LOCUS       sequence-494683-        5339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CD8a fused with a segment of the Arl13b protein (amino 
            acids 354-363) and GFP-tagged.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5339)
  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 5339)
  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: CD8a-AA354-363-GFP
FEATURES             Location/Qualifiers
     source          1..5339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1279..1288
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1285..1287,1288..1290,1291..2004)
                     /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(1285..1287,1288..1290,1291..2001)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1330..1351)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1591..1610)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1938..1959
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2127..2248
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2164..2183)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2218..2237
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2255..2710)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2392..2413)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2604..2623
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2737..2841
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2843..3200
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2843..3034
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2866..2886)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3051..3186
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3235..4029
                     /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             3235..4029
                     /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(3289..3308)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3899..3918
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4217..4236)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4261..4308
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4637..5225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5126..5145
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagat ggccttacca gtgaccgcct tgctcctgcc gctggccttg
      661 ctgctccacg ccgccaggcc gagccagttc cgggtgtcgc cgctggatcg gacctggaac
      721 ctgggcgaga cagtggagct gaagtgccag gtgctgctgt ccaacccgac gtcgggctgc
      781 tcgtggctct tccagccgcg cggcgccgcc gccagtccca ccttcctcct atacctctcc
      841 caaaacaagc ccaaggcggc cgaggggctg gacacccagc ggttctcggg caagaggttg
      901 ggggacacct tcgtcctcac cctgagcgac ttccgccgag agaacgaggg ctactatttc
      961 tgctcggccc tgagcaactc catcatgtac ttcagccact tcgtgccggt cttcctgcca
     1021 gcgaagccca ccacgacgcc agcgccgcga ccaccaacac cggcgcccac catcgcgtcg
     1081 cagcccctgt ccctgcgccc agaggcgtgc cggccagcgg cggggggcgc agtgcacacg
     1141 agggggctgg acttcgcctg tgatatctac atctgggcgc ccttggccgg gacttgtggg
     1201 gtccttctcc tgtcactggt tatcaccgaa ttcaaaagga accaccgggt agaaccactt
     1261 aatcaggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
     1321 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1381 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     1441 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     1501 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1561 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1621 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1681 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     1741 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     1801 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1861 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     1921 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1981 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
     2041 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2101 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     2161 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2221 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     2281 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2341 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     2401 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     2461 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     2521 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     2581 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     2641 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     2701 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2761 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2821 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     2881 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2941 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3001 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3061 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3121 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3181 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3241 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3301 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3361 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     3421 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3481 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3541 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3601 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3661 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3721 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     3781 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3841 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3901 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     3961 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4021 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4081 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4141 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4201 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     4261 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     4321 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     4381 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     4441 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     4501 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4561 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4621 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4681 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4741 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4801 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4861 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4921 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4981 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5041 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5101 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5161 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5221 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5281 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
