LOCUS       sequence-64918-e        6131 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 6131)
  AUTHORS   Ni TT, Lu J, Zhu M, Maddison LA, Boyd KL, Huskey L, Ju B, Hesselson 
            D, Zhong TP, Page-McCaw PS, Stainier DY, Chen W
  TITLE     Conditional control of gene function by an invertible gene trap in 
            zebrafish.
  JOURNAL   Proc Natl Acad Sci U S A. 2012 Sep 18;109(38):15389-94. Epub 2012 
            Aug 20.
  PUBMED    22908272
REFERENCE   2  (bases 1 to 6131)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2012-09-18"; volume: "109(38)"; pages: 
            "15389-94. Epub 2012 Aug 20"
COMMENT     Sequence Label: pFT1.0
FEATURES             Location/Qualifiers
     source          1..6131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(34..462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     356..375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     589..611
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     603..620
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     604..620
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     630..649
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        630..648
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(679..690,691..828)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     protein_bind    complement(846..879)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(926..959)
                     /label=lox5171
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTGTAC) (Shaw et al., 2021). lox5171 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    986..1019
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    1065..1098
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     polyA_signal    1126..1350
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    1433..1657
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    1700..1924
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    2051..2275
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    2318..2542
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(2583..3293)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2861..2880)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3130..3148
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    complement(3403..3450)
                     /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)."
     protein_bind    complement(3475..3508)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     protein_bind    3541..3574
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3621..3654
                     /label=lox5171
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTGTAC) (Shaw et al., 2021). lox5171 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     misc_feature    join(3661..3848,3849..3860)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3894..3910)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3894..3910)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3947..3967)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3947..3965)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3986..4002)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3986..4002)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3999..4021)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4010..4026
                     /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(4034..4040,4041..4058,4059..4064))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4079..4100
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4217..4234)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4388..4976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4468..4487)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5147..5938,5939..6007))
                     /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     5770..5789
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6008..6112)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
      241 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
      301 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
      361 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
      421 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
      481 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
      541 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
      601 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta
      661 ccggcatatg gttcttgaca gaggtgtaaa aagtactcaa aaattttact caagtgaaag
      721 tacaagtact tagggaaaat tttactcaat taaaagtaaa agtatctggc tagaatctta
      781 cttgagtaaa agtaaaaaag tactccatta aaattgtact tgagtattag atccgataag
      841 ctagcataac ttcgtatagc atacattata cgaagttatc caagcttcac catcgacccg
      901 aattgccaag catcaccatc gacccataac ttcgtatagt acacattata cgaagttatt
      961 cgctacctta ggaccgttat agttagaagt tcctattctc tagaaagtat aggaacttct
     1021 cattctggcc tccccctccc ccaaggccag gaagttccta ttccgaagtt cctattcttc
     1081 aaatagtata ggaacttcgg atcctggttc tttccgcctc agaagccata gagcccaccg
     1141 catccccagc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc
     1201 caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc tcattttatt
     1261 aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa
     1321 caacagatgg ctggcaacta gaaggcacag tcgaggctga tcagcggttt aaacttaaac
     1381 tccattaaaa ttgtacttga gtattagatc ctggttcttt ccgcctcaga agccatagag
     1441 cccaccgcat ccccagcatg cctgctattg tcttcccaat cctccccctt gctgtcctgc
     1501 cccaccccac cccccagaat agaatgacac ctactcagac aatgcgatgc aatttcctca
     1561 ttttattagg aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag
     1621 gggcaaacaa cagatggctg gcaactagaa ggcacagtcg aggctgatca gcggtttctg
     1681 gttctttccg cctcagaagc catagagccc accgcatccc cagcatgcct gctattgtct
     1741 tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta
     1801 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct
     1861 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc
     1921 acagtcgagg ctgatcagcg agctccaccg cggtggcggc cgctttactt gtacagctcg
     1981 tccatgccaa acttaaactc cattaaaatt gtacttgagt attagatcct ggttctttcc
     2041 gcctcagaag ccatagagcc caccgcatcc ccagcatgcc tgctattgtc ttcccaatcc
     2101 tcccccttgc tgtcctgccc caccccaccc cccagaatag aatgacacct actcagacaa
     2161 tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc
     2221 aaggaaggca cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagtcgag
     2281 gctgatcagc ggtttctggt tctttccgcc tcagaagcca tagagcccac cgcatcccca
     2341 gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc
     2401 agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg
     2461 acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
     2521 ggctggcaac tagaaggcac agtcgaggct gatcagcgag ctccaccgcg gtggcggccg
     2581 ctttacttgt acagctcgtc catgccgccg gtggagtggc ggccctcggc gcgttcgtac
     2641 tgttccacga tggtgtagtc ctcgttgtgg gaggtgatgt ccaacttgat gttgacgttg
     2701 taggcgccgg gcagctgcac gggcttcttg gccttgtagg tggtcttgac ctcagcgtcg
     2761 tagtggccgc cgtccttcag cttcagcctc tgcttgatct cgcccttcag ggcgccgtcc
     2821 tcggggtaca tccgctcgga ggaggcctcc cagcccatgg tcttcttctg cattacgggg
     2881 ccgtcggagg ggaagttggt gccgcgcagc ttcaccttgt agatgaactc gccgtcctgc
     2941 agggaggagt cctgggtcac ggtcaccacg ccgccgtcct cgaagttcat cacgcgctcc
     3001 cacttgaagc cctcggggaa ggacagcttc aagtagtcgg ggatgtcggc ggggtgcttc
     3061 acgtaggcct tggagccgta catgaactga ggggacagga tgtcccaggc gaagggcagg
     3121 gggccaccct tggtcacctt cagcttggcg gtctgggtgc cctcgtaggg gcggccctcg
     3181 ccctcgccct cgatctcgaa ctcgtggccg ttcacggagc cctccatgtg caccttgaag
     3241 cgcatgaact ccttgatgat ggccatgtta tcctcctcgc ccttgctcac catggtggtt
     3301 cttctcttct tccgaccgcg aagagtttgt cgatcgacct gaaaaaaaaa agggaagaga
     3361 gagacacgtc agaaacacac acacactccg gattagtctc gagaagttcc tatactttct
     3421 agagaatagg aacttcggaa taggaacttc acacacacac acacacacac acacgaagtt
     3481 cctatactat ttgaagaata ggaacttcgg aataggaact tcattaccct gttatcccta
     3541 ataacttcgt ataatgtatg ctatacgaag ttatccaagc atcaccatcg accctctagt
     3601 ccagatctca ccatcgaccc ataacttcgt ataatgtgta ctatacgaag ttatgggccc
     3661 aagtgatctc caaaaaataa gtactttttg actgtaaata aaattgtaag gagtaaaaag
     3721 tacttttttt tctaaaaaaa tgtaattaag taaaagtaaa agtattgatt tttaattgta
     3781 ctcaagtaaa gtaaaaatcc ccaaaaataa tacttaagta cagtaatcaa gtaaaattac
     3841 tcaagtactt tacacctctg gttcttgacc ccctaccttc agcaagccca gcagatccac
     3901 tagttctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt
     3961 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4021 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4081 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4141 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4201 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4261 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4321 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4381 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4441 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4501 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4561 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4621 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4681 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4741 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4801 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     4861 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4921 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4981 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5041 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5101 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5161 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5221 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5281 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5341 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5401 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5461 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5521 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5581 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5641 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5701 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5761 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5821 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5881 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5941 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6001 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6061 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6121 cccgaaaagt g
//
