LOCUS       sequence-177921-        5540 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A mammalian expression plasmid encoding IL2R-alpha (interleukin-2 
            receptor subunit alpha) with C-terminus EGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5540)
  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 5540)
  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: IL2R-alpha-EGFP
FEATURES             Location/Qualifiers
     source          1..5540
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      15..603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     504..523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        778..1081
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1082..1285
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1236..1256
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2197..2206
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2203..2205,2206..2208,2209..2922)
                     /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(2203..2205,2206..2208,2209..2919)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2248..2269)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2509..2528)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2856..2877
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3045..3166
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3082..3101)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3136..3155
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3173..3628)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3310..3331)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3522..3541
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3655..3759
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3761..4118
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3761..3952
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3784..3804)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3969..4104
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4153..4947
                     /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             4153..4947
                     /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(4207..4226)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4817..4836
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5135..5154)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5179..5226
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
       61 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
      121 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
      181 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
      241 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
      301 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
      361 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
      421 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
      481 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
      541 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
      601 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
      661 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
      721 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      781 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      841 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      901 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      961 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     1021 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1081 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     1141 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     1201 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     1261 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg
     1321 gactcagatc tcgagatgga ttcatacctg ctgatgtggg gactgctcac gttcatcatg
     1381 gtgcctggct gccaggcaga gctctgtgac gatgacccgc cagagatccc acacgccaca
     1441 ttcaaagcca tggcctacaa ggaaggaacc atgttgaact gtgaatgcaa gagaggtttc
     1501 cgcagaataa aaagcgggtc actctatatg ctctgtacag gaaactctag ccactcgtcc
     1561 tgggacaacc aatgtcaatg cacaagctct gccactcgga acacaacgaa acaagtgaca
     1621 cctcaacctg aagaacagaa agaaaggaaa accacagaaa tgcaaagtcc aatgcagcca
     1681 gtggaccaag cgagccttcc aggtcactgc agggaacctc caccatggga aaatgaagcc
     1741 acagagagaa tttatcattt cgtggtgggg cagatggttt attatcagtg cgtccaggga
     1801 tacagggctc tacacagagg tcctgctgag agcgtctgca aaatgaccca cgggaagaca
     1861 aggtggaccc agccccagct catatgcaca ggtgaaatgg agaccagtca gtttccaggt
     1921 gaagagaagc ctcaggcaag ccccgaaggc cgtcctgaga gtgagacttc ctgcctcgtc
     1981 acaacaacag attttcaaat acagacagaa atggctgcaa ccatggagac gtccatattt
     2041 acaacagagt accaggtagc agtggccggc tgtgttttcc tgctgatcag cgtcctcctc
     2101 ctgagtgggc tcacctggca gcggagacag aggaagagta gaagaacaat ccgaattctg
     2161 cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca ccatggtgag caagggcgag
     2221 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     2281 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     2341 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     2401 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     2461 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     2521 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     2581 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     2641 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     2701 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     2761 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     2821 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     2881 gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg cgactctaga
     2941 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3001 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3061 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3121 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     3181 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3241 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3301 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3361 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3421 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3481 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3541 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3601 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3661 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3721 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     3781 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3841 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3901 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3961 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4021 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4081 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4141 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4201 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4261 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4321 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4381 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4441 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4501 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4561 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4621 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4681 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4741 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4801 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4861 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4921 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4981 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5041 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5101 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     5161 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     5221 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     5281 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5341 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5401 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5461 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5521 cgtcagaccc cgtagaaaag
//
