LOCUS       sequence-11449-e        7960 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 7960)
  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 7960)
  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. 
            2010 Mar 16. 18(3):410-24."
COMMENT     Sequence Label: pFloxin-IRES-Ofd1-Myc-S75F
FEATURES             Location/Qualifiers
     source          1..7960
                     /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)."
     misc_feature    876..1448
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    899..1450
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1043..1060)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             4739..4768
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    5112..5246
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5149..5168)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5203..5222
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    5264..5311
                     /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(5335..5351)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5335..5351)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5348..5370)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5359..5375
                     /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(5383..5389,5390..5407,5408..5413))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5428..5449
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5566..5583)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5737..6325)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5817..5836)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6496..7287,7288..7356))
                     /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     7119..7138
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7357..7461)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7529..7547
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7585..7607)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7707..7726
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7920..7942
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7934..7951
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7935..7951
                     /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 cacccccagt atctgcaacc tcaagctagc ttgggtgcgt tggttgtgga taagtagcta
      781 gactccagca accagtaacc tctgcccttt ctcctccatg acaaccaggt accggtagtg
      841 actaatagct gatagaattc cgcccccccc ccccccccct ctccctcccc cccccctaac
      901 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
      961 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     1021 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     1081 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     1141 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     1201 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     1261 agagtcaaat ggctctccta agcgtattca acaaggggct gaaggatgcc cagaaggtac
     1321 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     1381 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     1441 gatgataata ggatcgcatg cggcgcgcca ctagcgctac cggactcaga tctcgagctc
     1501 aagcttcgaa ttatctgcag aattcgccct tgacgagtta ggggctgaat gtttagctct
     1561 cttgctcccc taggtcccca gagacaagct ggattccgag gtcaagggct ccctctcctc
     1621 ctgcgccccg gcccgggccg aggagtccga ccgtggaggg gcgagctcgg gaggaaagca
     1681 gatgaggatg gctcagtcca acatgcccca caagtctgat gtgttgagtc aagatgaact
     1741 gcgcaaaaag ctataccaga cgtttaagga tcggggtgta ctggacacac ttcagaccca
     1801 actccgaaac cagctaattc atgaattgat gcatcctgta ttgagtggag aagtgaagcc
     1861 gccatccatt tcagtggaag ggagtgccct cctaataggg gcttttaact cgctggtggc
     1921 agaccactta caaagatgtg gctatgaata ttcactctct gttttctttc cagaaagtgg
     1981 tttggcaaaa gaaaagatat ttactatgca ggatctacta caactcattc ggattaaccc
     2041 ctcatccagt ctctataaat cattgatttc aggatttgat aaagaaaata aaaaaggttt
     2101 tcttatgagt ttcttaaagg agttggcaga atattatcaa gctaaagaga gctgtgatgc
     2161 ggaaactcag acaagcacca catttcccag ccaagtgtcc ctagccgaga agtttcagct
     2221 tattgacgct caatttgcag atggttttcc tcaccgttcc aagctagaat ctttagagac
     2281 aaagttaaac gagtataaga aagaagtaca acaccagctt caggtagaaa tgtgtcataa
     2341 gctgaagtat tttagagaag ctgaaataac aaaagttaag atggaagaga gaagaaagta
     2401 tgaaaaggaa ttggctgagt tccagaatga atttgaaaga acttgtcaag caaaaaatga
     2461 agccctcatt tctcaggaga agaattctct ggaaagaatt aaaaaacacc gagagatgga
     2521 atcaaaagaa atttatgctc aaagacagct tctactaaat gatatagctt tgcttagagg
     2581 tagagaagca gagctgaagg agcgaatcga aacttttgaa ttgacccaga agctccaaga
     2641 agaaaaaatt aaaagtgagg ctgaggcact tgagagacgg gaacagaatc tgaagaatat
     2701 tgaagacacc tatgatcaga aactcaaaac tgaactgcta aagtaccaac tggagctaaa
     2761 ggatgactac atcactagga caaataaact cctggaagag gaaaggaaga ataaagaaaa
     2821 aactattcat ttgcaagagg agctcacagt tattaattca aaaaaggaag aactcagtaa
     2881 atctgttaag catatgaaag aagttgagct ggaactggag tctgtgaaag ctcagttttt
     2941 agcaatatca aaacaaaacc acttgctgaa tgaaaaggtg agagagatga gtgattactc
     3001 acaactgaaa gaagagaaag tggagcttca ggcacaaaat aaactactta aactgcaact
     3061 ggaagagact aggaatgaaa acttacgact cctagaccgc ataactcagc cacctcctga
     3121 gcttgtgatt tttcagaagg aattacagaa aacagaaaag gctatggaac tggagcacaa
