LOCUS       sequence-11452-e        6802 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6802)
  AUTHORS   Singla V, Romaguera-Ros M, Garcia-Verdugo JM, Reiter JF
  TITLE     Ofd1, a human disease gene, regulates the length and distal 
            structure of centrioles.
  JOURNAL   Dev Cell. 2010 Mar 16. 18(3):410-24.
  PUBMED    20230748
REFERENCE   2  (bases 1 to 6802)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Dev Cell. 
            2010 Mar 16. 18(3):410-24."
COMMENT     Sequence Label: pFloxin-Ofd1-Myc-S437R
FEATURES             Location/Qualifiers
     source          1..6802
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      517..526
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(529..562)
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             3573..3602
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    3954..4088
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3991..4010)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4045..4064
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    4106..4153
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(4177..4193)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4177..4193)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4190..4212)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4201..4217
                     /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(4225..4231,4232..4249,4250..4255))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4270..4291
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4408..4425)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4579..5167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4659..4678)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5338..6129,6130..6198))
                     /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     5961..5980
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6199..6303)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6371..6389
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6427..6449)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6549..6568
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6762..6784
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6776..6793
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6777..6793
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aagcttcctt ctgcaggagc gtacagaacc cagggccctg gcacccgtgc agaccctggc
       61 ccaccccacc tgggcgctca gtgcccaaga gatgtccaca cctaggatgt cccgcggtgg
      121 gtggggggcc cgagagacgg gcaggccggg ggcaggcctg gccatgcggg gccgaaccgg
      181 gcactgccca gcgtggggcg cgggggccac ggcgcgcgcc cccagccccc gggcccagca
      241 ccccaaggcg gccaacgcca aaactctccc tcctcctctt cctcaatctc gctctcgctc
      301 tttttttttt tcgcaaaagg aggggagagg gggtaaaaaa atgctgcact gtcggcgaag
      361 ccggtgagtg agcggcgcgg ggccaatcgc gtgcgccgtt ccgaaagttg ccttttatgg
      421 ctcgagcggc cgcggcggcg ccctataaaa cccagcggcg cgacgcgcca ccaccgccga
      481 gaccgcgtcc gcccgcgagc acagagcctc gcctttgcca ccatggctat aacttcgtat
      541 agcatacatt atacgaacgg tagtgatagg ccttttagct acatctgcca atccatctca
      601 ttttcacaca cacacacacc actttccttc tggtcagtgg gcacatgtcc agcctcaagt
      661 ttatatcacc acccccaatg cccaacactt gtatggcctt gggcgggtca tccccccccc
      721 cccaccccca gtatctgcaa cctcaagcta gcttgggtgc gttggttgtg gataagtagc
      781 tagactccag caaccagtaa cctctgccct ttctcctcca tgacaaccag atatttacta
      841 tgcaggatct actacaactc attcggatta acccctcatc cagtctctat aaatcattga
      901 tttcaggatt tgataaagaa aataaaaaag gttttcttat gagtttctta aaggagttgg
      961 cagaatatta tcaagctaaa gagagctgtg atgcggaaac tcagacaagc accacatttc
     1021 ccagccaagt gtccctagcc gagaagtttc agcttattga cgctcaattt gcagatggtt
     1081 ttcctcaccg ttccaagcta gaatctttag agacaaagtt aaacgagtat aagaaagaag
     1141 tacaacacca gcttcaggta gaaatgtgtc ataagctgaa gtattttaga gaagctgaaa
     1201 taacaaaagt taagatggaa gagagaagaa agtatgaaaa ggaattggct gagttccaga
     1261 atgaatttga aagaacttgt caagcaaaaa atgaagccct catttctcag gagaagaatt
     1321 ctctggaaag aattaaaaaa caccgagaga tggaatcaaa agaaatttat gctcaaagac
     1381 agcttctact aaatgatata gctttgctta gaggtagaga agcagagctg aaggagcgaa
     1441 tcgaaacttt tgaattgacc cagaagctcc aagaagaaaa aattaaaagt gaggctgagg
     1501 cacttgagag acgggaacag aatctgaaga atattgaaga cacctatgat cagaaactca
     1561 aaactgaact gctaaagtac caactggagc taaaggatga ctacatcact aggacaaata
     1621 aactcctgga agaggaaagg aagaataaag aaaaaactat tcatttgcaa gaggagctca
     1681 cagttattaa ttcaaaaaag gaagaactca gtaaatctgt taagcatatg aaagaagttg
     1741 agctggaact ggagtctgtg aaagctcagt ttttagcaat atcaaaacaa aaccacttgc
     1801 tgaatgaaaa ggtgagagag atgcgtgatt actcacaact gaaagaagag aaagtggagc
     1861 ttcaggcaca aaataaacta cttaaactgc aactggaaga gactaggaat gaaaacttac
     1921 gactcctaga ccgcataact cagccacctc ctgagcttgt gatttttcag aaggaattac
     1981 agaaaacaga aaaggctatg gaactggagc acaaggactt tgaaactcat aggcaggctc
     2041 ttgagaaaca gctgcagagt gaaattgaaa attctgcaca gctaaggacc cagatagcag
     2101 aatatgatgc ttctgtcaaa aggttaactg ttcaggtggc tgagttaaag tcacaactaa
