LOCUS       sequence-434311-        5959 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses N-terminally EGFP-tagged IFT38 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5959)
  AUTHORS   Katoh Y, Terada M, Nishijima Y, Takei R, Nozaki S, Hamada H, 
            Nakayama K
  TITLE     Overall architecture of the intraflagellar transport (IFT)-B complex
            containing Cluap1/IFT38 as an essential component of the IFT-B 
            peripheral subcomplex.
  JOURNAL   J Biol Chem. 2016 Mar 15. pii: jbc.M116.713883.
  PUBMED    26980730
REFERENCE   2  (bases 1 to 5959)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem. 2016 Mar 15. pii: jbc.M116.713883."
COMMENT     Sequence Label: pEGFP-C1-IFT38
FEATURES             Location/Qualifiers
     source          1..5959
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2747..2868
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2784..2803)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2838..2857
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2875..3330)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3012..3033)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3224..3243
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3357..3461
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3463..3820
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3463..3654
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3486..3506)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3671..3806
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3855..4649
                     /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             3855..4649
                     /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(3909..3928)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4519..4538
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4837..4856)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4881..4928
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5257..5845
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5746..5765
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctatgtct ttccgcgacc tccgcaattt cacagagatg
     1381 atgagagccc tgggataccc tcgacatatt tctatggaaa atttccgtac acccaatttt
     1441 ggacttgtat ctgaagtgct tctctggctt gtgaaaagat atgagcccca gactgacatc
     1501 ccgcctgacg tggatactga acaggaccga gttttcttca ttaaggcaat tgcccagttc
     1561 atggccacca aggcacatat aaaactcaac actaagaagc tttatcaagc agatgggtat
     1621 gcggtaaaag agctgctgaa gatcacatct gtcctttata atgctatgaa gaccaagggg
     1681 atggagggct ctgaaatagt agaggaagat gtcaacaagt tcaagtttga tcttggctca
     1741 aagattgcag atttgaaggc agccaggcag cttgcgtctg aaatcacctc caaaggagca
     1801 tctctgtatg acttgctcgg catggaagta gagttgaggg aaatgagaac agaagccatt
     1861 gccagacctc tggaaataaa cgagactgaa aaagtgatga gaattgcaat aaaagagatt
     1921 ttgacacagg ttcagaagac taaagacctg ctcaataatg tggcctctga tgaagctaat
     1981 ttagaagcca aaatcgaaaa gagaaaatta gaactggaaa gaaatcggaa gcgactagag
     2041 actctgcaga gtgtcaggcc atgttttatg gatgagtatg agaagactga ggaagaatta
     2101 caaaagcagt atgacactta tctggagaaa tttcaaaatc tgacttatct ggaacaacag
     2161 cttgaagacc atcataggat ggagcaagaa aggtttgagg aagctaaaaa cactctctgc
     2221 ctgatacaga acaagctcaa ggaggaagag aagcgcctgc tcaagagtgg aagtaacgat
     2281 gactcggaca tagacatcca ggaggacgat gaatccgaca gtgagttgga agaaaggcgg
     2341 ctgcccaagc cacagacagc catggagatg ctcatgcaag gaagacctgg caaacgcatt
     2401 gtgggcacga tgcaaggtgg agactccgat gacaatgagg actcggagga gagtgaaatt
     2461 gacatggaag atgatgatga cgaggatgac gatttggaag acgagagcat ttctctctca
     2521 ccaaccaagc ccaatcgaag ggtccggaaa tctgaacccc tggatgagag tgacaatgac
     2581 ttctgagaat tctgcagtcg acggtaccgc gggcccggga tccaccggat ctagataact
     2641 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     2701 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     2761 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     2821 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg
     2881 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     2941 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     3001 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3061 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3121 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3181 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3241 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3301 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     3361 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3421 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     3481 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3541 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3601 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3661 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3721 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     3781 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     3841 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     3901 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     3961 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4021 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4081 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4141 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4201 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4261 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4321 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4381 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4441 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4501 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4561 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4621 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     4681 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     4741 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     4801 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     4861 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     4921 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     4981 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     5041 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     5101 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5161 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5221 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5281 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5341 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5401 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5461 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5521 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5581 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5641 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5701 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5761 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5821 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5881 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     5941 ccgtattacc gccatgcat
//
