LOCUS       sequence-177910-        6175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A mammalian expression plasmid encoding CD8a without tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6175)
  AUTHORS   Madugula V, Lu L
  TITLE     A ternary complex comprising transportin1, Rab8 and the ciliary 
            targeting signal directs proteins to ciliary membranes.
  JOURNAL   J Cell Sci. 2016 Oct 15;129(20):3922-3934. Epub 2016 Sep 15.
  PUBMED    27633000
REFERENCE   2  (bases 1 to 6175)
  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 Cell 
            Sci."; date: "2016-10-15"; volume: "129(20)"; pages: "3922-3934. 
            Epub 2016 Sep 15"
COMMENT     Sequence Label: CD8a
FEATURES             Location/Qualifiers
     source          1..6175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        82..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          758..890
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     873..892
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        935..953
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      971..980
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1747..1868
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1758..1777)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1812..1831
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2054..2509
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2141..2160)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2351..2372
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2626..2983
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2626..2817
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2649..2669)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2834..2969
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3034..3828
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3088..3107)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3698..3717
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3892..3940
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3981..4000)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4100..4122
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4160..4178)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4246..4350
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4351..4419,4420..5211)
                     /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(4569..4588)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5382..5970
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5871..5890
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgttggcat tgattattga ctagttatta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg
      421 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      481 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactgcg atcgcccgcc
      541 ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcac tagaagcttt attgcggtag tttatcacag ttaaattgct
      661 aacgcagtca gtgcttctga cacaacagtc tcgaacttaa gctgcagtga ctctcttaag
      721 gtagccttgc agaagttggt cgtgaggcac tgggcaggta agtatcaagg ttacaagaca
      781 ggtttaagga gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg
      841 ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag gtgtccactc
      901 ccagttcaat tacagctctt aaggctagag tacttaatac gactcactat aggctagcct
      961 cgagaattca gccaccatgg ccttaccagt gaccgccttg ctcctgccgc tggccttgct
     1021 gctccacgcc gccaggccga gccagttccg ggtgtcgccg ctggatcgga cctggaacct
     1081 gggcgagaca gtggagctga agtgccaggt gctgctgtcc aacccgacgt cgggctgctc
     1141 gtggctcttc cagccgcgcg gcgccgccgc cagtcccacc ttcctcctat acctctccca
     1201 aaacaagccc aaggcggccg aggggctgga cacccagcgg ttctcgggca agaggttggg
     1261 ggacaccttc gtcctcaccc tgagcgactt ccgccgagag aacgagggct actatttctg
     1321 ctcggccctg agcaactcca tcatgtactt cagccacttc gtgccggtct tcctgccagc
     1381 gaagcccacc acgacgccag cgccgcgacc accaacaccg gcgcccacca tcgcgtcgca
     1441 gcccctgtcc ctgcgcccag aggcgtgccg gccagcggcg gggggcgcag tgcacacgag
     1501 ggggctggac ttcgcctgtg atatctacat ctgggcgccc ttggccggga cttgtggggt
     1561 ccttctcctg tcactggtta tcacccttta ctgcaaccac aggaaccgaa gacgtgtttg
     1621 caaatgtccc cggcctgtgg tcaaatcggg agacaagccc agcctttcgg cgagatacgt
     1681 ctaatctaga gtcgacccgg gcggccgctt ccctttagtg agggttaatg cttcgagcag
     1741 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     1801 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     1861 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     1921 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatccgataa ggatcgatcc
     1981 gggctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     2041 gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2101 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2161 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2221 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     2281 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2341 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     2401 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     2461 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt
     2521 attttctcct tacgcatctg tgcggtattt cacaccgcat acgcggatct gcgcagcacc
     2581 atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa
     2641 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     2701 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2761 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2821 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2881 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     2941 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttg attcttctga
     3001 cacaacagtc tcgaacttaa ggctagagcc accatgattg aacaagatgg attgcacgca
     3061 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3121 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3181 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     3241 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3301 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3361 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3421 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     3481 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     3541 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     3601 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     3661 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     3721 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     3781 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     3841 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgatggccg caataaaata
     3901 tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagc gataaggatc
     3961 cgcgtatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     4021 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4081 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4141 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4201 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4261 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4321 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4381 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4441 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4501 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4561 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4621 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4681 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4741 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4801 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4861 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4921 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4981 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5041 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5101 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5161 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5221 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5281 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5341 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5401 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5461 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5521 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5581 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5641 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5701 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5761 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5821 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5881 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5941 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6001 ctggcctttt gctggccttt tgctcacatg gctcgacaga tcttcaatat tggccattag
     6061 ccatattatt cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt
     6121 tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga ccgcc
//