     2161 aacaaactca gatagcccta gagaatgaag tgtaccgaaa tccaaaacat tctctgattc
     2221 attccctgag tggcctccta ttgagcggca agatggcacc ccacagtgaa gataaaagtg
     2281 gggatttctt gaatgtaccc cttgaacaga acaaggttat agcgggtgca gttatgtcaa
     2341 gggtcccacc ttatgtaaat acagcaacag aagctagttc cccagagtct gactttgagt
     2401 ttatagccag tagtactaaa gccaaagtaa gagagctaga gcaagaggct gaacgcttgg
     2461 aaaaggcatt cagaacttac tatcaaagag ctactcagaa tccttctacg agccctcagc
     2521 cggcaaagag tcctccatct gtaaactcgg tggcagctct cagaagcata gcttccagtt
     2581 ccatggacag acctgtttct gcagaggaca gagttgtctc ggaacagcct cttggggaca
     2641 tgctgaagga agaaatgagt gatatgtcca aagcattcat gggcagtgtg gtttcacggc
     2701 cacgcaggac ctcttcctcc acacgcctct cctctacacc ccatccaaaa tcgagaagaa
     2761 gtcttgataa tgaaatgtat ctggaaggtc tcgggagatt acacatgact tcctccagcc
     2821 ctcttcttga cagagtttct gcatcacctg ctgcttcccc tagtccttgt cctgagagaa
     2881 cagcccaggc gtcacctgtt ccatctaggc acagcttctc tggtcttcca gagcaaaatg
     2941 cctgtttata tcagagacaa actgaaacgc aagataaaag tgagctttca aatgtggaca
     3001 agcaatcttt gaaagatgag aaatttgaac cacctttcag atggaacaag acagaacagt
     3061 ttgaagcaga aggactccac cctgctggtg acatgcctgg aatagatttt gctgtggcta
     3121 ctcagtcttc aagactcatt tcatacgact acccaagtgc tgttcagagc caaactggag
     3181 aacaagacga acaggagcta tgggagcttc acatgaaaga acgcaggcag agagaagaac
     3241 aaaggcataa tgaacggcaa gaagctttag agagggaaag aagagagctg ggaaaactag
     3301 agcaggagcg gaggatgatt gaagagtcac tgaagatgga gatggaagaa gaattagaaa
     3361 agagtgttca agaccagaaa gacaaatcag ctcattgtga aaatacttta gaaaaataca
     3421 tgaaaattat ccagcagaga caggaagaat caaatgcaga taagagctcc aaaaaatcgg
     3481 gcaaagaatg ctcactagtg gacatgatga tgcctagtga caaggatgaa agttctccag
     3541 gattttctca tgaagaacca gatgacatgt gggagcaaaa gctcatttct gaagaggact
     3601 tgtaaaaggg cgaattccag cacactggcg gccgttacta gtggatccat ccgcccctct
     3661 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     3721 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc catcgctagc
     3781 ggatccacca cctacagccc agtggggtcg ctctagagcg acctcgaggg ggggctagat
     3841 aagtaatgat cataatcagc catatcacat ctgtagaggt tttacttgct ttaaaaaacc
     3901 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     3961 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4021 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4081 tctagatccg ggggtaccgc gtcgagaagt tcctattccg aagttcctat tctctagaaa
     4141 gtataggaac ttcgtcgacg cgaattcgta atcatggtca tagctgtttc ctgtgtgaaa
     4201 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4261 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4321 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4381 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4441 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4501 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4561 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4621 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4681 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4741 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4801 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4861 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4921 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4981 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     5041 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5101 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5161 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5221 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5281 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5341 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5401 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5461 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5521 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5581 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5641 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5701 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5761 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5821 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5881 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5941 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6001 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6061 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6121 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6181 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6241 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6301 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     6361 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     6421 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     6481 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     6541 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     6601 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     6661 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     6721 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     6781 aacgacggcc agtgccaagc tt
//
