LOCUS       sequence-394430-        5696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of eGFP-Rab31 (human) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5696)
  AUTHORS   Banworth MJ, Liang Z, Li G
  TITLE     A novel membrane targeting domain mediates the endosomal or Golgi 
            localization specificity of small GTPases Rab22 and Rab31.
  JOURNAL   J Biol Chem. 2022 Sep;298(9):102281. doi: 10.1016/j.jbc.2022.102281.
            Epub 2022 Jul 19.
  PUBMED    35863437
REFERENCE   2  (bases 1 to 5696)
  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: "J Biol 
            Chem."; date: "2022-09"; volume: "298(9):102281. doi"; pages: " 
            10.1016/j.jbc.2022.102281. Epub 2022 Jul 19"
COMMENT     Sequence Label: pBI-eGFP-Rab31
FEATURES             Location/Qualifiers
     source          1..5696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(136..154,155..177,178..196,197..219,220..238,239..261,
                     262..280,281..303,304..322,323..345,346..364,365..387,
                     388..406)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        439..477
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             join(616..618,619..621,622..1332)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(616..618,619..621,622..1332)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(661..682)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(922..941)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1269..1290
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          1971..2543
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2675..2694)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2740..2795
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2794..2813)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3193..3210)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3364..3952)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3444..3463)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4123..4914,4915..4983))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4746..4765
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4984..5088)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5156..5174
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    5537..5671
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5574..5593)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5628..5647
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ctgcaggaat tcggggccgc ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa
       61 acagcgtgga tggcgtctcc aggcgatctg acggttcact aaacgagctc tgcttatata
      121 ggtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc
      181 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
      241 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac
      301 cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata
      361 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
      421 gctcggtacc cgggtcgagg taggcgtgta cggtgggagg cctatataag cagagctcgt
      481 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      541 caccgggacc gatccagcct ccgcggcccc gaattcgagc tcggtacccg gggatcctct
      601 agtcagctga cgcgtatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      661 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      721 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
      781 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
      841 cccgaccaca tgaagcaaca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
      901 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
      961 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1021 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     1081 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     1141 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1201 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     1261 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1321 gagctgtaca agacgcgtaa aatggcgata cgggagctca aagtgtgcct tctcggggac
     1381 actggggttg ggaaatcaag catcgtgtgt cgatttgtcc aggatcactt tgaccacaac
     1441 atcagcccta ctattggggc atcttttatg accaaaactg tgccttgtgg aaatgaactt
     1501 cacaagttcc tcatctggga cactgctggt caggaacggt ttcattcatt ggctcccatg
     1561 tactatcgag gctcagctgc agctgttatc gtgtatgata ttaccaagca ggattcattt
     1621 tataccttga agaaatgggt caaggagctg aaagaacatg gtccagaaaa cattgtaatg
     1681 gccatcgctg gaaacaagtg cgacctctca gatattaggg aggttcccct gaaggatgct
     1741 aaggaatacg ctgaatctat aggtgccatc gtggttgaga caagtgcaaa aaatgctatt
     1801 aatatcgaag agctctttca aggaatcagc cgccagatcc cacccttgga cccccatgaa
     1861 aatggaaaca atggaacaat caaagttgag aagccaacca tgcaagccag ccgccggtgc
     1921 tgttgagcta gcgcggcctc gacgatatct ctagagctga gaacttcagg gtgagtttgg
     1981 ggacccttga ttgttctttc tttttcgcta ttgtaaaatt catgttatat ggagggggca
     2041 aagttttcag ggtgttgttt agaatgggaa gatgtccctt gtatcaccat ggaccctcat
     2101 gataattttg tttctttcac tttctactct gttgacaacc attgtctcct cttattttct
     2161 tttcattttc tgtaactttt tcgttaaact ttagcttgca tttgtaacga atttttaaat
     2221 tcacttttgt ttatttgtca gattgtaagt actttctcta atcacttttt tttcaaggca
     2281 atcagggtat attatattgt acttcagcac agttttagag aacaattgtt ataattaaat
     2341 gataaggtag aatatttctg catataaatt ctggctggcg tggaaatatt cttattggta
     2401 gaaacaacta caccctggtc atcatcctgc ctttctcttt atggttacaa tgatatacac
     2461 tgtttgagat gaggataaaa tactctgagt ccaaaccggg cccctctgct aaccatgttc
     2521 atgccttctt ctctttccta cagctcctgg gcaacgtgct ggttgttgtg ctgtctcatc
     2581 attttggcaa agaattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg
     2641 tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg
     2701 gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca
     2761 ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa
     2821 atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc
     2881 tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg
     2941 ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt
     3001 gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca
     3061 gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat
     3121 ccctcgactg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3181 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3241 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3301 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3361 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3421 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3481 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3541 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3601 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3661 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3721 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3781 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3841 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3901 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3961 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4021 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4081 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4141 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4201 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4261 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4321 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4381 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4441 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4501 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4561 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4621 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4681 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac
     4741 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4801 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4861 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4921 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4981 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5041 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5101 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     5161 gcgtatcacg aggccctttc gtcttcactc gacgcggcca gcttggtcga gctggatact
     5221 tcccgtccgc cagggggaca tgccggcgat gctgaaggtc gcgcgcattc ccgatgaaga
     5281 ggccggttac cgcctgttga cctggtggga cgggcagggc gccgcccgag tcttcgcctc
     5341 ggcggcgggc gctctgctca tggagcgcgc gtccggggcc ggggaccttg cacagatagc
     5401 gtggtccggc caggacgacg aggcttgcag gatcataatc agccatacca catttgtaga
     5461 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     5521 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5581 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5641 actcatcaat gtatcttatc atgtctggat cctctagaac taggtcgacg cggccg
//
