LOCUS       sequence-463631-        4894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 3xFlag-tagged human Rab34 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4894)
  TITLE     expression plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4894)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: 3xFlag-Rab34
FEATURES             Location/Qualifiers
     source          1..4894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             81..146
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    946..1080
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(970..989)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1024..1043
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1193..1213)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1193..1211)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1232..1248)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1232..1248)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1245..1267)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1256..1272
                     /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(1280..1286,1287..1304,1305..1310))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1325..1346
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1463..1480)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1634..2222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1714..1733)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2393..3184,3185..3253))
                     /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     3016..3035
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3254..3358)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3384..3839)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3521..3542)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3733..3752
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3910..4894
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agacttgttc
       61 tttttgcagg atctaccatg gactacaaag accatgacgg tgattataaa gatcatgaca
      121 tcgattacaa ggatgacgat gacaagggat ccatgaacat tctggcaccc gtgcggaggg
      181 atcgcgtcct ggcggagctg ccccagtgcc tgaggaagga ggccgctttg cacgggcaca
      241 aagacttcca cccccgcgtc acctgcgcct gccaggagca ccggacaggc accgtgggat
      301 ttaagatctc caaggtcatt gtggtggggg acctgtcggt ggggaagact tgcctcatta
      361 ataggttctg caaagacacc tttgataaga attacaaggc caccattgga gtggacttcg
      421 agatggaacg atttgaggtg ctgggcattc ccttcagttt gcagctttgg gataccgctg
      481 ggcaggagag gttcaaatgc attgcatcaa cctactatag aggagctcaa gccatcatca
      541 ttgtcttcaa cctgaatgat gtggcatctc tggaacatac caagcagtgg ctggccgatg
      601 ccctgaagga gaatgaccct tccagtgtgc ttctcttcct tacccctgct cagtatgcgc
      661 tgatggagaa agacgccctc caggtggccc aggagatgaa ggctgagtac tgggcagtct
      721 catccctcac tggtgagaat gtccgagaat tcttcttccg tgtggcagca ctgacctttg
      781 aggccaatgt gctggctgag ctggagaaat cgggggctcg acgcattggg gatgttgtcc
      841 gcatcaacag tgatgacagc aacctctacc taactgccag caagaagaag cccacatgtt
      901 gcccatgact cgagcctcta gaactatagt gagtcgtatt acgtagatcc agacatgata
      961 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     1021 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     1081 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     1141 taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt tccctttagt
     1201 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1261 atccgctcac aattccacac aacatacgag ccgggagcat aaagtgtaaa gcctggggtg
     1321 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     1381 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1441 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1501 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1561 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1621 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1681 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1741 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1801 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1861 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1921 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1981 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2041 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2101 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2161 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2221 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2281 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2341 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2401 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2461 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     2521 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     2581 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     2641 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     2701 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     2761 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     2821 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     2881 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     2941 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3001 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3061 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3121 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3181 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3241 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3301 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3361 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt
     3421 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     3481 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     3541 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3601 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     3661 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     3721 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3781 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3841 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     3901 attacgccag tcgaccgcca attcaatatg gcgtatatgg actcatgcca attcaatatg
     3961 gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg
     4021 gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg
     4081 gcggacctgg cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg
     4141 aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg
     4201 gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca
     4261 atatggctat tggccaggtt caatactatg tattggccct atgccatata gtattccata
     4321 tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata
     4381 tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt ctctatatat
     4441 ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt
     4501 gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac
     4561 ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta
     4621 ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac
     4681 ttggcaagta cattactatt ggaagtacgc cagggtacat tggcagtact cccattgacg
     4741 tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg
     4801 aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg
     4861 ggaggtctat ataagcaatg ctcgtttagg gaac
//
