LOCUS       sequence-486092-        5336 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Mammalian expression of N-terminal FLAG-tagged rat Rab3A wild type.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5336)
  TITLE     Martin lab plasmids 2025 (Yamaga)
REFERENCE   2  (bases 1 to 5336)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pFLAG-CMV2-Rab3A
FEATURES             Location/Qualifiers
     source          1..5336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             935..958
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1677..2299
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1876..1895)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2323..2343)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2328..2657
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2508..2643
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2695..2714)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2696..2714)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2728..2744)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2728..2744)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2741..2763)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2752..2768
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2776..2782,2783..2800,2801..2806))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2821..2842
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2959..2976)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3130..3718)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3210..3229)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3889..4680,4681..4749))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4512..4531
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4750..4854)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4881..5309
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4968..4987)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5178..5199
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attgatctac catggactac aaagacgatg acgacaagct
      961 tgcggccgcg aattcatcga tagatctgat atcggtacca gtcgacatgg cctcagccac
     1021 agactctcga tatgggcaga aggagtcctc agaccagaac ttcgactata tgttcaagat
     1081 cctgatcatt ggtaacagca gtgtgggcaa aacctcattc cttttccgct acgcagacga
     1141 ctcttttact ccagcctttg tcagcactgt gggcatagac ttcaaggtca aaaccatcta
     1201 ccgcaatgac aagaggatca agctgcagat ctgggacaca gcaggacaag agcggtaccg
     1261 aaccatcacc acagcctact accggggcgc catgggcttc attctaatgt atgacatcac
     1321 caatgaggag tccttcaatg cagtgcagga ctggtccact cagatcaaaa cttactcatg
     1381 ggacaatgcc caggtgctgc tggtggggaa caagtgcgac atggaggacg agcgagtggt
     1441 gtcctcagaa cgaggccggc agctggccga ccacctgggc tttgagttct ttgaggccag
     1501 cgccaaggac aacattaatg tcaagcagac ctttgaacgt ctggtggacg tgatctgtga
     1561 gaagatgtcg gagtccctag atactgcaga ccctgcagtc acgggtgcca agcagggccc
     1621 acagctcacc gaccagcagg caccacctca tcaggattgc gcctgctgag gatcccgggt
     1681 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
     1741 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
     1801 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
     1861 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
     1921 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
     1981 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
     2041 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
     2101 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
     2161 taaaataact ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc
     2221 aaccggtggg acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc
     2281 agtagatgcc tgttgaattg ggtacgcggc cagcttggct gtggaatgtg tgtcagttag
     2341 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2401 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2461 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     2521 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     2581 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     2641 gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt aattccctat
     2701 agtgagtcgt attaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc
     2761 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     2821 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2881 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2941 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3001 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3061 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3121 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3181 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3241 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3301 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3361 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3421 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3481 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3541 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3601 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3661 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3721 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3781 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3841 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3901 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3961 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4021 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4081 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4141 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4201 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4261 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4321 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4381 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4441 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4501 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4561 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4621 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4681 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4741 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4801 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4861 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4921 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4981 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5041 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5101 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5161 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5221 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     5281 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caattt
//
