LOCUS       sequence-96298-e        6619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a fusion protein of FRB-linker peptide 2- Cre(60-343).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6619)
  AUTHORS   Jullien N, Sampieri F, Enjalbert A, Herman JP
  TITLE     Regulation of Cre recombinase by ligand-induced complementation of 
            inactive fragments.
  JOURNAL   Nucleic Acids Res. 2003 Nov 1;31(21):e131.
  PUBMED    14576331
REFERENCE   2  (bases 1 to 6619)
  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: "Nucleic 
            Acids Res."; date: "2003-11-1"; volume: "31(21)"; pages: "e131"
COMMENT     Sequence Label: pcDNA_DiCre_60.F2
FEATURES             Location/Qualifiers
     source          1..6619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      958..967
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             970..990
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             997..1272
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRIS"
     primer_bind     complement(2213..2230)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2219..2443
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2489..2917
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2576..2595)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2786..2807
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2926..2946)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2931..3260
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3111..3246
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3173..3192
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3327..4121
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3381..3400)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3991..4010
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4295..4416
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4332..4351)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4386..4405
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4465..4481)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4465..4481)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4478..4500)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4489..4505
                     /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(4513..4519,4520..4537,4538..4543))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4558..4579
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4696..4713)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4867..5452)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4947..4966)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5623..6414,6415..6483))
                     /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     6246..6265
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6484..6588)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagg
      901 atcgatccac tagtccagtg tggtggaatt ctgcagatat ccagcacagt ggcggccgcc
      961 accatggccc ctaagaagaa gagaaaggtg tctagaatcc tctggcatga gatgtggcat
     1021 gaaggcctgg aagaggcatc tcgtttgtac tttggggaaa ggaacgtgaa aggcatgttt
     1081 gaggtgctgg agcccttgca tgctatgatg gaacggggcc cccagactct gaaggaaaca
     1141 tcctttaatc aggcctatgg tcgagattta atggaggccc aagagtggtg caggaagtac
     1201 atgaaatcag ggaatgtcaa ggacctcctc caagcctggg acctctatta tcatgtgttc
     1261 cgacgaatct cagctagccc cagcaacccc ggcgccagca acggatccaa caggaaatgg
     1321 ttccctgctg aacctgagga tgtgagggac tacctcctgt acctgcaagc cagaggcctg
     1381 gctgtgaaga ccatccaaca gcacctgggc cagctcaaca tgctgcacag gagatctggc
     1441 ctgcctcgcc cttctgactc caatgctgtg tccctggtga tgaggagaat cagaaaggag
     1501 aatgtggatg ctggggagag agccaagcag gccctggcct ttgaacgcac tgactttgac
     1561 caagtcagat ccctgatgga gaactctgac agatgccagg acatcaggaa cctggccttc
     1621 ctgggcattg cctacaacac cctgctgcgc attgccgaaa ttgccagaat cagagtgaag
     1681 gacatctccc gcaccgatgg tgggagaatg ctgatccaca ttggcaggac caagaccctg
     1741 gtgtccacag ctggtgtgga gaaggccctg tccctggggg ttaccaagct ggtggagaga
     1801 tggatctctg tgtctggtgt ggctgatgac cccaacaact acctgttctg ccgggtcaga
     1861 aagaatggtg tggctgcccc ttctgccacc tcccaactgt ccacccgggc cctggaaggg
     1921 atctttgagg ccacccaccg cctgatctat ggtgccaagg atgactctgg gcagagatac
     1981 ctggcctggt ctggccactc tgccagagtg ggtgctgcca gggacatggc cagggctggt
     2041 gtgtccatcc ctgaaatcat gcaggctggt ggctggacca atgtgaacat tgtgatgaac
     2101 tacatcagaa acctggactc tgagactggg gccatggtga ggctgctcga ggatggggac
     2161 tgagatctgg ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     2221 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2281 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2341 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2401 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2461 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2521 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2581 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2641 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2701 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2761 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2821 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2881 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     2941 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3001 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3061 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3121 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3181 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3241 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3301 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3361 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3421 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3481 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3541 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3601 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3661 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3721 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3781 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3841 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3901 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3961 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4021 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4081 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4141 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4201 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4261 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4321 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4381 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4441 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4501 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4561 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4621 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4681 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4741 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4801 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4861 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4921 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4981 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5041 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5101 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5161 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5221 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5281 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5341 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5401 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5461 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5521 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5581 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5641 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5701 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5761 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5821 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5881 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5941 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6001 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6061 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6121 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6181 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6241 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6301 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6361 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6421 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6481 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6541 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6601 aaaagtgcca cctgacgtc
//
