LOCUS       sequence-408797-        6217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 3xFlag-tagged mouse CEP83 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6217)
  TITLE     Mammalian expression plasmids for ciliary genes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6217)
  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: 
            "Unpublished"
COMMENT     Sequence Label: 3xFlag-CEP83
FEATURES             Location/Qualifiers
     source          1..6217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             81..146
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2269..2403
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2293..2312)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2347..2366
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2516..2536)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2516..2534)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2555..2571)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2555..2571)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2568..2590)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2579..2595
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2603..2609,2610..2627,2628..2633))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2648..2669
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2786..2803)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2957..3545)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3037..3056)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3716..4507,4508..4576))
                     /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     4339..4358
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4577..4681)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4707..5162)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4844..4865)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5056..5075
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5233..6217
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agacttgttc
       61 tttttgcagg atctaccatg gactacaaag accatgacgg tgattataaa gatcatgaca
      121 tcgattacaa ggatgacgat gacaagggat ccatggacac atttcctagc ctatttccac
      181 ctggaggaga tagtaggctg aatcctgagc ctgagttcca aaatatgtta attgatgaaa
      241 gggtacgctg tgaacatcat aaacataatt atcaggctct gaaaattgaa cacaaaaggt
      301 tgcaggaaga atatgtaaaa tcacaaaatg aacttaaacg tgtattaatt gaaaagcaag
      361 caagccagga aaaattccaa ctgctccttg aagacttaag gggagaatta gtagagaaag
      421 ctagagacat agaaaaaatg aaactgcagg tactaacacc acaaaaattg gaattggtaa
      481 aagcccaact acaacaagaa ttagaagctc caatgcgaga acgttttcgg actcttgatg
      541 aagaagtgga aaggtacaga gctgagtata acaagctgcg ctacgagtat acatttctca
      601 agtcagagtt tgaacaccag aaagaagagt ttactcgggt ttcagaagaa gagaaaatga
      661 aatacaagtc agaggttgca cgactggaga aggacaaaga ggagctacat aaccagctgc
      721 ttagtgttga tcccacgaga gacagcaaac gaatggagca acttgttcga gaaaaaaccc
      781 atttgcttca gaaattgaaa agtttagagg ctgaagtagc agaattaagg gctgagaaag
      841 aaaattctgg tgctcaggta gaaaatgtcc aaagaataca ggtgaggcag ttggctgaga
      901 tgcaggctac actcagatcc ttggaggctg aaaagcagtc agctaaacta caagctgagc
      961 gtttagaaaa agaactacaa tcaagcaatg aacagaatac ctgcttaatc agcaaactgc
     1021 atagagctga ccgagaaatc agcacactgg ccagtgaagt gaaagagctt aaacatgcaa
     1081 acaaactaga aataactgac atcaaactgg aggcagcaag agctaagagt gagctcgaaa
     1141 gagaaaggaa taagatccaa agtgaacttg atggattgca gtcagacaat gaaattctca
     1201 agtcaactgt tgaacaccac aaagcgctct tagtggaaaa agatcgtgaa ttaatacgta
     1261 aagtacaagc tgccaaggaa gaaggttacc agaaactcat ggtcttacaa gatgaaaagc
     1321 tagaacttga gaacagatta tcagacttag agaagatgaa ggtggagcgt gatgtctgga
     1381 ggcagtcgga aaaggagcag tgtgaggaga agctgcgggc ctcccagatg gcagaggagg
     1441 cagcccgcag ggagctgcag agcactaggt taaaacttca acaacaaatt gtgaataccg
     1501 aaaaagcaga aaaagaaaaa cttgaaaatt ctgaactgaa acagcaaatc agtcatttgc
     1561 agatccaagt gacctcactg acacagtcgg agaacgactt actgaattca aaccacatgc
     1621 tgaaggacat ggtggagaga ctaaaacaag aatgccgaaa tttaaggagc caagctgaga
     1681 aagcacaact agacgttgaa aagacactgg aggaaaaaca gatccagtgg ttggaagaaa
     1741 agcacaagct ccatgagcga atcacagaca gagaagaaaa gtataaccaa gccaaggaaa
     1801 agctacagcg cgcagcgact gctcagaaaa agagaaaatc tcttcatgag aacaaattga
     1861 agagactaca ggagaaagta gaagtcttgg aggcaaagaa agaagaacta gaaacagaaa
     1921 accaggtgct aaatagacaa aatgttccat ttgaagaata tacaaggctt caaaaacgac
     1981 taaaggacat acagagaaga cataatgaat ttcgaagtct aattttggtt cctaacatgc
     2041 ctccaacagc atcgattagt cctgctaact tccagtccgc agtgacagtt cctggagcgg
     2101 aactctcttt tccccctcat ctgcaggagg aacaacatca aagagagctc tctctccttc
     2161 gcaaaagact ggaagaactg gagactacac aaagaaaaca actggaggag cttgggtctc
     2221 ctggggagtg actcgagcct ctagaactat agtgagtcgt attacgtaga tccagacatg
     2281 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     2341 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     2401 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     2461 ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt
     2521 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     2581 gttatccgct cacaattcca cacaacatac gagccgggag cataaagtgt aaagcctggg
     2641 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     2701 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2761 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2821 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2881 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2941 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3001 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3061 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3121 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3181 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3241 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3301 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3361 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3421 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3481 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3541 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3601 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3661 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3721 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3781 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3841 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3901 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3961 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4021 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4081 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4141 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4201 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4261 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4321 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4381 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4441 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4501 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4561 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4621 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4681 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     4741 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     4801 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     4861 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     4921 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     4981 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     5041 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     5101 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     5161 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     5221 gctattacgc cagtcgaccg ccaattcaat atggcgtata tggactcatg ccaattcaat
     5281 atggtggatc tggacctgtg ccaattcaat atggcgtata tggactcgtg ccaattcaat
     5341 atggtggatc tggaccccag ccaattcaat atggcggact tggcaccatg ccaattcaat
     5401 atggcggacc tggcactgtg ccaactgggg aggggtctac ttggcacggt gccaagtttg
     5461 aggaggggtc ttggccctgt gccaagtccg ccatattgaa ttggcatggt gccaataatg
     5521 gcggccatat tggctatatg ccaggatcaa tatataggca atatccaata tggccctatg
     5581 ccaatatggc tattggccag gttcaatact atgtattggc cctatgccat atagtattcc
     5641 atatatgggt tttcctattg acgtagatag cccctcccaa tgggcggtcc catataccat
     5701 atatggggct tcctaatacc gcccatagcc actcccccat tgacgtcaat ggtctctata
     5761 tatggtcttt cctattgacg tcatatgggc ggtcctattg acgtatatgg cgcctccccc
     5821 attgacgtca attacggtaa atggcccgcc tggctcaatg cccattgacg tcaataggac
     5881 cacccaccat tgacgtcaat gggatggctc attgcccatt catatccgtt ctcacgcccc
     5941 ctattgacgt caatgacggt aaatggccca cttggcagta catcaatatc tattaatagt
     6001 aacttggcaa gtacattact attggaagta cgccagggta cattggcagt actcccattg
     6061 acgtcaatgg cggtaaatgg cccgcgatgg ctgccaagta catccccatt gacgtcaatg
     6121 gggaggggca atgacgcaaa tgggcgttcc attgacgtaa atgggcggta ggcgtgccta
     6181 atgggaggtc tatataagca atgctcgttt agggaac
//
