LOCUS       sequence-488338-        8203 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 8203)
  TITLE     Martin lab plasmids
REFERENCE   2  (bases 1 to 8203)
  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: emiRFP 670 shRNA Resistant Munc13-4
FEATURES             Location/Qualifiers
     source          1..8203
                     /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      548..557
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4931..5052
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4968..4987)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5022..5041
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5059..5514)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5196..5217)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5408..5427
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5541..5645
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5647..6004
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5647..5838
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5670..5690)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5855..5990
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6039..6833
                     /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             6039..6833
                     /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(6093..6112)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6703..6722
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7021..7040)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7065..7112
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7441..8029
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7930..7949
                     /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 ccggttcgcc accatggcgg aaggctccgt cgccaggcag cctgacctct tgacctgcga
      601 acatgaagag atccacctcg ccggctcgat ccagccgcat ggcgcgcttc tggtcgtcag
      661 cgaacatgat catcgcgtca tccaggccag cgccaacgcc gcggaatttc tgaatctcgg
      721 aagcgtactc ggcgttccgc tcgccgagat cgacggcgat ctgttgatca agatcctgcc
      781 gcatctcgat cccaccgccg aaggcatgcc ggtcgcggtg cgctgccgga tcggcaatcc
      841 ctctacggag tactgcggtc tgatgcatcg gcctccggaa ggcgggctga tcatcgaact
      901 cgaacgtgcc ggcccgtcga tcgatctgtc aggcacgctg gcgccggcgc tggagcggat
      961 ccgcacggcg ggttcactgc gcgcgctgtg cgatgacacc gtgctgctgt ttcagcagtg
     1021 caccggctac gaccgggtga tggtgtatcg tttcgatgag caaggccacg gcctggtatt
     1081 ctccgagtgc catgtgcctg ggctcgaatc ctatttcggc aaccgctatc cgtcgtcgac
     1141 tgtcccgcag atggcgcggc agctgtacgt gcggcagcgc gtccgcgtgc tggtcgacgt
     1201 cacctatcag ccggtgccgc tggagccgcg gctgtcgccg ctgaccgggc gcgatctcga
     1261 catgtcgggc tgcttcctgc gctcgatgtc gccgtgccat ctgcagttcc tgaaggacat
     1321 gggcgtgcgc gccaccctgg cggtgtcgct ggtggtcggc ggcaagctgt ggggcctggt
     1381 tgtctgtcac cattatctgc cgcgcttcat ccgtttcgag ctgcgggcga tctgcaaacg
     1441 gctcgccgaa aggatcgcga cgcggatcac cgcgcttgag agctccggac tcagatctcg
     1501 agctcaagct tcgaattctg cagtcgacat ggcgacactc ctctcccatc cgcagcagcg
     1561 ccctcccttc ttgcgccagg ccatcaagat aaggcgccgc agagtcagag atctacagga
     1621 tcccccgccc caaatggccc cggagatcca gcctccatcc caccacttct cccccgagca
     1681 gcgggccctg ctctacgagg acgcactcta cactgtcttg caccgcctgg gtcatcctga
     1741 gcccaaccat gtgacggagg cctctgagct gctgcgatac ctgcaggagg ccttccacgt
     1801 ggagcccgag gagcaccagc agacactgca gcgggtcagg gagcttgaga agccaatatt
     1861 ttgtctgaag gcaacagtga aacaggccaa gggcattctg ggcaaagatg tcagtgggtt
     1921 cagcgacccc tactgcctgc tgggcattga gcagggggta ggtgtgccag ggggcagccc
     1981 cgggtcccgg catcggcaga aggctgtggt gaggcacacc atccccgagg aggagaccca
     2041 ccgcacgcag gtcatcaccc agacactcaa ccccgtctgg gacgagacct tcatcctgga
     2101 gtttgaggac atcaccaatg cgagctttca tctggacatg tgggacctgg acactgtgga
     2161 gtctgtccga cagaagcttg gggagctcac ggatctgcat gggcttcgca ggatctttaa
     2221 agaggcccgg aaggacaaag gccaggacga ctttctgggg aacgtggttc tgaggctgca
     2281 ggacctgcgc tgccgagagg accagtggta ccccctggaa ccccgcactg agacctaccc
     2341 agaccgaggc cagtgccacc tccagttcca actcatccat aagcggagag ccacttcggc
     2401 cagccgctcg cagcccagct acaccgtgca cctccacctc ctgcagcagc ttgtgtccca
     2461 cgaggtcacc cagcacgagg cgggaagcac ctcctgggac gggtcgctga gtccccaggc
     2521 tgccaccgtc ctctttctgc acgccacaca gaaggaccta tccgacttcc accagtccat
     2581 ggcgcagtgg ctggcctaca gccgcctcta ccagagcctg gagttcccca gcagctgcct
     2641 cctgcacccc atcaccagca tcgagtacca gtggatccag ggtcggctca aggcagaaca
     2701 gcaggaggag ctggccgcct cattcagctc cctgctgacc tacggcctct ccctcatccg
     2761 gaggttccgc tctgtcttcc ccctctctgt ctcggactcc ccagcccggc tgcagtctct
     2821 tctcagggtc ctggtacaga tgtgcaagat gaaggccttt ggagaactgt gccccaacac
     2881 cgccccattg ccccagctgg tgactgaggc cctgcagact ggcaccactg aatggttcca
     2941 cctgaagcag cagcaccatc aacccatggt gcagggcatc ccggaggcag gcaaggcctt
     3001 gctgggcctg gtacaggatg tcattggcga cctgcaccag tgccagcgca catgggacaa
     3061 gatcttccac aataccctca agatccacct cttctccatg gctttccggg agctgcagtg
     3121 gctggtggcc aagcgggtgc aggaccacac gacggttgtg ggtgatgtag tgtccccaga
     3181 gatgggcgag agtctgttcc agctctacat cagcctcaag gagctctgcc agctgcgcat
     3241 gagctcctca gagagggatg gagtcctggc cctggataat ttccaccgct ggttccagcc
     3301 ggccatcccc tcctggctgc agaagacgta caacgaggcc ctggcgcggg tgcagcgcgc
