LOCUS       sequence-54502-e        7198 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 7198)
  AUTHORS   Dorn KV, Hughes CE, Rohatgi R
  TITLE     A Smoothened-Evc2 Complex Transduces the Hedgehog Signal at Primary 
            Cilia.
  JOURNAL   Dev Cell. 2012 Sep 12. pii: S1534-5807(12)00323-1. doi: 
            10.1016/j.devcel.2012.07.004.
  PUBMED    22981989
REFERENCE   2  (bases 1 to 7198)
  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: "Dev Cell. 
            2012 Sep 12. pii: S1534-5807(12)00323-1. doi: 
            10.1016/j.devcel.2012.07.004."
COMMENT     Sequence Label: pCS2+ YFP-Smo
FEATURES             Location/Qualifiers
     source          1..7198
                     /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             join(259..261,262..264,265..975)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(304..325)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(565..584)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     912..933
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3247..3381
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3271..3290)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3325..3344
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3494..3514)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3494..3512)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3533..3549)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3533..3549)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3546..3568)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3557..3573
                     /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(3581..3587,3588..3605,3606..3611))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3626..3647
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3764..3781)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3935..4523)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4015..4034)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4694..5485,5486..5554))
                     /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     5317..5336
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5555..5659)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5685..6140)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5822..5843)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6034..6053
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6214..7198
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattca ccatggccgc tggccgcccc
      121 gtgcgtgggc ccgagctggc gccccggagg ctgctgcagt tgcttctgct ggtactgctg
      181 gggggcccgg gccggggggc ggccttgagc gggaacgtga ccgggcctgg gcctcacagc
      241 gccagcggga gctcgagcat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      301 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      361 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      421 cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc
      481 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      541 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
      601 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
      661 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
      721 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
      781 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
      841 ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag
      901 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
      961 gacgagctgt acaagagctc gaggagggac gtgccggtga ccagccctcc gccgccgctg
     1021 ctgagccact gcggccgggc cgcccactgc gagcctttgc gctacaacgt gtgcctgggc
     1081 tcggcgctgc cctacggagc caccaccacg ctgctggctg gggactcgga ctcgcaggag
     1141 gaagcgcacg gcaagctcgt gctctggtcc ggcctccgga atgccccccg ctgctgggca
     1201 gtgatccagc ccctgctgtg tgctgtctac atgcccaagt gtgagaatga ccgagtggag
     1261 ttgcccagcc gtaccctctg ccaggccacc cgaggcccct gtgccattgt ggagcgggag
     1321 cgagggtggc ctgactttct gcgttgcaca ccggaccact tccctgaagg ctgcccaaac
     1381 gaggtacaaa acatcaagtt caacagctca ggccaatgtg aagcaccctt ggtgcgaaca
     1441 gacaacccca agagctggta tgaggacgtg gagggctgtg ggattcagtg tcagaacccg
     1501 ctgttcaccg aggccgagca ccaggacatg cacagctaca tcgcagcctt cggggcggtc
     1561 actggtctct gcacgctctt cacactggcc acctttgtgg ctgactggcg gaactccaat
     1621 cgctaccctg cggttattct cttctatgtc aatgcgtgtt tcttcgtggg cagcattggc
     1681 tggctggccc agttcatgga tggtgcccgc cgagagattg tttgccgagc agatggcacc
     1741 atgagatttg gggagcccac ctccagtgag accctgtcct gtgtcatcat ctttgtcatt
     1801 gtgtactatg ccttgatggc tggagtagtc tggttcgtgg tcctcaccta tgcctggcac
     1861 acctccttca aagccctggg caccacctac cagcctctct cgggcaagac atcctatttc
     1921 cacctgctca cgtggtcact cccctttgtc ctcacggtgg caatcctggc tgtggctcag
     1981 gtagatggag actccgtgag tggcatctgt tttgtaggct acaagaacta tcggtaccgt
     2041 gctggctttg tcctggcccc aattggcctg gtgcttattg tgggaggcta cttcctcatc
     2101 agaggggtca tgactctgtt ctccatcaag agcaaccacc ctgggcttct gagtgagaag
     2161 gcagccagca agatcaacga gaccatgctg cgcctgggca tttttggctt cctggccttt
     2221 ggctttgtgc tcatcacctt cagctgccac ttctatgact tcttcaacca ggctgagtgg
     2281 gagcgtagct tccgggacta tgtgctatgc caagccaacg tgaccatcgg gctgcctacc
     2341 aagaagccca ttcctgactg tgagatcaag aatcggccca gcctcctggt ggagaagatc
