LOCUS       sequence-177941-        5906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A mammalian expression plasmid encoding fibrocystin mimentic clone 
            CFF (CD8a luminal domain, fibrocystin transmembrane and cytosolic 
            domains) with C-terminus EGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5906)
  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 5906)
  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: CFF-EGFP
FEATURES             Location/Qualifiers
     source          1..5906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      3..591
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     492..511
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        766..1069
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1070..1273
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1224..1244
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2551..2560
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2557..2559,2560..2562,2563..3276)
                     /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(2557..2559,2560..2562,2563..3273)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2602..2623)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2863..2882)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3210..3231
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3399..3520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3436..3455)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3490..3509
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3527..3982)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3664..3685)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3876..3895
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4009..4113
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4115..4472
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4115..4306
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4138..4158)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4323..4458
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4507..5301
                     /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             4507..5301
                     /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(4561..4580)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5171..5190
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5489..5508)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5533..5580
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
       61 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
      121 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
      181 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      241 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      301 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      361 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
      421 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      481 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      541 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      601 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      661 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
      721 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
      781 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      841 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      901 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      961 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     1021 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     1081 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     1141 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     1201 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     1261 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga ctcagatctc
     1321 gagatggcct taccagtgac cgccttgctc ctgccgctgg ccttgctgct ccacgccgcc
     1381 aggccgagcc agttccgggt gtcgccgctg gatcggacct ggaacctggg cgagacagtg
     1441 gagctgaagt gccaggtgct gctgtccaac ccgacgtcgg gctgctcgtg gctcttccag
     1501 ccgcgcggcg ccgccgccag tcccaccttc ctcctatacc tctcccaaaa caagcccaag
     1561 gcggccgagg ggctggacac ccagcggttc tcgggcaaga ggttggggga caccttcgtc
     1621 ctcaccctga gcgacttccg ccgagagaac gagggctact atttctgctc ggccctgagc
     1681 aactccatca tgtacttcag ccacttcgtg ccggtcttcc tgccagcgaa gcccaccacg
     1741 acgccagcgc cgcgaccacc aacaccggcg cccaccatcg cgtcgcagcc cctgtccctg
     1801 cgcccagagg cgtgccggcc agcggcgggg ggcgcagtgc acacgagggg gctggacttc
     1861 gcctgtgatg agagatcaac tatcatcttg gccttgtccc tgtgctcagt ggcatcatgg
     1921 gtggctctga gctgtctcgt ttgctgctgg tttaagaaaa gcaaaaccag aaaaataaaa
     1981 ccagaagaca tatctgaatc ccaggctaag gaacaaaaga agaataccca taattcctcc
     2041 aaacccagag gactacaagc aaagacagca aaagagaaca ctttgatggg agaagatatg
     2101 agaatgaagg tcatgcaggg aatgcagagc cagtttcccc aacactcaat ggatggagtg
     2161 tccaaaagga aagttagccg ccttgctgtc acaggggaaa gaacagctac acctgcccca
     2221 aagattccca gaatcacctg tgttccagga tctcttgctc agcagctgac actgcaggag
     2281 cctgggaact ggcaggaggc ccaacagcag ttgctcagat accagctggc aggccgcaat
     2341 cagctgctcc tgttacgccc agacctcagg caagagagga aacagggtca ggagcctagc
     2401 cagctggaca aagggagtga ctgcactgga ctgtctcaag agaaggccac ttgcattccc
     2461 actgagactt tcggcctcca cactactcca ccagaaacca tccagcgaat tctgcagtcg
     2521 acggtaccgc gggcccggga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg
     2581 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     2641 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     2701 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     2761 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     2821 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     2881 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     2941 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     3001 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     3061 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     3121 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     3181 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     3241 gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa
     3301 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     3361 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3421 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3481 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     3541 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3601 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3661 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3721 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3781 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3841 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3901 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3961 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4021 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4081 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     4141 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4201 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4261 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4321 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4381 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     4441 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     4501 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4561 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4621 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4681 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4741 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4801 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4861 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4921 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4981 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     5041 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5101 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5161 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5221 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5281 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5341 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5401 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5461 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     5521 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     5581 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     5641 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     5701 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     5761 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5821 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5881 accccgtaga aaagatcaaa ggatct
//
