LOCUS       sequence-11444-e        8051 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 8051)
  AUTHORS   Singla V, Hunkapiller J, Santos N, Seol AD, Norman AR, Wakenight P, 
            Skarnes WC, Reiter JF
  TITLE     Floxin, a resource for genetically engineering mouse ESCs.
  JOURNAL   Nat Methods. 2010 Jan . 7(1):50-2.
  PUBMED    19966808
REFERENCE   2  (bases 1 to 8051)
  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: "Nat 
            Methods. 2010 Jan . 7(1):50-2."
COMMENT     Sequence Label: pFloxin-IRES-Ofd1-Myc
FEATURES             Location/Qualifiers
     source          1..8051
                     /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             4822..4851
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    5203..5337
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5240..5259)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5294..5313
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    5355..5402
                     /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(5426..5442)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5426..5442)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5439..5461)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5450..5466
                     /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(5474..5480,5481..5498,5499..5504))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5519..5540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5657..5674)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5828..6416)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5908..5927)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6587..7378,7379..7447))
                     /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     7210..7229
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7448..7552)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7620..7638
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7676..7698)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7798..7817
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8011..8033
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8025..8042
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8026..8042
                     /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 ttccgtcttt attccgtcca gagtccctcc
     1561 tggctcgcgg acccgcccac ccttcccata ccgtgggagg aagcgcggct gcttgacgag
     1621 ttaggggctg aatgtttagc tctcttgctc ccctaggtcc ccagagacaa gctggattcc
     1681 gaggtcaagg gctccctctc ctcctgcgcc ccggcccggg ccgaggagtc cgaccgtgga
     1741 ggggcgagct cgggaggaaa gcagatgagg atggctcagt ccaacatgcc ccacaagtct
     1801 gatgtgttga gtcaagatga actgcgcaaa aagctatacc agacgtttaa ggatcggggt
     1861 gtactggaca cacttcagac ccaactccga aaccagctaa ttcatgaatt gatgcatcct
     1921 gtattgagtg gagaagtgaa gccgccatcc atttcagtgg aagggagtgc cctcctaata
     1981 ggggcttcta actcgctggt ggcagaccac ttacaaagat gtggctatga atattcactc
     2041 tctgttttct ttccagaaag tggtttggca aaagaaaaga tatttactat gcaggatcta
     2101 ctacaactca ttcggattaa cccctcatcc agtctctata aatcattgat ttcaggattt
     2161 gataaagaaa ataaaaaagg ttttcttatg agtttcttaa aggagttggc agaatattat
     2221 caagctaaag agagctgtga tgcggaaact cagacaagca ccacatttcc cagccaagtg
     2281 tccctagccg agaagtttca gcttattgac gctcaatttg cagatggttt tcctcaccgt
     2341 tccaagctag aatctttaga gacaaagtta aacgagtata agaaagaagt acaacaccag
     2401 cttcaggtag aaatgtgtca taagctgaag tattttagag aagctgaaat aacaaaagtt
     2461 aagatggaag agagaagaaa gtatgaaaag gaattggctg agttccagaa tgaatttgaa
     2521 agaacttgtc aagcaaaaaa tgaagccctc atttctcagg agaagaattc tctggaaaga
     2581 attaaaaaac accgagagat ggaatcaaaa gaaatttatg ctcaaagaca gcttctacta
     2641 aatgatatag ctttgcttag aggtagagaa gcagagctga aggagcgaat cgaaactttt
     2701 gaattgaccc agaagctcca agaagaaaaa attaaaagtg aggctgaggc acttgagaga
     2761 cgggaacaga atctgaagaa tattgaagac acctatgatc agaaactcaa aactgaactg
     2821 ctaaagtacc aactggagct aaaggatgac tacatcacta ggacaaataa actcctggaa
     2881 gaggaaagga agaataaaga aaaaactatt catttgcaag aggagctcac agttattaat
     2941 tcaaaaaagg aagaactcag taaatctgtt aagcatatga aagaagttga gctggaactg
     3001 gagtctgtga aagctcagtt tttagcaata tcaaaacaaa accacttgct gaatgaaaag
     3061 gtgagagaga tgagtgatta ctcacaactg aaagaagaga aagtggagct tcaggcacaa
     3121 aataaactac ttaaactgca actggaagag actaggaatg aaaacttacg actcctagac
     3181 cgcataactc agccacctcc tgagcttgtg atttttcaga aggaattaca gaaaacagaa
     3241 aaggctatgg aactggagca caaggacttt gaaactcata ggcaggctct tgagaaacag
     3301 ctgcagagtg aaattgaaaa ttctgcacag ctaaggaccc agatagcaga atatgatgct
     3361 tctgtcaaaa ggttaactgt tcaggtggct gagttaaagt cacaactaaa acaaactcag
     3421 atagccctag agaatgaagt gtaccgaaat ccaaaacact ctctgattca ttccctgagt
     3481 ggcctcctat tgagcggcaa gatggcaccc cacagtgaag ataaaagtgg ggatttcttg
     3541 aatgtacccc ttgaacagaa caaggttata gcaggtgcag ttatgtcaag ggtcccacct
