LOCUS       sequence-488320-        8009 bp    DNA     circular SYN 03-OCT-2025
DEFINITION  Proximity labeling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8009)
  TITLE     Martin lab plasmids
REFERENCE   2  (bases 1 to 8009)
  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: Flag Apex2 Munc13-4
FEATURES             Location/Qualifiers
     source          1..8009
                     /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      537..546
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             549..572
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    4737..4858
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4774..4793)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4828..4847
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4865..5320)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5002..5023)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5214..5233
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5347..5451
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5453..5810
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5453..5644
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5476..5496)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5661..5796
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5845..6639
                     /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             5845..6639
                     /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(5899..5918)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6509..6528
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6827..6846)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6871..6918
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7247..7835
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7736..7755
                     /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 gctagcgcca
      541 ccatggtgga ctacaaggat gacgacgata aggggaaatc atacccaaca gtgtccgcag
      601 actaccagga tgccgtggag aaagccaaga agaagctgag agggtttatc gcagaaaaga
      661 ggtgcgcacc tctgatgctg agactggctt tccacagcgc aggcaccttt gacaagggaa
      721 ccaaaacagg cggacccttt ggaacaatca agcaccctgc tgaactggca catagtgcca
      781 acaatgggct ggacatcgcc gtgcggctgc tggaacctct gaaagcagag ttcccaattc
      841 tgtcctacgc cgatttttat cagctggcag gagtggtcgc tgtggaggtc actgggggcc
      901 ccaaggtgcc tttccaccca ggacgggagg acaaaccaga accacctcca gaggggcgcc
      961 tgccagatcc gacaaagggc tccgaccatc tgcgagatgt gtttgggaaa gctatgggcc
     1021 tgactgacca ggatatcgtc gcactgtctg gagggcacac cattggcgcc gctcataagg
     1081 aaaggtcagg cttcgaggga ccctggacaa gcaaccccct gattttcgac aattcttact
     1141 ttactgaact gctgagtgga gagaaggaag ggctgctgca gctgcccagc gacaaagccc
     1201 tgctgtccga tcccgtgttc agacctctgg tcgataagta tgcagccgac gaggatgctt
     1261 ttttcgcaga ttacgcagaa gcacatcaga agctgtcaga actgggattt gccgacgcct
     1321 ctggctcggg cagcatggcg acactcctct cccatccgca gcagcgccct cccttcttgc
     1381 gccaggccat caagataagg cgccgcagag tcagagatct acaggatccc ccgccccaaa
     1441 tggccccgga gatccagcct ccatcccacc acttctcccc cgagcagcgg gccctgctct
     1501 acgaggacgc actctacact gtcttgcacc gcctgggtca tcctgagccc aaccatgtga
     1561 cggaggcctc tgagctgctg cgatacctgc aggaggcctt ccacgtggag cccgaggagc
     1621 accagcagac actgcagcgg gtcagggagc ttgagaagcc aatattttgt ctgaaggcaa
     1681 cagtgaaaca ggccaagggc attctgggca aagatgtcag tgggttcagc gacccctact
     1741 gcctgctggg cattgagcag ggggtaggtg tgccaggggg cagccccggg tcccggcatc
     1801 ggcagaaggc tgtggtgagg cacaccatcc ccgaggagga gacccaccgc acgcaggtca
     1861 tcacccagac actcaacccc gtctgggacg agaccttcat cctggagttt gaggacatca
     1921 ccaatgcgag ctttcatctg gacatgtggg acctggacac tgtggagtct gtccgacaga
     1981 agcttgggga gctcacggat ctgcatgggc ttcgcaggat ctttaaagag gcccggaagg
     2041 acaaaggcca ggacgacttt ctggggaacg tggttctgag gctgcaggac ctgcgctgcc
     2101 gagaggacca gtggtacccc ctggaacccc gcactgagac ctacccagac cgaggccagt
     2161 gccacctcca gttccaactc atccataagc ggagagccac ttcggccagc cgctcgcagc
     2221 ccagctacac cgtgcacctc cacctcctgc agcagcttgt gtcccacgag gtcacccagc
     2281 acgaggcggg aagcacctcc tgggacgggt cgctgagtcc ccaggctgcc accgtcctct
     2341 ttctgcacgc cacacagaag gacctatccg acttccacca gtccatggcg cagtggctgg
     2401 cctacagccg cctctaccag agcctggagt tccccagcag ctgcctcctg caccccatca
     2461 ccagcatcga gtaccagtgg atccagggtc ggctcaaggc agaacagcag gaggagctgg
     2521 ccgcctcatt cagctccctg ctgacctacg gcctctccct catccggagg ttccgctctg
     2581 tcttccccct ctctgtctcg gactccccag cccggctgca gtctcttctc agggtcctgg
     2641 tacagatgtg caagatgaag gcctttggag aactgtgccc caacaccgcc ccattgcccc
     2701 agctggtgac tgaggccctg cagactggca ccactgaatg gttccacctg aagcagcagc
     2761 accatcaacc catggtgcag ggcatcccgg aggcaggcaa ggccttgctg ggcctggtac
     2821 aggatgtcat tggcgacctg caccagtgcc agcgcacatg ggacaagatc ttccacaata
     2881 ccctcaagat ccacctcttc tccatggctt tccgggagct gcagtggctg gtggccaagc
     2941 gggtgcagga ccacacgacg gttgtgggtg atgtagtgtc cccagagatg ggcgagagtc
     3001 tgttccagct ctacatcagc ctcaaggagc tctgccagct gcgcatgagc tcctcagaga
     3061 gggatggagt cctggccctg gataatttcc accgctggtt ccagccggcc atcccctcct
     3121 ggctgcagaa gacgtacaac gaggccctgg cgcgggtgca gcgcgctgtg cagatggatg
     3181 agctggtgcc cctgggtgaa ctgaccaagc acagcacatc agcggtggat ctatccacct
