LOCUS       sequence-222083-        6562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express in mammalian cells the endocytic protein Rab5 fused to 
            mCherry by transfection or by generating AAV virions..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6562)
  AUTHORS   Pantelis Tsoulfas
  TITLE     Eukaryotic expression plasmids for expression and visualization of 
            early endocytic markers Rab5 and Rab7
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6562)
  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            "Unpublished"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: pAAV-UbC-mCherry Rab5
FEATURES             Location/Qualifiers
     source          1..6562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        152..551
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1121..1140
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     primer_bind     1837..1856
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1907..2611
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDVLYK"
     primer_bind     complement(2049..2067)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2317..2336
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3309..3785
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3508..3527)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3825..3965
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3825..3954
                     /label=AAV2 ITR
     rep_origin      4040..4495
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4127..4146)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4337..4358
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4512..4531)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4631..4653
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4691..4709)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4777..4881
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4882..4950,4951..5742)
                     /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     complement(5100..5119)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5913..6501
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6402..6421
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtttaaac tggcctccgc gccgggtttt ggcgcctccc
      181 gcgggcgccc ccctcctcac ggcgagcgct gccacgtcag acgaagggcg cagcgagcgt
      241 cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta
      301 gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg
      361 gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg
      421 gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca
      481 cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat
      541 cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc
      601 tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa
      661 ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt
      721 gaatagaacc ttcgctaatg cgggaaagct cttattcggg tgagatgggc tggggcacca
      781 tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt cgtctgttgc
      841 gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc
      901 cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg gggccacctg
      961 ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg cctagggtag
     1021 gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt gaggcgtcag
     1081 tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg gttttgaact
     1141 atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa
     1201 tcatttgggt caatatgtaa ttttcagtgt tagactagta aattgtccgc taaattctgg
     1261 ccgtttttgg cttttttgtt agacgaagct aaggcgcgcc tgagaacttc agggtgagtt
     1321 tggggaccct tgattgttct ttctttttcg ctattgtaaa attcatgtta tatggagggg
     1381 gcaaagtttt cagggtgttg tttagaatgg gaagatgtcc cttgtatcac catgcatgga
     1441 ccctcatgat aattttgttt ctttcacttt ctactctgtt gacaaccatt gtctcctctt
     1501 attttctttt cattttctgt aactttttcg ttaaacttta gcttgcattt gtaacgaatc
     1561 tagacaatca gggtatatta tattgtactt cagcacagtt ttagagaaca attgttataa
     1621 ttaaatgata aggtagaata tttctgcata taaattctgg ctggcgtgga aatattctta
     1681 ttggtagaaa caactacatc ctggtcatca tcctgccttt ctctttatgg ttacaatgat
     1741 atacactgtt tgagatgagg ataaaatact ctgagtccaa accgggcccc tctgctaacc
     1801 atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt
     1861 ctcatcaagc ttggatccga attcgctagc gatctagcga attcatgtga gcaagggcga
     1921 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
     1981 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     2041 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     2101 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     2161 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     2221 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     2281 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     2341 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     2401 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     2461 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     2521 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     2581 ctccaccggc ggcatggacg tgctgtacaa gcttatggct agtcgaggcg caacaagacc
     2641 caacgggcca aatacgggaa ataaaatatg ccagttcaaa ctagtacttc tgggagagtc
     2701 cgctgttggc aaatcaagcc tagtgcttcg ttttgtgaaa ggccaatttc atgaatttca
     2761 agagagtacc attggggctg cttttctaac ccaaactgta tgtcttgatg acactacagt
     2821 aaagtttgaa atatgggata cagctggtca agaacgatac catagcctag caccaatgta
     2881 ctacagagga gcacaagcag ccatagttgt atatgatatc acaaatgagg agtcctttgc
     2941 aagagcaaaa aattgggtta aagaacttca gaggcaagca agtcctaaca ttgtaatagc
     3001 tttatcggga aacaaggccg acctagcaaa taaaagagca gtagatttcc aggaagcaca
     3061 gtcctatgca gatgacaata gtttattatt catggagaca tccgctaaaa catcaatgaa
     3121 tgtaaatgaa atattcatgg caatagctaa aaaattgcca aagaatgaac cacaaaatcc
     3181 aggagcaaat tctgccagag gaagaggagt agaccttacc gaacccacac aaccaaccag
     3241 gaatcagtgt tgtagtaact aagcggccat ctcgagtcta gagttaacgg ccatattggc
     3301 caatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     3361 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     3421 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     3481 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     3541 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     3601 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     3661 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     3721 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     3781 tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     3841 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     3901 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     3961 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4021 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4081 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     4141 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4201 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     4261 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4321 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     4381 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     4441 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     4501 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4561 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4621 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4681 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4741 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4801 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4861 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4921 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4981 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5041 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5101 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5161 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5221 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5281 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5341 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5401 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5461 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5521 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5581 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5641 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5701 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5761 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5821 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5881 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5941 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6001 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6061 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6121 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6181 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6241 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6301 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6361 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6421 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6481 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6541 tgctggcctt ttgctcacat gt
//
