LOCUS       sequence-494529-        7992 bp    DNA     circular SYN 03-DEC-2025
DEFINITION  Distribution and localization of human wild-type Munc13-4 in cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7992)
  TITLE     Martin lab plasmids
REFERENCE   2  (bases 1 to 7992)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 3, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: mCherry shRNA Resistant Munc13-4
FEATURES             Location/Qualifiers
     source          1..7992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             553..1260
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(698..716)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     966..985
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4720..4841
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4757..4776)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4811..4830
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4848..5303)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4985..5006)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5197..5216
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5330..5434
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5436..5793
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5436..5627
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5459..5479)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5644..5779
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5828..6622
                     /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             5828..6622
                     /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(5882..5901)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6492..6511
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6810..6829)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6854..6901
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7230..7818
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7719..7738
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
      601 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      661 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
      721 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
      781 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
      841 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      901 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
      961 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     1021 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     1081 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1141 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     1201 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1261 tccggactca gatctcgagc tcaaatggcg acactcctct cccatccgca gcagcgccct
     1321 cccttcttgc gccaggccat caagataagg cgccgcagag tcagagatct acaggatccc
     1381 ccgccccaaa tggccccgga gatccagcct ccatcccacc acttctcccc cgagcagcgg
     1441 gccctgctct acgaggacgc actctacact gtcttgcacc gcctgggtca tcctgagccc
     1501 aaccatgtga cggaggcctc tgagctgctg cgatacctgc aggaggcctt ccacgtggag
     1561 cccgaggagc accagcagac actgcagcgg gtcagggagc ttgagaagcc aatattttgt
     1621 ctgaaggcaa cagtgaaaca ggccaagggc attctgggca aagatgtcag tgggttcagc
     1681 gacccctact gcctgctggg cattgagcag ggggtaggtg tgccaggggg cagccccggg
     1741 tcccggcatc ggcagaaggc tgtggtgagg cacaccatcc ccgaggagga gacccaccgc
     1801 acgcaggtca tcacccagac actcaacccc gtctgggacg agaccttcat cctggagttt
     1861 gaggacatca ccaatgcgag ctttcatctg gacatgtggg acctggacac tgtggagtct
     1921 gtccgacaga agcttgggga gctcacggat ctgcatgggc ttcgcaggat ctttaaagag
     1981 gcccggaagg acaaaggcca ggacgacttt ctggggaacg tggttctgag gctgcaggac
     2041 ctgcgctgcc gagaggacca gtggtacccc ctggaacccc gcactgagac ctacccagac
     2101 cgaggccagt gccacctcca gttccaactc atccataagc ggagagccac ttcggccagc
     2161 cgctcgcagc ccagctacac cgtgcacctc cacctcctgc agcagcttgt gtcccacgag
     2221 gtcacccagc acgaggcggg aagcacctcc tgggacgggt cgctgagtcc ccaggctgcc
     2281 accgtcctct ttctgcacgc cacacagaag gacctatccg acttccacca gtccatggcg
     2341 cagtggctgg cctacagccg cctctaccag agcctggagt tccccagcag ctgcctcctg
     2401 caccccatca ccagcatcga gtaccagtgg atccagggtc ggctcaaggc agaacagcag
     2461 gaggagctgg ccgcctcatt cagctccctg ctgacctacg gcctctccct catccggagg
     2521 ttccgctctg tcttccccct ctctgtctcg gactccccag cccggctgca gtctcttctc
     2581 agggtcctgg tacagatgtg caagatgaag gcctttggag aactgtgccc caacaccgcc
     2641 ccattgcccc agctggtgac tgaggccctg cagactggca ccactgaatg gttccacctg
     2701 aagcagcagc accatcaacc catggtgcag ggcatcccgg aggcaggcaa ggccttgctg
     2761 ggcctggtac aggatgtcat tggcgacctg caccagtgcc agcgcacatg ggacaagatc
     2821 ttccacaata ccctcaagat ccacctcttc tccatggctt tccgggagct gcagtggctg
     2881 gtggccaagc gggtgcagga ccacacgacg gttgtgggtg atgtagtgtc cccagagatg
     2941 ggcgagagtc tgttccagct ctacatcagc ctcaaggagc tctgccagct gcgcatgagc
     3001 tcctcagaga gggatggagt cctggccctg gataatttcc accgctggtt ccagccggcc
     3061 atcccctcct ggctgcagaa gacgtacaac gaggccctgg cgcgggtgca gcgcgctgtg
     3121 cagatggatg agctggtgcc cctgggtgaa ctgaccaagc acagcacatc cgcagtagac
     3181 ctctcaacct gctttgccca gatcagccac actgcccggc agctggactg gccagaccca
