LOCUS       sequence-435144-        6218 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PGK mScarlet-Synaptophysin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6218)
  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 6218)
  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: PGK mScarlet-Synaptophysin
FEATURES             Location/Qualifiers
     source          1..6218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        129..639
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     558..576
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     regulatory      655..664
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             661..1356
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(794..812)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1062..1081
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2339..2927
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2392..2412)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2810..2821)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2930..2946)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2931..2947)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     misc_feature    2943..2995
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3119..3240
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3156..3175)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3210..3229
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3247..3702)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3384..3405)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3596..3615
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3729..3833
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3835..4192
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3835..4026
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3858..3878)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4043..4178
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4227..5021
                     /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             4227..5021
                     /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(4281..4300)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4891..4910
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5209..5228)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5253..5300
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5629..6217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6118..6137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
       61 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
      121 taatccacgg ggttggggtt gcgccttttc caaggcagcc ctgggtttgc gcagggacgc
      181 ggctgctctg ggcgtggttc cgggaaacgc agcggcgccg accctgggtc tcgcacattc
      241 ttcacgtccg ttcgcagcgt cacccggatc ttcgccgcta cccttgtggg ccccccggcg
      301 acgcttcctg ctccgcccct aagtcgggaa ggttccttgc ggttcgcggc gtgccggacg
      361 tgacaaacgg aagccgcacg tctcactagt accctcgcag acggacagcg ccagggagca
      421 atggcagcgc gccgaccgcg atgggctgtg gccaatagcg gctgctcagc agggcgcgcc
      481 gagagcagcg gccgggaagg ggcggtgcgg gaggcggggt gtggggcggt agtgtgggcc
      541 ctgttcctgc ccgcgcggtg ttccgcattc tgcaagcctc cggagcgcac gtcggcagtc
      601 ggctccctcg ttgaccgaat caccgacctc tctccccagg tggatccacc ggtcgccacc
      661 atggtgagca agggcgaggc agtgatcaag gagttcatgc ggttcaaggt gcacatggag
      721 ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
      781 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt ctcctgggac
      841 atcctgtccc ctcagttcat gtacggctcc agggccttca ccaagcaccc cgccgacatc
      901 cccgactact ataagcagtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
      961 gaggacggcg gcgccgtgac cgtgacccag gacacctccc tggaggacgg caccctgatc
     1021 tacaaggtga agctccgcgg caccaacttc cctcctgacg gccccgtaat gcagaagaag
     1081 acaatgggct gggaagcgtc caccgagcgg ttgtaccccg aggacggcgt gctgaagggc
     1141 gacattaaga tggccctgcg cctgaaggac ggcggccgct acctggcgga cttcaagacc
     1201 acctacaagg ccaagaagcc cgtgcagatg cccggcgcct acaacgtcga ccgcaagttg
     1261 gacatcacct cccacaacga ggactacacc gtggtggaac agtacgaacg ctccgagggc
     1321 cgccactcca ccggcggcat ggacgagctg tacaagtccg gactcagatc catggacgtg
     1381 gtgaatcagc tggtggctgg gggtcagttc cgggtggtca aggagcccct tggcttcgtg
     1441 aaggtgctgc agtgggtctt tgccatcttc gcctttgcta cgtgtggcag ctacaccggg
     1501 gagcttcggc tgagcgtgga gtgtgccaac aagacggaga gtgccctcaa catcgaagtt
     1561 gaattcgagt accccttcag gctgcaccaa gtgtactttg atgcaccctc ctgcgtcaaa
     1621 gggggcacta ccaagatctt cctggttggg gactactcct cgtcggctga attctttgtc
     1681 accgtggctg tgtttgcctt cctctactcc atgggggccc tggccaccta catcttcctg
     1741 cagaacaagt accgagagaa caacaaaggg cctatgatgg actttctggc tacagccgtg
     1801 ttcgctttca tgtggctagt tagttcatca gcctgggcca aaggcctgtc cgatgtgaag
     1861 atggccacgg acccagagaa cattatcaag gagatgccca tgtgccgcca gacagggaac
     1921 acatgcaagg aactgaggga ccctgtgact tcaggactca acacctcagt ggtgtttggc
     1981 ttcctgaacc tggtgctctg ggttggcaac ttatggttcg tgttcaagga gacaggctgg
     2041 gcagccccat tcatgcgcgc acctccaggc gccccggaaa agcaaccagc acctggcgat
     2101 gcctacggcg atgcgggcta cgggcagggc cccggaggct atgggcccca ggactcctac
     2161 gggcctcagg gtggttatca acccgattac gggcagccag ccagcggtgg cggtggctac
     2221 gggcctcagg gcgactatgg gcagcaaggc tatggccaac agggtgcgcc cacctccttc
     2281 tccaatcaga tgtaaggtac cgcgggcccg ggatcaattc gatatcaagc ttatcgataa
     2341 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     2401 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     2461 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     2521 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     2581 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     2641 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2701 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     2761 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     2821 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     2881 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg
     2941 acctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg gatccaccgg
     3001 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     3061 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     3121 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3181 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3241 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3301 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3361 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3421 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3481 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3541 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3601 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     3661 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     3721 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3781 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     3841 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3901 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3961 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4021 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4081 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4141 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     4201 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4261 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4321 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4381 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     4441 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4501 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4561 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4621 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4681 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4741 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4801 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4861 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4921 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4981 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     5041 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5101 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5161 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     5221 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     5281 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     5341 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     5401 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     5461 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5521 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5581 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5641 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5701 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5761 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5821 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5881 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5941 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6001 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6061 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaa
//
