LOCUS       sequence-488315-        7989 bp    DNA     circular SYN 03-OCT-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 7989)
  TITLE     Martin lab plasmids
REFERENCE   2  (bases 1 to 7989)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 3, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: EGFP Munc13-4
FEATURES             Location/Qualifiers
     source          1..7989
                     /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             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4717..4838
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4754..4773)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4808..4827
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4845..5300)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4982..5003)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5194..5213
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5327..5431
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5433..5790
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5433..5624
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5456..5476)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5641..5776
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5825..6619
                     /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             5825..6619
                     /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(5879..5898)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6489..6508
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6807..6826)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6851..6898
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7227..7815
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7716..7735
                     /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 gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgacatggcg
     1321 acactcctct cccatccgca gcagcgccct cccttcttgc gccaggccat caagataagg
     1381 cgccgcagag tcagagatct acaggatccc ccgccccaaa tggccccgga gatccagcct
     1441 ccatcccacc acttctcccc cgagcagcgg gccctgctct acgaggacgc actctacact
     1501 gtcttgcacc gcctgggtca tcctgagccc aaccatgtga cggaggcctc tgagctgctg
     1561 cgatacctgc aggaggcctt ccacgtggag cccgaggagc accagcagac actgcagcgg
     1621 gtcagggagc ttgagaagcc aatattttgt ctgaaggcaa cagtgaaaca ggccaagggc
     1681 attctgggca aagatgtcag tgggttcagc gacccctact gcctgctggg cattgagcag
     1741 ggggtaggtg tgccaggggg cagccccggg tcccggcatc ggcagaaggc tgtggtgagg
     1801 cacaccatcc ccgaggagga gacccaccgc acgcaggtca tcacccagac actcaacccc
     1861 gtctgggacg agaccttcat cctggagttt gaggacatca ccaatgcgag ctttcatctg
     1921 gacatgtggg acctggacac tgtggagtct gtccgacaga agcttgggga gctcacggat
     1981 ctgcatgggc ttcgcaggat ctttaaagag gcccggaagg acaaaggcca ggacgacttt
     2041 ctggggaacg tggttctgag gctgcaggac ctgcgctgcc gagaggacca gtggtacccc
     2101 ctggaacccc gcactgagac ctacccagac cgaggccagt gccacctcca gttccaactc
     2161 atccataagc ggagagccac ttcggccagc cgctcgcagc ccagctacac cgtgcacctc
     2221 cacctcctgc agcagcttgt gtcccacgag gtcacccagc acgaggcggg aagcacctcc
     2281 tgggacgggt cgctgagtcc ccaggctgcc accgtcctct ttctgcacgc cacacagaag
     2341 gacctatccg acttccacca gtccatggcg cagtggctgg cctacagccg cctctaccag
     2401 agcctggagt tccccagcag ctgcctcctg caccccatca ccagcatcga gtaccagtgg
     2461 atccagggtc ggctcaaggc agaacagcag gaggagctgg ccgcctcatt cagctccctg
     2521 ctgacctacg gcctctccct catccggagg ttccgctctg tcttccccct ctctgtctcg
     2581 gactccccag cccggctgca gtctcttctc agggtcctgg tacagatgtg caagatgaag
     2641 gcctttggag aactgtgccc caacaccgcc ccattgcccc agctggtgac tgaggccctg
     2701 cagactggca ccactgaatg gttccacctg aagcagcagc accatcaacc catggtgcag
     2761 ggcatcccgg aggcaggcaa ggccttgctg ggcctggtac aggatgtcat tggcgacctg
     2821 caccagtgcc agcgcacatg ggacaagatc ttccacaata ccctcaagat ccacctcttc
     2881 tccatggctt tccgggagct gcagtggctg gtggccaagc gggtgcagga ccacacgacg
     2941 gttgtgggtg atgtagtgtc cccagagatg ggcgagagtc tgttccagct ctacatcagc
     3001 ctcaaggagc tctgccagct gcgcatgagc tcctcagaga gggatggagt cctggccctg
     3061 gataatttcc accgctggtt ccagccggcc atcccctcct ggctgcagaa gacgtacaac
     3121 gaggccctgg cgcgggtgca gcgcgctgtg cagatggatg agctggtgcc cctgggtgaa
     3181 ctgaccaagc acagcacatc agcggtggat ctatccacct gctttgccca gatcagccac
     3241 actgcccggc agctggactg gccagaccca gaggaggcct tcatgattac cgtcaagttt
     3301 gtggaggaca cctgtcgcct ggccctggtg tactgcagcc ttataaaggc ccgggcccgc
     3361 gagctctctt caggccagaa ggaccaaggc caggcagcca acatgctgtg tgtggtggtg
     3421 aatgacatgg agcagctgcg gctggtgatc ggcaagttgc ccgcccagct ggcatgggag
     3481 gccctggagc agcgggtagg ggccgtgctg gagcaggggc agctgcagaa cacgctgcat
     3541 gcccagctgc agagcgcgct ggccgggctg ggccatgaga tccgcactgg cgtccgcacc
