LOCUS       sequence-103217-        6474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of FLPo recombinase from the broadly active CAG
            promoter/enhancer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6474)
  AUTHORS   Ohtsuka M, Miura H, Mochida K, Hirose M, Hasegawa A, Ogura A, 
            Mizutani R, Kimura M, Isotani A, Ikawa M, Sato M, Gurumurthy CB
  TITLE     One-step generation of multiple transgenic mouse lines using an 
            improved Pronuclear Injection-based Targeted Transgenesis (i-PITT).
  JOURNAL   BMC Genomics. 2015 Apr 9;16:274. doi: 10.1186/s12864-015-1432-5.
  PUBMED    25887549
REFERENCE   2  (bases 1 to 6474)
  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: "BMC 
            Genomics."; date: "2015-04-9"; volume: "16:274. doi"; pages: " 
            10.1186/s12864-015-1432-5"
COMMENT     Sequence Label: pBCV
FEATURES             Location/Qualifiers
     source          1..6474
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        320..623
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        626..901
                     /label=chicken beta-actin promoter
     intron          902..1910
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1834..1856
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1918..1937
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1978..1987
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1984..1989,1990..2010,2011..3282)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     polyA_signal    3420..3541
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3457..3476)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3511..3530
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3603..3620)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3603..3619)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3612..3634)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3832..4287
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3919..3938)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4129..4150
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4304..4323)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4423..4445
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4483..4501)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4569..4673
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4674..4742,4743..5534)
                     /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     complement(4892..4911)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5705..6293
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6194..6213
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6447..6464
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 catgcctgca ggtcgactct agttattaat agtaatcaat tacggggtca ttagttcata
      301 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      361 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      421 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      481 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      541 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      601 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      661 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      721 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      781 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      841 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      901 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      961 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
     1021 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
     1081 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
     1141 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
     1201 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1261 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1321 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1381 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1441 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1501 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1561 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1621 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1681 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1741 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1801 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1861 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1921 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attgggatcc accggtcgcc
     1981 accatggctc ctaagaagaa gaggaaggtg atgagccagt tcgacatcct gtgcaagacc
     2041 ccccccaagg tgctggtgcg gcagttcgtg gagagattcg agaggcccag cggcgagaag
     2101 atcgccagct gtgccgccga gctgacctac ctgtgctgga tgatcaccca caacggcacc
     2161 gccatcaaga gggccacctt catgagctac aacaccatca tcagcaacag cctgagcttc
     2221 gacatcgtga acaagagcct gcagttcaag tacaagaccc agaaggccac catcctggag
     2281 gccagcctga agaagctgat ccccgcctgg gagttcacca tcatccctta caacggccag
     2341 aagcaccaga gcgacatcac cgacatcgtg tccagcctgc agctgcagtt cgagagcagc
     2401 gaggaggccg acaagggcaa cagccacagc aagaagatgc tgaaggccct gctgtccgag
     2461 ggcgagagca tctgggagat caccgagaag atcctgaaca gcttcgagta caccagcagg
     2521 ttcaccaaga ccaagaccct gtaccagttc ctgttcctgg ccacattcat caactgcggc
     2581 aggttcagcg acatcaagaa cgtggacccc aagagcttca agctggtgca gaacaagtac
     2641 ctgggcgtga tcattcagtg cctggtgacc gagaccaaga caagcgtgtc caggcacatc
     2701 tactttttca gcgccagagg caggatcgac cccctggtgt acctggacga gttcctgagg
     2761 aacagcgagc ccgtgctgaa gagagtgaac aggaccggca acagcagcag caacaagcag
     2821 gagtaccagc tgctgaagga caacctggtg cgcagctaca acaaggccct gaagaagaac
     2881 gccccctacc ccatcttcgc tatcaagaac ggccctaaga gccacatcgg caggcacctg
     2941 atgaccagct ttctgagcat gaagggcctg accgagctga caaacgtggt gggcaactgg
     3001 agcgacaaga gggcctccgc cgtggccagg accacctaca cccaccagat caccgccatc
     3061 cccgaccact acttcgccct ggtgtccagg tactacgcct acgaccccat cagcaaggag
     3121 atgatcgccc tgaaggacga gaccaacccc atcgaggagt ggcagcacat cgagcagctg
     3181 aagggcagcg ccgagggcag catcagatac cccgcctgga acggcatcat cagccaggag
     3241 gtgctggact acctgagcag ctacatcaac aggcggatct gatgtacaag taaagcggcc
     3301 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     3361 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     3421 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3481 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3541 aaggcgtaaa ttgtaagcgt taatgatcca ctagaggatc cccgggtacc gagctcgaat
     3601 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     3661 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     3721 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     3781 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     3841 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3901 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3961 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4021 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     4081 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4141 tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta taagggattt
     4201 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4261 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     4321 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4381 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4441 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     4501 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     4561 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4621 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4681 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4741 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4801 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4861 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4921 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4981 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5041 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5101 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5161 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5221 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5281 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5341 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5401 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5461 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5521 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5581 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5641 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5701 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5761 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5821 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     5881 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5941 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6001 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6061 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6121 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6181 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6241 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6301 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6361 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     6421 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaag
//