     2401 aatctatttg ccatgtttgg cactggcatt gccatgagca cctgggtctg gaccaaggcc
     2461 accctgctca tctggaggcg cacctggtgc aggttgactg ggcacagtga tgatgagccc
     2521 aagagaatca agaagagcaa gatgatcgcc aaggccttct ctaagcggcg tgagctgctg
     2581 cagaacccgg gccaggagct ctccttcagc atgcacactg tctcccatga tggacctgtt
     2641 gcgggtttgg cttttgacct caatgaaccc tcagctgacg tctcctctgc ctgggctcag
     2701 catgtcacca agatggtggc tcggagagga gccatattgc cccaggatgt gtccgttacc
     2761 cctgtggcaa ctccagtgcc accagaagaa caagccaaca tgtggctggt tgaggcagag
     2821 atctccccag agttagagaa gcgtttgggc cggaagaaaa agcggaggaa gaggaagaag
     2881 gaggtgtgcc ccttgaggcc agcccctgag cttcaccact ctgcccctgt tcctgccacc
     2941 agtgcagttc ctcggctgcc tcagctgcct cggcagaagt gcctggtagc tgcaaacgcc
     3001 tggggaacag gggagtcctg ccgacaggga gcctggactc tagtctccaa ccccttctgc
     3061 ccagagccta gtccccatca agatccattt ctccctggtg cctcagcccc ccgggtctgg
     3121 gctcagggcc gcctccaggg gctgggatcc attcattccc gcactaacct aatggaggct
     3181 gagatcttgg atgcagactc ggacttcggc gcgccagtct agaactatag tgagtcgtat
     3241 tacgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     3301 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     3361 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     3421 ggggaggtgt gggaggtttt ttaattcgcg gccgcggcgc caatgcattg ggcccggtac
     3481 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     3541 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3601 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     3661 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3721 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3781 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3841 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3901 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3961 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4021 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4081 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4141 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4201 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4261 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4321 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     4381 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4441 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4501 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4561 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4621 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4681 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4741 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4801 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4861 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4921 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4981 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5041 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5101 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5161 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5221 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5281 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5341 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5401 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5461 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5521 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5581 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5641 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
     5701 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     5761 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     5821 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     5881 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     5941 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     6001 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     6061 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     6121 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
     6181 atcggtgcgg gcctcttcgc tattacgcca gtcgaccata gccaattcaa tatggcgtat
     6241 atggactcat gccaattcaa tatggtggat ctggacctgt gccaattcaa tatggcgtat
     6301 atggactcgt gccaattcaa tatggtggat ctggacccca gccaattcaa tatggcggac
     6361 ttggcaccat gccaattcaa tatggcggac ttggcactgt gccaactggg gaggggtcta
     6421 cttggcacgg tgccaagttt gaggaggggt cttggccctg tgccaagtcc gccatattga
     6481 attggcatgg tgccaataat ggcggccata ttggctatat gccaggatca atatataggc
     6541 aatatccaat atggccctat gccaatatgg ctattggcca ggttcaatac tatgtattgg
     6601 ccctatgcca tatagtattc catatatggg ttttcctatt gacgtagata gcccctccca
     6661 atgggcggtc ccatatacca tatatggggc ttcctaatac cgcccatagc cactccccca
     6721 ttgacgtcaa tggtctctat atatggtctt tcctattgac gtcatatggg cggtcctatt
     6781 gacgtatatg gcgcctcccc cattgacgtc aattacggta aatggcccgc ctggctcaat
     6841 gcccattgac gtcaatagga ccacccacca ttgacgtcaa tgggatggct cattgcccat
     6901 tcatatccgt tctcacgccc cctattgacg tcaatgacgg taaatggccc acttggcagt
     6961 acatcaatat ctattaatag taacttggca agtacattac tattggaagg acgccagggt
     7021 acattggcag tactcccatt gacgtcaatg gcggtaaatg gcccgcgatg gctgccaagt
     7081 acatccccat tgacgtcaat ggggaggggc aatgacgcaa atgggcgttc cattgacgta
     7141 aatgggcggt aggcgtgcct aatgggaggt ctatataagc aatgctcgtt tagggaac
//
