LOCUS       sequence-491006-        5234 bp    DNA     circular SYN 29-OCT-2025
DEFINITION  Mammalian expression of N-terminal Myc-tagged rat Rab3A F59S mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5234)
  TITLE     Martin lab plasmids 2025 (Yamaga)
REFERENCE   2  (bases 1 to 5234)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 29, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pEF-cMyc-Rab3A(F59S)
FEATURES             Location/Qualifiers
     source          1..5234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..1194
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          247..1185
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1142..1162
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1218..1227
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1227..1256
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    2094..2215
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2131..2150)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2185..2204
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2222..2677)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2359..2380)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2571..2590
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2704..2808
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2833..2853)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2877..3087
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2899..3095
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2946..3081
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3130..3924
                     /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             3130..3924
                     /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(3184..3203)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3794..3813
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4112..4131)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4156..4203
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4532..5120
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5021..5040
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcggccgcct cgtgaggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
       61 cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg
      121 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
      181 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
      241 acacaggtaa gtgccgtgtg tggttccggc gggcctggcc tctttacggg ttatggccct
      301 tgcgtgcctt gaattacttc cacctggcag cagtacgtga ttcttgatcc cgagcttcgg
      361 gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct
      421 tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc
      481 gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg
      541 acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatcagca cactggtatt
      601 tcggtttttg ggggcgcggg cggcgacggg ggccgtgcgt cccagcgcac atgttcggcg
      661 aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg
      721 cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg
      781 gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctggaggg
      841 tgctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg
      901 aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg
      961 tccaggcacc tcgattagtt gtcgagcttt tggagtacgt cgtctttagg ttggggggag
     1021 gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct
     1081 tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt
     1141 ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgaggaatt
     1201 ggctagcgct accggtcgcc accatggaac aaaaacttat ttcagaagaa gatttgtccg
     1261 gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga catggcctca gccacagact
     1321 ctcgatatgg gcagaaggag tcctcagacc agaacttcga ctatatgttc aagatcctga
     1381 tcattggtaa cagcagtgtg ggcaaaacct cattcctttt ccgctacgca gacgactctt
     1441 ttactccagc ctttgtcagc actgtgggca tagacagcaa ggtcaaaacc atctaccgca
     1501 atgacaagag gatcaagctg cagatctggg acacagcagg acaagagcgg taccgaacca
     1561 tcaccacagc ctactaccgg ggcgccatgg gcttcattct aatgtatgac atcaccaatg
     1621 aggagtcctt caatgcagtg caggactggt ccactcagat caaaacttac tcatgggaca
     1681 atgcccaggt gctgctggtg gggaacaagt gcgacatgga ggacgagcga gtggtgtcct
     1741 cagaacgagg ccggcagctg gccgaccacc tgggctttga gttctttgag gccagcgcca
     1801 aggacaacat taatgtcaag cagacctttg aacgtctggt ggacgtgatc tgtgagaaga
     1861 tgtcggagtc cctagatact gcagaccctg cagtcacggg tgccaagcag ggcccacagc
     1921 tcaccgacca gcaggcacca cctcatcagg attgcgcctg ctgaggatcc accggatcta
     1981 gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     2041 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2101 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2161 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg
     2221 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     2281 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     2341 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     2401 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     2461 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2521 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2581 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2641 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     2701 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2761 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     2821 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2881 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     2941 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3001 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3061 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     3121 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3181 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3241 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3301 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3361 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3421 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3481 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3541 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3601 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     3661 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3721 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3781 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3841 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     3901 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     3961 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4021 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4081 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     4141 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     4201 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     4261 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     4321 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     4381 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4441 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4501 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4561 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4621 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4681 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4741 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4801 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4861 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4921 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4981 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5041 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5101 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5161 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5221 ttaccgccat gcat
//
