LOCUS       sequence-435164-        6691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV SNAP-Synapsin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6691)
  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 6691)
  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 SNAP-Synapsin
FEATURES             Location/Qualifiers
     source          1..6691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        70..427
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        70..261
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(93..113)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      278..413
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             462..1256
                     /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             462..1256
                     /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(516..535)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1126..1145
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1444..1463)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1488..1535
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1864..2452
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2353..2372
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2627..2930
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2931..3134
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3085..3105
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3181..3726
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     polyA_signal    6045..6166
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6082..6101)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6136..6155
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6173..6628)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6310..6331)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6522..6541
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(6655..6691,1..68)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
       61 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
      121 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
      181 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
      241 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
      301 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
      361 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
      421 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
      481 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
      541 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
      601 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
      661 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
      721 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
      781 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
      841 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
      901 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
      961 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     1021 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     1081 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     1141 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     1201 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     1261 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     1321 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     1381 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     1441 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     1501 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     1561 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     1621 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     1681 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     1741 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     1801 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1861 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1921 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1981 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     2041 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2101 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2161 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2221 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2281 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2341 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2401 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2461 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2521 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
     2581 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     2641 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     2701 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     2761 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     2821 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     2881 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     2941 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     3001 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     3061 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     3121 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg tgaattcacc
     3181 atggacaaag actgcgaaat gaagcgcacc accctggata gccctctggg caagctggaa
     3241 ctgtctgggt gcgaacaggg cctgcaccgt atcatcttcc tgggcaaagg aacatctgcc
     3301 gccgacgccg tggaagtgcc tgccccagcc gccgtgctgg gcggaccaga gccactgatg
     3361 caggccaccg cctggctcaa cgcctacttt caccagcctg aggccatcga ggagttccct
     3421 gtgccagccc tgcaccaccc agtgttccag caggagagct ttacccgcca ggtgctgtgg
     3481 aaactgctga aagtggtgaa gttcggagag gtcatcagct acagccacct ggccgccctg
     3541 gccggcaatc ccgccgccac cgccgccgtg aaaaccgccc tgagcggaaa tcccgtgccc
     3601 attctgatcc cctgccaccg ggtggtgcag ggcgacctgg acgtgggggg ctacgagggc
     3661 gggctcgccg tgaaagagtg gctgctggcc cacgagggcc acagactggg caagcctggg
     3721 ctgggtcctg caggcggatc tcgagaagct gccaccatga actacctgcg gcgccgcctg
     3781 tcggacagca acttcatggc caatctgcct aatgggtata tgacagacct gcagcgtccg
     3841 caaccacctc cgccgccgcc ctcagccgcc agcccagggg ccactcccgg atccgctgct
     3901 gcctctgctg agagggcctc cacagccgct ccagtggcct ctccagcagc ccctagtccc
     3961 gggtcctcgg ggggcggtgg cttcttctcc tcgctgtcta acgcggtcaa acagaccaca
     4021 gcagctgcag ccgccacctt cagtgagcag gtgggcggtg gctctggggg cgcaggccgc
     4081 gggggcgccg ccgccagggt gctgctggtc atcgacgagc cgcacaccga ctgggcaaaa
     4141 tacttcaaag ggaagaagat ccatggagaa attgacatta aagtagagca agctgaattc
     4201 tccgatctca atcttgtggc tcatgccaat ggtggattct ccgtggacat ggaagttctt
     4261 cggaatgggg tcaaagttgt gaggtctctg aagccagact ttgtgctgat ccgccagcat
     4321 gccttcagca tggcacgtaa tggagactac cgcagtttgg tcattgggct gcagtatgct
     4381 gggatcccca gtgttaactc tttgcattct gtctacaact tttgtgacaa accctgggtg
     4441 tttgcccaga tggttcgact acacaagaag cttggaacag aggaattccc tctgattgat
     4501 cagactttct atcccaatca taaagagatg ctcagcagca caacataccc tgtagttgtg
     4561 aagatgggcc acgcacactc tgggatgggc aaggtcaagg tagacaacca acatgacttc
     4621 caggatattg caagtgttgt ggcactgact aagacatatg ccactgctga gcccttcatt
     4681 gatgctaaat acgatgtgcg tgtccagaag attgggcaga actacaaggc ctacatgagg
     4741 acatcagtgt cagggaactg gaagaccaat acaggctctg ctatgcttga gcagattgct
     4801 atgtctgaca ggtacaagtt gtgggtagac acgtgctcag agatttttgg gggacttgac
     4861 atctgcgcag tggaagcact gcatggcaag gacggaaggg atcacattat tgaggtggtg
     4921 ggctcctcca tgccactcat tggggatcac caggatgaag acaagcagct catcgtggaa
     4981 cttgtggtca acaagatgac tcaggctctg cctcggcagc gggatgcttc ccctggcagg
     5041 ggttcccaca gccagactcc atccccagga gccctgccct tgggccgcca gacctcccag
     5101 cagcctgcag gacctcctgc tcaacaacga cccccacccc agggaggccc tccacaacca
     5161 ggcccaggac ctcagcgcca gggacccccg ctgcaacagc gcccaccccc acaaggccag
     5221 caacatcttt ctggccttgg acccccagct ggcagccctc tgccccagcg cctaccaagt
     5281 cccacagcag cacctcagca gtccgcctct caggccacac caatgaccca gggtcaaggc
     5341 cgccagtcac ggccagtggc aggaggcccc ggagcacctc cagcagcccg cccgccggcc
     5401 tccccatctc cacagcgtca ggcagggccc ccacaggcta cccgtcaggc atctatctct
     5461 ggtccagctc caccgaaggt ctcaggagcc tcacccggag ggcagcagcg ccaaggccct
     5521 ccccagaaac ccccaggccc tgctggtccc attcgtcagg ccagccaggc aggtcccgga
     5581 cctcgcactg ggccacccac cacacagcag ccccggccca gcggcccagg tcctgctgga
     5641 cgtcccacca aaccacagct ggctcagaaa cccagccagg atgtgccacc acccatcatt
     5701 gctgctgccg ggggaccccc gcacccccag ctcaacaaat cccagtctct gaccaatgcc
     5761 ttcaaccttc cagagccagc cccgcccagg cccagcctta gccaggatga ggtgaaagct
     5821 gagaccatcc gcagcctgag gaagtctttc gccagcctct tctccgactg agagctcaag
     5881 cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga
     5941 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     6001 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     6061 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6121 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     6181 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     6241 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     6301 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     6361 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     6421 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     6481 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     6541 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     6601 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     6661 acccctattt gtttattttt ctaaatacat t
//