     3601 tatgtaaata cagcaacaga agctagttcc ccagagtctg actttgagtt tatagccagt
     3661 agtactaaag ccaaagtaag agagctagag caagaggctg aacgcttgga aaaggcattc
     3721 agaacttact atcaaagagc tactcagaat ccttctacga gccctcagcc ggcaaagagt
     3781 cctccatctg taaactcggt ggcagctctc agaagcatag cttccagttc catggacaga
     3841 cctgtttctg cagaggacag agttgtctcg gaacagcctc ttggggacat gctgaaggaa
     3901 gaaatgagtg atatgtccaa agcattcatg ggcagtgtgg tttcacggcc acgcaggacc
     3961 tcttcctcca cacgcctctc ctctacaccc catccaaaat cgagaagaag tcttgataat
     4021 gaaatgtatc tggaaggtct cgggagatta cacatgactt cctccagccc tcttcttgac
     4081 agagtttctg catcacctgc tgcttcccct agtccttgtc ctgagagaac agcccaggcg
     4141 tcacctgttc catctaggca cagcttctct ggtcttccag agcaaaatgc ctgtttatat
     4201 cagagacaaa ctgaaacgca agataaaagt gagctttcaa atgtggacaa gcaatctttg
     4261 aaagatgaga aatttgaacc acctttcaga tggaacaaga cagaacagtt tgaagcagaa
     4321 ggactccacc ctgctggtga catgcctgga atagattttg ctgtggctac tcagtcttca
     4381 agactcattt catacgacta cccaagtgct gttcagagcc aaactggaga acaagacgaa
     4441 caggagctat gggagcttca catgaaagaa cgcaggcaga gagaagaaca aaggcataat
     4501 gaacggcaag aagctttaga gagggaaaga agagagctgg gaaaactaga gcaggagcgg
     4561 aggatgattg aagagtcact gaagatggag atggaagaag aattagaaaa gagtgttcaa
     4621 gaccagaaag acaaatcagc tcattgtgaa aatactttag aaaaatacat gaaaattatc
     4681 cagcagagac aggaagaatc aaatgcagat aagagctcca aaaaatcggg caaagaatgc
     4741 tcactagtgg acatgatgat gcctagtgac aaggatgaaa gttctccagg attttctcat
     4801 gaagaaccag atgacatgtg ggagcaaaag ctcatttctg aagaggactt gtaaaagggc
     4861 gaattccagc acactggcgg ccgttactag tggatccatc cgcccctctc cctccccccc
     4921 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     4981 attttccacc atattgccgt cttttggcaa tgtgagggcc atcgctagcg gatccaccac
     5041 ctacagccca gtggggtcgc tctagagcga cctcgagggg gggctagata agtaatgatc
     5101 ataatcagcc atatcacatc tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     5161 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     5221 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5281 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctagatccgg
     5341 gggtaccgcg tcgagaagtt cctattccga agttcctatt ctctagaaag tataggaact
     5401 tcgtcgacgc gaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5461 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5521 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5581 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5641 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5701 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5761 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5821 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5881 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5941 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6001 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6061 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6121 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6181 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6241 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     6301 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6361 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6421 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6481 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6541 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6601 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6661 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6721 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6781 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6841 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6901 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6961 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7021 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7081 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7141 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7201 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7261 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7321 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7381 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7441 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7501 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7561 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     7621 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     7681 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     7741 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     7801 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     7861 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     7921 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     7981 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     8041 gtgccaagct t
//
