LOCUS       sequence-487947-        4752 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Protein expression and purification of rat wild-type Rab3a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4752)
  TITLE     Martin lab plasmids
REFERENCE   2  (bases 1 to 4752)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: twin strep HRV3C Flag Rab3a
FEATURES             Location/Qualifiers
     source          1..4752
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        301..504
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     455..475
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      543..552
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(555..560,561..584,585..620,621..644)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     CDS             663..686
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             687..710
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1548..1669
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1585..1604)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1639..1658
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1676..2131)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1813..1834)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2025..2044
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2158..2262
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2287..2307)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2331..2541
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2353..2549
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2400..2535
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2584..3378
                     /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             2584..3378
                     /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(2638..2657)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3248..3267
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3566..3585)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3610..3657
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3986..4574
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4475..4494
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(4749..4752,1..300)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
       61 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      121 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      181 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      241 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      301 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      361 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      421 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      481 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      541 tcgccaccat gggcagcgca tggagtcatc ctcaattcga gaaaggtgga ggttctggcg
      601 gtggatcggg aggttcagcg tggagccacc cgcagttcga aaaatccgga ggtgcttctg
      661 gtctggaagt tctgttccag ggacccgatt acaaggatga cgacgataag tccggactca
      721 gatctcgagc tcaagcttcg aattctgcag tcgacatggc ctcagccaca gactctcgat
      781 atgggcagaa ggagtcctca gaccagaact tcgactatat gttcaagatc ctgatcattg
      841 gtaacagcag tgtgggcaaa acctcattcc ttttccgcta cgcagacgac tcttttactc
      901 cagcctttgt cagcactgtg ggcatagact tcaaggtcaa aaccatctac cgcaatgaca
      961 agaggatcaa gctgcagatc tgggacacag caggacaaga gcggtaccga accatcacca
     1021 cagcctacta ccggggcgcc atgggcttca ttctaatgta tgacatcacc aatgaggagt
     1081 ccttcaatgc agtgcaggac tggtccactc agatcaaaac ttactcatgg gacaatgccc
     1141 aggtgctgct ggtggggaac aagtgcgaca tggaggacga gcgagtggtg tcctcagaac
     1201 gaggccggca gctggccgac cacctgggct ttgagttctt tgaggccagc gccaaggaca
     1261 acattaatgt caagcagacc tttgaacgtc tggtggacgt gatctgtgag aagatgtcgg
     1321 agtccctaga tactgcagac cctgcagtca cgggtgccaa gcagggccca cagctcaccg
     1381 accagcaggc accacctcat caggattgcg cctgctgagg atccaccgga tctagataac
     1441 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     1501 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     1561 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     1621 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt
     1681 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     1741 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     1801 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     1861 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     1921 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     1981 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2041 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     2101 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     2161 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2221 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     2281 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2341 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     2401 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     2461 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     2521 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     2581 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     2641 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     2701 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     2761 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     2821 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     2881 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     2941 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3001 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3061 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     3121 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3181 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     3241 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     3301 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     3361 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     3421 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     3481 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     3541 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     3601 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     3661 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     3721 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     3781 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     3841 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3901 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3961 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4021 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4081 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4141 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4201 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4261 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4321 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4381 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4441 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4501 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4561 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4621 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4681 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     4741 gagttccgcg tt
//
