LOCUS       sequence-266682-        9109 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9109)
  AUTHORS   Graser S, Stierhof YD, Lavoie SB, Gassner OS, Lamla S, Le Clech M, 
            Nigg EA
  TITLE     Cep164, a novel centriole appendage protein required for primary 
            cilium formation.
  JOURNAL   J Cell Biol. 2007 Oct 22;179(2):321-30.
  PUBMED    17954613
REFERENCE   2  (bases 1 to 9109)
  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 
            Biol."; date: "2007-10-22"; volume: "179(2)"; pages: "321-30"
COMMENT     Sequence Label: pEGFP Cep164 (Nigg CW324)
FEATURES             Location/Qualifiers
     source          1..9109
                     /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      5055..5064
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5061..5063,5064..5066,5067..5780)
                     /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(5061..5063,5064..5066,5067..5777)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5106..5127)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5367..5386)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5714..5735
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5897..6018
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5934..5953)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5988..6007
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6025..6480)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6162..6183)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6374..6393
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6507..6611
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6613..6970
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6613..6804
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6636..6656)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6821..6956
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             7005..7799
                     /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             7005..7799
                     /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(7059..7078)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7669..7688
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7987..8006)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8031..8078
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8407..8995
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8896..8915
                     /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 atctcgagct caagcttcga attcggccgg cctaaccatg gctggacgac
      661 ccctccgcat aggagatcag ctggttctgg aagaagatta tgatgagacc tacattccta
      721 gtgagcaaga aattcttgaa tttgcccggg agattggtat tgatcccatc aaggaaccag
      781 aactgatgtg gctggcgcga gagggcatcg tggccccact gcctggagag tggaaaccat
      841 gccaggacat cacaggtgac atttactatt tcaacttcgc caacgggcag tctatgtggg
      901 accatccatg tgacgaacac tatcggagct tggtgatcca agagcgggca aagctgtcaa
      961 cttctggggc cattaagaag aagaaaaaaa aaaaggaaaa gaaagacaag aaggacagag
     1021 acccccccaa aagttcgctg gccttgggtt cctcattagc cccagttcat gttcctcttg
     1081 ggggcctggc tcctttacga ggtcttgtgg ataccccacc ctctgctctt cgtggatctc
     1141 aaagcgtgag cctggggagc tcagtggagt ctggacgtca gcttggagaa ctcatgctgc
     1201 cttcacaggg tctcaagacc tctgcttata caaagggtct cttgggctcc atatatgagg
     1261 acaagactgc tctcagcctc ttgggtttag gagaagaaac caatgaggag gatgaggagg
     1321 aaagtgacaa ccagagtgtc cacagctcaa gtgagcctct taggaaccta cacctggaca
     1381 ttggggcact ggggggtgac tttgagtatg aggagtctct gagaacaagc cagccagagg
     1441 agaagaagga tgtttctctg gattcagatg ctgccggtcc ccctactccc tgcaagccct
     1501 ccagcccagg tgcagacagc agtctgagca gtgctgttgg caaagggcga cagggaagtg
     1561 gagcaagacc tggtcttcca gaaaaagagg aaaatgagaa gagtgaacct aagatttgca
     1621 ggaatctggt gacccccaag gcagacccta caggcagtga gcctgccaaa gcctctgaaa
     1681 aggaagcacc agaggacaca gtagatgcag gagaggaggg ttccaggagg gaagaggcag
     1741 ccaaggagcc aaagaagaag gcttctgctc tggaagaggg cagttcagac gccagccaag
     1801 aactggaaat tagtgaacac atgaaggaac cacagctctc agactccata gcttctgacc
     1861 ccaagtcctt ccatggcctg gacttcggtt ttcgcagccg gatctcggag cacctgctgg
     1921 atgttgatgt gctttcccca gtcctgggtg gagcttgtcg gcaggcccag caaccactgg
     1981 gaatagaaga caaggatgac agccagtcca gccaagatga gctgcagagc aagcagtcca
     2041 aaggcctgga ggagaggtac cataggttat ctcctccact tccacacgag gagcgggccc
     2101 agagtccccc tcgcagcctg gccactgaag aagagccccc ccagggcccc gaggggcagc
     2161 ccgagtggaa ggaggcagag gagcttgggg aggactctgc agccagcctc agcctgcagc
     2221 tgtccctcca gagggagcag gccccaagcc cacctgctgc ctgtgagaag ggcaaggagc
     2281 agcattccca ggccgaggag ctgggccctg ggcaggaaga ggcagaggat cctgaggaga
     2341 aggtggcggt cagccccacc ccgccagtct ctccagaggt gcgatccaca gagcctgtgg
     2401 ctcccccaga gcagctctca gaggctgcac taaaggccat ggaagaggca gtggcccaag
     2461 tactcgagca agaccagagg cacctgctgg aatccaagca agagaagatg cagcaactgc
     2521 gggagaagct gtgccaagag gaggaagagg agatcctccg gcttcaccag cagaaagagc
     2581 aatctctcag ttccttgagg gagcggctgc agaaagccat tgaggaggag gaggcccgga
     2641 tgagagagga ggaaagccag aggctatcct ggctccgagc tcaggtccag tccagcacac
     2701 aagcagatga ggaccaaatc agggctgagc aagaggcttc cctgcagaaa ctgagagaag
     2761 agttggagtc tcaacagaag gctgagaggg ccagcttgga acagaaaaat aggcaaatgc
     2821 tggagcagct caaggaagag atagaggctt cggagaagag cgagcaggct gccctgaatg
     2881 ctgcaaagga gaaggctctg cagcagctga gggagcagct ggaaggggag aggaaagaag
     2941 ctgtggcaac gctggagaag gagcacagtg ctgagctgga gcggctctgc tcctcattgg
     3001 aggccaagca ccgggaggtg gtctccagcc tccagaagaa gatacaggaa gctcaacaga
     3061 aagaggaggc ccagctgcag aagtgccttg ggcaagtgga gcacagagtt caccagaagt
     3121 cttatcacgt ggctgggtat gagcacgagc tcagcagtct cctgcgagag aagcgccagg
     3181 aagtggaagg ggagcatgag aggaggttgg acaagatgaa ggaggagcac cagcaagtga
     3241 tggctaaggc cagagagcag tatgaagctg aggagaggaa gcagcgggct gagcttctgg
     3301 ggcacctgac cggagagctg gagcgcctgc agagggccca tgaacgagaa ctggagactg
     3361 tgaggcagga gcaacacaag cgtcttgagg acttgcggcg ccggcacagg gagcaggaaa
     3421 ggaagctcca ggatttagag ttggaccttg aaaccagagc taaagatgtc aaggccagat
     3481 tggctctgct ggaggtccag gaggagaccg cccggaggga gaagcagcag ctgcttgatg
