LOCUS       sequence-252631-        7132 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Smoothened.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7132)
  AUTHORS   Ott C, Elia N, Jeong SY, Insinna C, Sengupta P, Lippincott-Schwartz 
            J
  TITLE     Primary cilia utilize glycoprotein-dependent adhesion mechanisms to 
            stabilize long-lasting cilia-cilia contacts.
  JOURNAL   Cilia. 2012 Apr 25;1(1):3. doi: 10.1186/2046-2530-1-3.
  PUBMED    23351752
REFERENCE   2  (bases 1 to 7132)
  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: "Cilia."; 
            date: "2012-04-25"; volume: "1(1):3. doi"; pages: " 
            10.1186/2046-2530-1-3"
COMMENT     Sequence Label: SmoCFP
FEATURES             Location/Qualifiers
     source          1..7132
                     /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      3008..3017
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3014..3016,3017..3019,3020..3730)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3059..3080)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3320..3339)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3667..3688
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3731..3796
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3920..4041
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3957..3976)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4011..4030
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4048..4503)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4185..4206)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4397..4416
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4530..4634
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4636..4993
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4636..4827
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4659..4679)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4844..4979
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5028..5822
                     /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             5028..5822
                     /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(5082..5101)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5692..5711
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6010..6029)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6054..6101
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6430..7018
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6919..6938
                     /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 gctacaagct
      601 taccatggcc gctggccgcc ccgtgcgtgg gcccgagctg gcgccccgga ggctgctgca
      661 gttgctgctg ctggtactgc tggggggccc gggccggggg gcggccttga gcgggaacgt
      721 gaccgggcct gggcctcaca gcgccagcgg gagctcgagg aggaacgtgc cggtgaccag
      781 ccctccgccg ccgctgctga gccactgcgg ccgggccgcc cactgcgagc ctttgcgcta
      841 caacgtgtgc ctgggctcgg cgctgcccta cggagccacc accacgctgc tggctgggga
      901 ctcggactcg caggaggaag cgcacggcaa gctcgtgctc tggtccggcc tccggaatgc
      961 cccccgctgc tgggcagtga tccagcccct gctgtgtgct gtctacatgc ccaagtgtga
     1021 gaatgaccga gtggagttgc ccagccgtac cctctgccag gccacccgag gcccctgtgc
     1081 cattgtggag cgggagcgag ggtggcctga ctttctgcgt tgcacaccgg accacttccc
     1141 tgaaggctgc ccaaacgagg tacaaaacat caagttcaac agctcaggcc aatgtgaagc
     1201 acccttggtg cgaacagaca accccaagag ctggtatgag gacgtggagg gctgtgggat
     1261 tcagtgtcag aacccgctgt tcaccgaggc cgagcaccag gacatgcaca gctacatcgc
     1321 agccttcggg gcggtcactg gtctctgcac gctcttcaca ctggccacct ttgtggctga
     1381 ctggcggaat tccaatcgct accctgcggt tattctcttc tatgtcaatg cgtgtttctt
     1441 cgtgggcagc attggctggc tggcccagtt catggatggt gcccgccgag agattgtttg
     1501 ccgagcagat ggcaccatga gatttgggga gcccacctcc agtgagaccc tgtcctgtgt
     1561 catcatcttt gtcattgtgt actatgcctt gatggctgga gtagtctggt tcgtggtcct
     1621 cacctatgcc tggcacacct ccttcaaagc cctgggcacc acctaccagc ctctctcggg
     1681 caagacatcc tatttccacc tgctcacgtg gtcactcccc tttgtcctca cggtggcaat
     1741 cctggctgtg gctcaggtag atggagactc cgtgagtggc atctgttttg taggctacaa
     1801 gaactatcgg taccgtgctg gctttgtcct ggccccaatt ggcctggtgc ttattgtggg
     1861 aggctacttc ctcatcagag gggtcatgac tctgttctcc atcaagagca accaccctgg
     1921 gctcttaagt gagaaggcag ccagcaagat caacgagacc atgctgcgcc tgggcatttt
     1981 tggcttcctg gcctttggct ttgtgctcat caccttcagc tgccacttct atgacttctt
     2041 caaccaggct gagtgggagc gtagcttccg ggactatgtg ctatgccaag ccaacgtgac
     2101 catcgggctg cctaccaaga agcccattcc tgactgtgag atcaagaatc ggcccagcct
     2161 cctggtggag aagatcaatc tatttgccat gtttggcact ggcattgcca tgagcacctg
     2221 ggtctggacc aaggccaccc tgctcatctg gaggcgcacc tggtgcaggt tgactgggca
     2281 cagtgatgat gagcccaaga gaatcaagaa gagcaagatg atcgccaagg ccttctctaa
     2341 gcggcgtgag ctgctgcaga acccgggcca ggagctctcc ttcagcatgc acactgtctc
     2401 ccatgatgga cctgttgcgg gtttggcttt tgacctcaat gaaccctcag ctgacgtctc
     2461 ctctgcctgg gctcagcatg tcaccaagat ggtggctcgg agaggagcca tattgcccca
     2521 ggatgtgtcc gttacccctg tggcaactcc agtgccacca gaagaacaag ccaacatgtg
     2581 gctggttgag gcagagatct ccccagagtt agagaagcgt ttgggccgga agaaaaagcg
     2641 gaggaagagg aagaaggagg tgtgcccctt ggggccagcc cctgagcttc accactctgc
     2701 ccctgttcct gccaccagtg cagttcctcg gctgcctcag ctgcctcggc agaagtgcct
     2761 ggtagctgca aacgcctggg gaacagggga gtcctgccga cagggagcct ggactctagt
     2821 ctccaacccc ttctgcccag agcctagtcc ccatcaagat ccatttctcc ctggtgcctc
     2881 agccccccgg gtctgggctc agggccgcct ccaggggctg ggatccattc attcccgcac
     2941 taacctaatg gaggctgaga tcttggatgc agactcggac ttcagaaaag agctagcgct
     3001 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     3061 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     3121 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     3181 gccctggccc accctcgtga ccaccctgac ctggggcgtg cagtgcttcg cccgctaccc
     3241 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     3301 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     3361 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     3421 catcctgggg cacaagctgg agtacaacgc catcagcgac aacgtctata tcaccgccga
     3481 caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcag
     3541 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     3601 gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg
     3661 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     3721 gctgtacaag tccggactca gatctcgagc tcaagcttcg aattctgcag tcgacggtac
     3781 cgcgggcccg ggatccaccg gatctagata actgatcata atcagccata ccacatttgt
     3841 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     3901 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     3961 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4021 caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     4081 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     4141 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     4201 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     4261 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     4321 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     4381 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     4441 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     4501 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4561 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4621 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     4681 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4741 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4801 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4861 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4921 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4981 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     5041 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     5101 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     5161 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     5221 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     5281 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     5341 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     5401 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5461 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5521 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     5581 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5641 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5701 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5761 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5821 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5881 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5941 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     6001 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     6061 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     6121 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     6181 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     6241 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     6301 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6361 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6421 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6481 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6541 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6601 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6661 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6721 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6781 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6841 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6901 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6961 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7021 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     7081 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