     3601 ctggccgagc agttggaggt gggcatcgcc aagcacatcc agaaactggt gggcgtcagg
     3661 gagtctgtcc tgcctgagga tgccattctg cccctgatga agttcctgga ggtggagctt
     3721 tgctacatga acaccaactt ggtgcaggaa aacttcagca gcctcctgac cctgctctgg
     3781 acccacacac tcacagtgct ggtggaggcg gccgcctccc agcgcagctc atccctggct
     3841 tccaacaggc tgaagattgc cctgcagaac ctggagatct gcttccacgc tgagggctgt
     3901 ggcctgccac ccgaggccct gcacactgcc accttccagg ctctgcagag ggacctggag
     3961 ctgcaggcgg cctccagccg ggaactcatc cggaagtact tctgcagccg aatccagcag
     4021 caggcagaaa ccacctctga ggagctgggg gctgtgacag tcaaggcctc ctaccgcgcc
     4081 tctgagcaga agctgcgtgt ggagctgctc agcgcctcca gcctgctgcc cctggactcc
     4141 aatggctcca gcgacccctt tgtccagctg accttggagc ccaggcatga gttccctgag
     4201 ctggccgccc gggagaccca gaagcacaag aaggaccttc acccattgtt tgatgagacc
     4261 tttgaattcc tggtgcctgc tgagccgtgc cgcaaggctg gggcatgcct cctgctcacc
     4321 gtgctggact acgacacgct gggggccgac gacctggaag gcgaggcctt cctgccgctg
     4381 cgtgaggtgc ccgggctgag tggctctgag gagcctggtg aggtgcctca gacccgcctg
     4441 cccctcacgt accccgcacc caacggggac ccaatcctgc agctgctgga gggccggaag
     4501 ggtgaccgag aggcccaggt ctttgtgagg ctgcggcggc accgggccaa gcaggcctcc
     4561 cagcatgcct tgcggccggc accgtaggga tccaccggat ctagataact gatcataatc
     4621 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     4681 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     4741 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4801 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa
     4861 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4921 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     4981 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     5041 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     5101 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     5161 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     5221 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     5281 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5341 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5401 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     5461 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5521 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     5581 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5641 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5701 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     5761 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     5821 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     5881 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     5941 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     6001 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     6061 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     6121 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     6181 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     6241 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     6301 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     6361 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     6421 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     6481 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     6541 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     6601 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     6661 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     6721 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     6781 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     6841 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     6901 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     6961 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     7021 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     7081 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     7141 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     7201 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     7261 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     7321 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     7381 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7441 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     7501 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     7561 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     7621 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7681 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7741 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7801 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7861 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     7921 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     7981 ggagttccg
//