     3541 tgcagaggca ggttgctctg aagagtgagg aagccacagc cacccatcag cagctggagg
     3601 aggcacagaa ggagcacacc cacctgttgc agtcaaacca gcagctccga gaaattcttg
     3661 atgagctgca ggcccgcaag ctgaagctgg agtcccaagt ggatctgctg caggctcaga
     3721 gccagcaact gcagaaacac ttcagcagcc tggaggctga agctcaaaag aagcagcacc
     3781 tgttgagaga agtgacagtt gaggaaaata atgcttcccc acattttgag ccagatctcc
     3841 atattgagga cctgaggaaa tcccttggaa caaaccagac caaagaggtg tcttcttctc
     3901 tctcccagag caaggaggac ttatacttgg acagcctgtc ctcccacaat gtctggcacc
     3961 tcctctctgc tgagggggta gccctccgta gtgccaagga gttccttgtg cagcagacac
     4021 gctccatgcg gaggcggcag acagctctga aagctgccca gcagcattgg cgccatgagc
     4081 tggccagtgc gcaggaggtg gccaaagacc caccaggcat caaggccctg gaagatatgc
     4141 gcaagaacct ggagaaggag accaggcacc tggatgagat gaagtcggcc atgcggaaag
     4201 gccacaacct gctgaagaag aaagaggaga agctgaatca gttggagtcc tctctttggg
     4261 aagaggcctc agatgagggc actctgggag gatcccccac caagaaggca gtaaccttcg
     4321 acctcagtga catggacagc ctgagcagtg aaagttctga atctttttcc ccgcctcacc
     4381 tcgactcaac cccgagtctc acctcccgca agatccacgg gcttagccac tccctccggc
     4441 agatcagcag ccagctgagc agtgtcctca gcatcctgga cagcctcaac cctcagtcgc
     4501 cgccgccgct cctcgcctcc atgccagccc agctccctcc ccgggaccct aagagcaccc
     4561 ccacccccac ctactatggc tccctggcca ggttctcagc cttatcatct gctacaccca
     4621 cgtccaccca atgggcctgg gattcagggc aggggcccag gctcccctcc tctgtggctc
     4681 aaacggtgga cgacttcctg ttggagaagt ggcgcaagta ttttccatct ggcatcccgc
     4741 tgctcagcaa cagccccacc ccgctggaga gcaggctggg ttacatgtct gccagtgagc
     4801 agctccggct cctacagcac tcccattcgc aagtccctga ggcgggcagc accacctttc
     4861 agggcataat tgaggccaac cggaggtggc tggaacgtgt caagaatgac cccaggttac
     4921 ctctcttctc ttcaacaccc aagccaaaag ctactttgag cctcctgcag ctgggccttg
     4981 atgagcacaa cagagtgaag gtgtatcgct tcgcggccgc tgcagtcgac ggtaccgcgg
     5041 gcccgggatc catcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     5101 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     5161 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     5221 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     5281 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     5341 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     5401 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     5461 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     5521 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     5581 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     5641 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     5701 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     5761 tggacgagct gtacaagtaa agcggcccta gatcataatc agccatacca catttgtaga
     5821 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     5881 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5941 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6001 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     6061 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     6121 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     6181 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     6241 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     6301 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     6361 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     6421 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     6481 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     6541 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6601 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     6661 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     6721 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6781 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     6841 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     6901 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     6961 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     7021 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     7081 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     7141 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     7201 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     7261 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     7321 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     7381 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     7441 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     7501 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     7561 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     7621 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     7681 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     7741 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     7801 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     7861 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     7921 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     7981 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     8041 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     8101 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     8161 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     8221 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     8281 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     8341 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     8401 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     8461 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     8521 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     8581 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     8641 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     8701 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     8761 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     8821 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     8881 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     8941 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     9001 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     9061 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
