacaaccct atgaagagca aatgcggatc cacgctgaag tgtcccgggc acgacacgag
     1981 agaaggcaca gtgatgtttc tttggcaaat gctgaactag aagattccag gatttctctg
     2041 ctaaggatgg atagaccatc aaggcaaaga tctgtatctg aacgtagagc tgcaatggaa
     2101 aaccagcgat cgtattcatg gaaataactc gaggtaccgc ggccgcgggg atccagacat
     2161 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2221 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2281 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     2341 tttttcggat cctctagagt cgatctgcag gcatgctagc ttggcgtaat catggtcata
     2401 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2461 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2521 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2581 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2641 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2701 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2761 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2821 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2881 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2941 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3001 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3061 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3121 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3181 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3241 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3301 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3361 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3421 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3481 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3541 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3601 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3661 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3721 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3781 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3841 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3901 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3961 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4021 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4081 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4141 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4201 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4261 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4321 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4381 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4441 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4501 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4561 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     4621 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     4681 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     4741 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     4801 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     4861 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     4921 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     4981 ccagtcacga cgttgtaaaa cgacggccag t
//