     3181 ggactttgaa actcataggc aggctcttga gaaacagctg cagagtgaaa ttgaaaattc
     3241 tgcacagcta aggacccaga tagcagaata tgatgcttct gtcaaaaggt taactgttca
     3301 ggtggctgag ttaaagtcac aactaaaaca aactcagata gccctagaga atgaagtgta
     3361 ccgaaatcca aaacactctc tgattcattc cctgagtggc ctcctattga gcggcaagat
     3421 ggcaccccac agtgaagata aaagtgggga tttcttgaat gtaccccttg aacagaacaa
     3481 ggttatagcg ggtgcagtta tgtcaagggt cccaccttat gtaaatacag caacagaagc
     3541 tagttcccca gagtctgact ttgagtttat agccagtagt actaaagcca aagtaagaga
     3601 gctagagcaa gaggctgaac gcttggaaaa ggcattcaga acttactatc aaagagctac
     3661 tcagaatcct tctacgagcc ctcagccggc aaagagtcct ccatctgtaa actcggtggc
     3721 agctctcaga agcatagctt ccagttccat ggacagacct gtttctgcag aggacagagt
     3781 tgtctcggaa cagcctcttg gggacatgct gaaggaagaa atgagtgata tgtccaaagc
     3841 attcatgggc agtgtggttt cacggccacg caggacctct tcctccacac gcctctcctc
     3901 tacaccccat ccaaaatcga gaagaagtct tgataatgaa atgtatctgg aaggtctcgg
     3961 gagattacac atgacttcct ccagccctct tcttgacaga gtttctgcat cacctgctgc
     4021 ttcccctagt ccttgtcctg agagaacagc ccaggcgtca cctgttccat ctaggcacag
     4081 cttctctggt cttccagagc aaaatgcctg tttatatcag agacaaactg aaacgcaaga
     4141 taaaagtgag ctttcaaatg tggacaagca atctttgaaa gatgagaaat ttgaaccacc
     4201 tttcagatgg aacaagacag aacagtttga agcagaagga ctccaccctg ctggtgacat
     4261 gcctggaata gattttgctg tggctactca gtcttcaaga ctcatttcat acgactaccc
     4321 aagtgctgtt cagagccaaa ctggagaaca agacgaacag gagctatggg agcttcacat
     4381 gaaagaacgc aggcagagag aagaacaaag gcataatgaa cggcaagaag ctttagagag
     4441 ggaaagaaga gagctgggaa aactagagca ggagcggagg atgattgaag agtcactgaa
     4501 gatggagatg gaagaagaat tagaaaagag tgttcaagac cagaaagaca aatcagctca
     4561 ttgtgaaaat actttagaaa aatacatgaa aattatccag cagagacagg aagaatcaaa
     4621 tgcagataag agctccaaaa aatcgggcaa agaatgctca ctagtggaca tgatgatgcc
     4681 tagtgacaag gatgaaagtt ctccaggatt ttctcatgaa gaaccagatg acatgtggga
     4741 gcaaaagctc atttctgaag aggacttgta aaagggcgaa ttccagcaca ctggcggccg
     4801 ttactagtgg atccgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt
     4861 ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg
     4921 caatgtgaga tcgctagcgg atccaccacc tacagcccag tggggtcgct ctagagcgac
     4981 ctcgaggggg ggctagataa gtaatgatca taatcagcca tatcacatct gtagaggttt
     5041 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     5101 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5161 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5221 tcaatgtatc ttatcatgtc tagatccggg ggtaccgcgt cgagaagttc ctattccgaa
     5281 gttcctattc tctagaaagt ataggaactt cgtcgacgcg aattcgtaat catggtcata
     5341 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5401 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5461 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5521 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5581 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5641 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5701 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5761 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5821 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5881 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5941 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6001 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6061 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6121 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6181 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6241 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6301 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6361 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6421 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6481 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6541 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6601 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6661 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6721 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6781 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6841 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6901 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6961 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7021 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7081 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7141 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7201 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7261 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7321 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7381 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7441 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     7501 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     7561 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     7621 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     7681 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     7741 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     7801 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     7861 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     7921 ccagtcacga cgttgtaaaa cgacggccag tgccaagctt
//
