LOCUS       sequence-493800-        5791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mammalian Arl13b with RVEP4A mutation and GFP-tagged.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5791)
  AUTHORS   Mahajan D, Madugula V, Lu L
  TITLE     Rab8 and TNPO1 are ciliary transport adaptors for GTPase Arl13b by 
            interacting with its RVEP motif containing ciliary targeting 
            sequence.
  JOURNAL   J Biol Chem. 2023 May;299(5):104604. doi: 10.1016/j.jbc.2023.104604.
            Epub 2023 Mar 11.
  PUBMED    36907439
REFERENCE   2  (bases 1 to 5791)
  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: "2023-05"; volume: "299(5):104604. doi"; pages: " 
            10.1016/j.jbc.2023.104604. Epub 2023 Mar 11"
COMMENT     Sequence Label: Arl13b-RVEP-4A-GFP
FEATURES             Location/Qualifiers
     source          1..5791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        172..375
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     326..346
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1731..1740
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1737..1739,1740..1742,1743..2456)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1737..1739,1740..1742,1743..2453)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1782..1803)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2043..2062)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2390..2411
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2579..2700
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2616..2635)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2670..2689
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2707..3162)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2844..2865)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3056..3075
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3189..3293
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3295..3652
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3295..3486
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3318..3338)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3503..3638
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3687..4481
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3687..4481
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3741..3760)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4351..4370
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4669..4688)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4713..4760
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5089..5677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5578..5597
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
      121 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      181 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      241 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      301 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      361 tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg gactcagatc
      421 tcgaggccac catgttcagt ctgatggcca gttgctgcgg ctggttcaag cggtggcggg
      481 agcctgtcag aaaggtgact cttttgatgg tgggacttga taatgctggt aaaaccgcaa
      541 cagcaaaggg aatccaagga gaataccctg aagatgtagc tcctactgtt ggattttcaa
      601 aaattaacct tagacaagga aagtttgaag tcaccatctt tgacttggga ggtggaataa
      661 gaattcgggg aatctggaag aattactatg ctgaatccta tggggtaata tttgttgtgg
      721 attccagtga tgaagagaga atggaagaga caaaagaggc tatgtcagaa atgctaagac
      781 atcctaggat atcgggaaag cctatattgg tgttggcaaa taaacaagat aaagaaggag
      841 ctttaggaga agctgatgtc attgaatgtc tatctctgga aaaattggtc aatgagcaca
      901 agtgcctgtg tcagatagaa ccatgttcag caatctcggg gtatggaaag aaaattgaca
      961 agtccattaa aaaaggcctt tattggctgc tacatgttat tgcaagagac tttgatgcct
     1021 taaatgaacg catccaaaaa gagacaacag agcagcgtgc tcttgaggaa caagagaaac
     1081 aagaaagagc tgaacgagtg cgaaaattac gagaagaaag aaaacaaaat gaacaggagc
     1141 aggctgaact cgatggaacc agtggtctgg ctgagttgga cccagaacca acgaatcctt
     1201 tccagccaat agcatctgta atcattgaga acgaaggaaa acttgaaaga gagaaaaaaa
     1261 accaaaaaat ggagaaagac agtgatggct gccacctgaa acataaaatg gagcatgagc
     1321 aaatagagac acaaggccag gttaatcaca atggccaaaa aaataatgaa tttggactag
     1381 tagaaaatta taaggaggca ttaacacagc agttaaagaa tgaagatgag acagaccggc
     1441 catcattgga atcagctaat ggtaaaaaga aaactaagaa actaagaatg aaaaggaacc
     1501 acgcggcagc agcacttaat atagatgact gtgctcctga gagtccaacg ccacccccac
     1561 cccctcctcc tgttggctgg ggaaccccta aagtcactag acttccaaaa cttgagcctc
     1621 ttggtgaaac acatcataat gatttctata ggaagccact gcctcccctg gctgtgccac
     1681 agcgacctaa cagtgatgct catgatgtga tctcattgga tccaccggtc gccaccatgg
     1741 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1801 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1861 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1921 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     1981 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     2041 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     2101 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     2161 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     2221 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     2281 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     2341 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2401 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg
     2461 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     2521 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     2581 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     2641 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     2701 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2761 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     2821 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2881 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2941 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3001 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3061 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     3121 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     3181 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3241 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     3301 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3361 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3421 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3481 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3541 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3601 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     3661 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3721 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3781 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3841 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     3901 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3961 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4021 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4081 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4141 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4201 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4261 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4321 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4381 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4441 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4501 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4561 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4621 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     4681 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     4741 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     4801 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     4861 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     4921 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4981 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5041 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5101 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5161 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5221 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5281 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5341 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5401 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5461 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5521 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5581 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5641 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5701 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5761 ttctgtggat aaccgtatta ccgccatgca t
//
