LOCUS       sequence-54503-e        7329 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 7329)
  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 7329)
  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-Weyer's
FEATURES             Location/Qualifiers
     source          1..7329
                     /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    3378..3512
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3402..3421)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3456..3475
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3625..3645)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3625..3643)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3664..3680)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3664..3680)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3677..3699)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3688..3704
                     /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(3712..3718,3719..3736,3737..3742))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3757..3778
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3895..3912)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4066..4654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4146..4165)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4825..5616,5617..5685))
                     /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     5448..5467
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5686..5790)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5816..6271)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5953..5974)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6165..6184
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6345..7329
                     /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 gcgccattaa acacgggaga aaagatcttt
     3241 gtattcagaa gcccaaggga gcccgagatc tctttgcgtg ttcctcccag gaggaaaaag
     3301 aatttcctga atgccaagaa ggccaacagg gccttgggct tggactgatc tagaactata
     3361 gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca
     3421 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     3481 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     3541 ttcaggttca gggggaggtg tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt
     3601 gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     3661 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3721 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     3781 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3841 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3901 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3961 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4021 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4081 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4141 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4201 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4261 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4321 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4381 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4441 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4501 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4561 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4621 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4681 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4741 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4801 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4861 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4921 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4981 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5041 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5101 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5161 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5221 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5281 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5341 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5401 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5461 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5521 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5581 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5641 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5701 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5761 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     5821 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     5881 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     5941 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     6001 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     6061 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     6121 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     6181 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     6241 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     6301 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agtcgaccat agccaattca
     6361 atatggcgta tatggactca tgccaattca atatggtgga tctggacctg tgccaattca
     6421 atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc agccaattca
     6481 atatggcgga cttggcacca tgccaattca atatggcgga cttggcactg tgccaactgg
     6541 ggaggggtct acttggcacg gtgccaagtt tgaggagggg tcttggccct gtgccaagtc
     6601 cgccatattg aattggcatg gtgccaataa tggcggccat attggctata tgccaggatc
     6661 aatatatagg caatatccaa tatggcccta tgccaatatg gctattggcc aggttcaata
     6721 ctatgtattg gccctatgcc atatagtatt ccatatatgg gttttcctat tgacgtagat
     6781 agcccctccc aatgggcggt cccatatacc atatatgggg cttcctaata ccgcccatag
     6841 ccactccccc attgacgtca atggtctcta tatatggtct ttcctattga cgtcatatgg
     6901 gcggtcctat tgacgtatat ggcgcctccc ccattgacgt caattacggt aaatggcccg
     6961 cctggctcaa tgcccattga cgtcaatagg accacccacc attgacgtca atgggatggc
     7021 tcattgccca ttcatatccg ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc
     7081 cacttggcag tacatcaata tctattaata gtaacttggc aagtacatta ctattggaag
     7141 gacgccaggg tacattggca gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat
     7201 ggctgccaag tacatcccca ttgacgtcaa tggggagggg caatgacgca aatgggcgtt
     7261 ccattgacgt aaatgggcgg taggcgtgcc taatgggagg tctatataag caatgctcgt
     7321 ttagggaac
//