     3241 gaggaggcct tcatgattac cgtcaagttt gtggaggaca cctgtcgcct ggccctggtg
     3301 tactgcagcc ttataaaggc ccgggcccgc gagctctctt caggccagaa ggaccaaggc
     3361 caggcagcca acatgctgtg tgtggtggtg aatgacatgg agcagctgcg gctggtgatc
     3421 ggcaagttgc ccgcccagct ggcatgggag gccctggagc agcgggtagg ggccgtgctg
     3481 gagcaggggc agctgcagaa cacgctgcat gcccagctgc agagcgcgct ggccgggctg
     3541 ggccatgaga tccgcactgg cgtccgcacc ctggccgagc agttggaggt gggcatcgcc
     3601 aagcacatcc agaaactggt gggcgtcagg gagtctgtcc tgcctgagga tgccattctg
     3661 cccctgatga agttcctgga ggtggagctt tgctacatga acaccaactt ggtgcaggaa
     3721 aacttcagca gcctcctgac cctgctctgg acccacacac tcacagtgct ggtggaggcg
     3781 gccgcctccc agcgcagctc atccctggct tccaacaggc tgaagattgc cctgcagaac
     3841 ctggagatct gcttccacgc tgagggctgt ggcctgccac ccgaggccct gcacactgcc
     3901 accttccagg ctctgcagag ggacctggag ctgcaggcgg cctccagccg ggaactcatc
     3961 cggaagtact tctgcagccg aatccagcag caggcagaaa ccacctctga ggagctgggg
     4021 gctgtgacag tcaaggcctc ctaccgcgcc tctgagcaga agctgcgtgt ggagctgctc
     4081 agcgcctcca gcctgctgcc cctggactcc aatggctcca gcgacccctt tgtccagctg
     4141 accttggagc ccaggcatga gttccctgag ctggccgccc gggagaccca gaagcacaag
     4201 aaggaccttc acccattgtt tgatgagacc tttgaattcc tggtgcctgc tgagccgtgc
     4261 cgcaaggctg gggcatgcct cctgctcacc gtgctggact acgacacgct gggggccgac
     4321 gacctggaag gcgaggcctt cctgccgctg cgtgaggtgc ccgggctgag tggctctgag
     4381 gagcctggtg aggtgcctca gacccgcctg cccctcacgt accccgcacc caacggggac
     4441 ccaatcctgc agctgctgga gggccggaag ggtgaccgag aggcccaggt ctttgtgagg
     4501 ctgcggcggc accgggccaa gcaggcctcc cagcatgcct tgcggccggc accgtaatcg
     4561 aattctgcag tcgacggtac cgcgggcccg ggatccaccg gatctagata actgatcata
     4621 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     4681 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     4741 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4801 cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt
     4861 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     4921 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     4981 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     5041 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     5101 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     5161 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     5221 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     5281 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5341 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5401 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     5461 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5521 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     5581 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5641 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5701 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     5761 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     5821 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     5881 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     5941 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     6001 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     6061 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     6121 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     6181 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     6241 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     6301 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     6361 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     6421 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     6481 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     6541 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     6601 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     6661 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     6721 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     6781 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     6841 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     6901 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     6961 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     7021 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     7081 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     7141 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7201 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7261 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7321 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     7381 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7441 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7501 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7561 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7621 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7681 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7741 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7801 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7861 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     7921 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     7981 tatggagttc cg
//
