LOCUS       sequence-394409-        5695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of eGFP-Rab22 (human) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5695)
  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 5695)
  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-Rab22
FEATURES             Location/Qualifiers
     source          1..5695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1..3,4..6,7..717)
                     /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(46..67)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(307..326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             956..967
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1336..1470
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1360..1379)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1414..1433
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1833..1851)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1919..2023
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2024..2092,2093..2884)
                     /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     complement(2242..2261)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3055..3643
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3544..3563
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3797..3814
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4194..4213
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    4212..4267
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     4313..4332
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     intron          4464..5036
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     promoter        complement(5219..5257)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    join(5290..5308,5309..5331,5332..5350,5351..5373,5374..5392,
                     5393..5415,5416..5434,5435..5457,5458..5476,5477..5499,
                     5500..5518,5519..5541,5542..5560)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
      241 caacacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      481 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagttc
      721 tgcagcggcc gcatggcgct gagggagctc aaagtgtgtc tgctcgggga tacaggtgta
      781 ggtaaatcga gtattgtgtg gcggtttgtg gaagacagtt ttgatccaaa catcaaccca
      841 acaatagggg catcttttat gaccaagact gtccagtacc aaaatgagct acataaattc
      901 ctaatctggg atacagctgg acaagaacga tttcgtgcct tagcaccaat gtactatcga
      961 gggtcggctg cagctataat cgtttatgat atcacaaaag aagagacatt ttcaacatta
     1021 aagaattggg tgaaagagct tcgacagcat ggcccaccta atattgtagt tgccattgca
     1081 ggaaataaat gtgatcttat cgatgtaaga gaagtcatgg agagagatgc aaaggactac
     1141 gccgactcta ttcatgcaat ttttgtagag accagcgcaa aaaacgcgat aaacataaat
     1201 gaactcttta tagaaattag tcgaagaatt ccatccactg acgccaacct gccatctggc
     1261 ggtaagggct tcaaactccg aagacagcct tcagagccaa agcggagctg ctgctgagtc
     1321 gacctagttc tagaggatcc agacatgata agatacattg atgagtttgg acaaaccaca
     1381 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     1441 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     1501 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     1561 atggctgatt atgatcctgc aagcctcgtc gtcctggccg gaccacgcta tctgtgcaag
     1621 gtccccggcc ccggacgcgc gctccatgag cagagcgccc gccgccgagg cgaagactcg
     1681 ggcggcgccc tgcccgtccc accaggtcaa caggcggtaa ccggcctctt catcgggaat
     1741 gcgcgcgacc ttcagcatcg ccggcatgtc cccctggcgg acgggaagta tccagctcga
     1801 ccaagctggc cgcgtcgagt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     1861 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     1921 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     1981 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     2041 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     2101 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     2161 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     2221 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     2281 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     2341 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     2401 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     2461 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     2521 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     2581 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     2641 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     2701 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     2761 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     2821 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     2881 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     2941 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3001 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3061 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3121 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3181 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3241 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3301 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3361 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3421 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3481 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3541 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3601 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3661 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3721 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3781 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     3841 accgcctctc cccgcgcgtt ggccgattca ttaatgcagt cgagggatct ccataagaga
     3901 agagggacag ctatgactgg gagtagtcag gagaggagga aaaatctggc tagtaaaaca
     3961 tgtaaggaaa attttaggga tgttaaagaa aaaaataaca caaaacaaaa tataaaaaaa
     4021 atctaacctc aagtcaaggc ttttctatgg aataaggaat ggacagcagg gggctgtttc
     4081 atatactgat gacctcttta tagccaacct ttgttcatgg cagccagcat atgggcatat
     4141 gttgccaaac tctaaaccaa atactcattc tgatgtttta aatgatttgc cctcccatat
     4201 gtccttccga gtgagagaca caaaaaattc caacacacta ttgcaatgaa aataaatttc
     4261 ctttattagc cagaagtcag atgctcaagg ggcttcatga tgtccccata atttttggca
     4321 gagggaaaaa gatctcagtg gtatttgtga gccagggcat tggccacacc agccaccacc
     4381 ttctgatagg cagcctgcac ctgaggagtg aattctttgc caaaatgatg agacagcaca
     4441 acaaccagca cgttgcccag gagctgtagg aaagagaaga aggcatgaac atggttagca
     4501 gaggggcccg gtttggactc agagtatttt atcctcatct caaacagtgt atatcattgt
     4561 aaccataaag agaaaggcag gatgatgacc agggtgtagt tgtttctacc aataagaata
     4621 tttccacgcc agccagaatt tatatgcaga aatattctac cttatcattt aattataaca
     4681 attgttctct aaaactgtgc tgaagtacaa tataatatac cctgattgcc ttgaaaaaaa
     4741 agtgattaga gaaagtactt acaatctgac aaataaacaa aagtgaattt aaaaattcgt
     4801 tacaaatgca agctaaagtt taacgaaaaa gttacagaaa atgaaaagaa aataagagga
     4861 gacaatggtt gtcaacagag tagaaagtga aagaaacaaa attatcatga gggtccatgg
     4921 tgatacaagg gacatcttcc cattctaaac aacaccctga aaactttgcc ccctccatat
     4981 aacatgaatt ttacaatagc gaaaaagaaa gaacaatcaa gggtccccaa actcaccctg
     5041 aagttctcag ctctagagat atcgtcgagg ccgcgctagc acgcgtcagc tgactagagg
     5101 atccccgggt accgagctcg aattcggggc cgcggaggct ggatcggtcc cggtgtcttc
     5161 tatggaggtc aaaacagcgt ggatggcgtc tccaggcgat ctgacggttc actaaacgag
     5221 ctctgcttat ataggcctcc caccgtacac gcctacctcg acccgggtac cgagctcgac
     5281 tttcactttt ctctatcact gatagggagt ggtaaactcg actttcactt ttctctatca
     5341 ctgataggga gtggtaaact cgactttcac ttttctctat cactgatagg gagtggtaaa
     5401 ctcgactttc acttttctct atcactgata gggagtggta aactcgactt tcacttttct
     5461 ctatcactga tagggagtgg taaactcgac tttcactttt ctctatcact gatagggagt
     5521 ggtaaactcg actttcactt ttctctatca ctgataggga gtggtaaact cgacctatat
     5581 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
     5641 cctccataga agacaccggg accgatccag cctccgcggc cccgaattcc tgcag
//