     3361 tgtgcagatg gatgagctgg tgcccctggg tgaactgacc aagcacagca catcagcggt
     3421 ggatctatcc acctgctttg cccagatcag ccacactgcc cggcagctgg actggccaga
     3481 cccagaggag gccttcatga ttaccgtcaa gtttgtggag gacacctgtc gcctggccct
     3541 ggtgtactgc agccttataa aggcccgggc ccgcgagctc tcttcaggcc agaaggacca
     3601 aggccaggca gccaacatgc tgtgtgtggt ggtgaatgac atggagcagc tgcggctggt
     3661 gatcggcaag ttgcccgccc agctggcatg ggaggccctg gagcagcggg taggggccgt
     3721 gctggagcag gggcagctgc agaacacgct gcatgcccag ctgcagagcg cgctggccgg
     3781 gctgggccat gagatccgca ctggcgtccg caccctggcc gagcagttgg aggtgggcat
     3841 cgccaagcac atccagaaac tggtgggcgt cagggagtct gtcctgcctg aggatgccat
     3901 tctgcccctg atgaagttcc tggaggtgga gctttgctac atgaacacca acttggtgca
     3961 ggaaaacttc agcagcctcc tgaccctgct ctggacccac acactcacag tgctggtgga
     4021 ggcggccgcc tcccagcgca gctcatccct ggcttccaac aggctgaaga ttgccctgca
     4081 gaacctggag atctgcttcc acgctgaggg ctgtggcctg ccacccgagg ccctgcacac
     4141 tgccaccttc caggctctgc agagggacct ggagctgcag gcggcctcca gccgggaact
     4201 catccggaag tacttctgca gccgaatcca gcagcaggca gaaaccacct ctgaggagct
     4261 gggggctgtg acagtcaagg cctcctaccg cgcctctgag cagaagctgc gtgtggagct
     4321 gctcagcgcc tccagcctgc tgcccctgga ctccaatggc tccagcgacc cctttgtcca
     4381 gctgaccttg gagcccaggc atgagttccc tgagctggcc gcccgggaga cccagaagca
     4441 caagaaggac cttcacccat tgtttgatga gacctttgaa ttcctggtgc ctgctgagcc
     4501 gtgccgcaag gctggggcat gcctcctgct caccgtgctg gactacgaca cgctgggggc
     4561 cgacgacctg gaaggcgagg ccttcctgcc gctgcgtgag gtgcccgggc tgagtggctc
     4621 tgaggagcct ggtgaggtgc ctcagacccg cctgcccctc acgtaccccg cacccaacgg
     4681 ggacccaatc ctgcagctgc tggagggccg gaagggtgac cgagaggccc aggtctttgt
     4741 gaggctgcgg cggcaccggg ccaagcaggc ctcccagcat gccttgcggc cggcaccgta
     4801 gggatccacc ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt
     4861 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     4921 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4981 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5041 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     5101 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     5161 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     5221 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     5281 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     5341 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     5401 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     5461 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     5521 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5581 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5641 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     5701 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     5761 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5821 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5881 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5941 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     6001 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     6061 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     6121 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     6181 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     6241 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     6301 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     6361 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     6421 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     6481 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     6541 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     6601 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     6661 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     6721 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     6781 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     6841 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6901 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6961 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     7021 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     7081 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     7141 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     7201 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     7261 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     7321 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     7381 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     7441 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     7501 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7561 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     7621 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7681 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7741 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7801 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7861 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7921 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7981 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     8041 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     8101 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
     8161 caattacggg gtcattagtt catagcccat atatggagtt ccg
//