     3241 gctttgccca gatcagccac actgcccggc agctggactg gccagaccca gaggaggcct
     3301 tcatgattac cgtcaagttt gtggaggaca cctgtcgcct ggccctggtg tactgcagcc
     3361 ttataaaggc ccgggcccgc gagctctctt caggccagaa ggaccaaggc caggcagcca
     3421 acatgctgtg tgtggtggtg aatgacatgg agcagctgcg gctggtgatc ggcaagttgc
     3481 ccgcccagct ggcatgggag gccctggagc agcgggtagg ggccgtgctg gagcaggggc
     3541 agctgcagaa cacgctgcat gcccagctgc agagcgcgct ggccgggctg ggccatgaga
     3601 tccgcactgg cgtccgcacc ctggccgagc agttggaggt gggcatcgcc aagcacatcc
     3661 agaaactggt gggcgtcagg gagtctgtcc tgcctgagga tgccattctg cccctgatga
     3721 agttcctgga ggtggagctt tgctacatga acaccaactt ggtgcaggaa aacttcagca
     3781 gcctcctgac cctgctctgg acccacacac tcacagtgct ggtggaggcg gccgcctccc
     3841 agcgcagctc atccctggct tccaacaggc tgaagattgc cctgcagaac ctggagatct
     3901 gcttccacgc tgagggctgt ggcctgccac ccgaggccct gcacactgcc accttccagg
     3961 ctctgcagag ggacctggag ctgcaggcgg cctccagccg ggaactcatc cggaagtact
     4021 tctgcagccg aatccagcag caggcagaaa ccacctctga ggagctgggg gctgtgacag
     4081 tcaaggcctc ctaccgcgcc tctgagcaga agctgcgtgt ggagctgctc agcgcctcca
     4141 gcctgctgcc cctggactcc aatggctcca gcgacccctt tgtccagctg accttggagc
     4201 ccaggcatga gttccctgag ctggccgccc gggagaccca gaagcacaag aaggaccttc
     4261 acccattgtt tgatgagacc tttgaattcc tggtgcctgc tgagccgtgc cgcaaggctg
     4321 gggcatgcct cctgctcacc gtgctggact acgacacgct gggggccgac gacctggaag
     4381 gcgaggcctt cctgccgctg cgtgaggtgc ccgggctgag tggctctgag gagcctggtg
     4441 aggtgcctca gacccgcctg cccctcacgt accccgcacc caacggggac ccaatcctgc
     4501 agctgctgga gggccggaag ggtgaccgag aggcccaggt ctttgtgagg ctgcggcggc
     4561 accgggccaa gcaggcctcc cagcatgcct tgcggccggc accgtaggga tccaccggat
     4621 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     4681 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     4741 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4801 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac
     4861 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     4921 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     4981 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     5041 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     5101 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     5161 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     5221 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     5281 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     5341 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5401 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     5461 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     5521 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     5581 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5641 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5701 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     5761 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     5821 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     5881 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5941 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     6001 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     6061 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     6121 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     6181 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     6241 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     6301 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     6361 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     6421 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     6481 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     6541 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     6601 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     6661 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     6721 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     6781 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     6841 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     6901 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     6961 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     7021 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     7081 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     7141 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     7201 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     7261 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     7321 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     7381 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     7441 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     7501 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     7561 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     7621 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     7681 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     7741 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     7801 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     7861 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     7921 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
     7981 ttagttcata gcccatatat ggagttccg
//
