LOCUS       sequence-177168-        5323 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses aa397-423 of Flp recombinase fused to PYL CID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5323)
  AUTHORS   Weinberg BH, Pham NT, Caraballo LD, Lozanoski T, Engel A, Bhatia S, 
            Wong WW
  TITLE     Large-scale design of robust genetic circuits with multiple inputs 
            and outputs for mammalian cells.
  JOURNAL   Nat Biotechnol. 2017 Mar 27. doi: 10.1038/nbt.3805.
  PUBMED    28346402
REFERENCE   2  (bases 1 to 5323)
  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 
            Biotechnol. 2017 Mar 27. doi: 10.1038/nbt.3805."
COMMENT     Sequence Label: pBW2287_pCAG-PV1-PYL-L1-FlpO-397C-NLS-BGHpA
FEATURES             Location/Qualifiers
     source          1..5323
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     466..488
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     550..569
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      671..680
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1217..1243
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1367..1387
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1397..1621
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1861..1880)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1926..1981
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1980..1999)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2342..2358)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2342..2358)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2355..2377)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2366..2382
                     /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(2390..2396,2397..2414,2415..2420))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2435..2456
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2541..2675
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2695..2829
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2732..2751)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2786..2805
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2897..2914)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3068..3656)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3148..3167)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3827..4618,4619..4687))
                     /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     4450..4469
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4688..4792)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4823..5202
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg
       61 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
      121 gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg
      181 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
      241 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
      301 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
      361 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
      421 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
      481 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
      541 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattgatta
      601 attcgagcga aggcgcgccc attactcgca tccattctca ggctgtctcg tctcgtctcg
      661 cacgcgtgcc gccaccatgg cgccaactca agacgagttc acccaactct cccaatcaat
      721 cgccgagttc cacacgtacc aactcggtaa cggccgttgc tcatctctcc tagctcagcg
      781 aatccacgcg ccgccggaaa cagtatggtc cgtggtgaga cgtttcgata ggccacagat
      841 ttacaaacac ttcatcaaaa gctgtaacgt gagtgaagat ttcgagatgc gagtgggatg
      901 cacgcgcgac gtgaacgtga taagtggatt accggcgaat acgtctcgag agagattaga
      961 tctgttggac gatgatcgga gagtgactgg gtttagtata accggtggtg aacataggct
     1021 gaggaattat aaatcggtta cgacggttca tagatttgag aaagaagaag aagaagaaag
     1081 gatctggacc gttgttttgg aatcttatgt tgttgatgta ccggaaggta attcggagga
     1141 agatacgaga ttgtttgctg atacggttat tagattgaat cttcagaaac ttgcttcgat
     1201 cactgaagct atgaactacc catacgatgt tccagattac gcttccggag ggtctggctc
     1261 cggatcaagt ggtggcagcg gtaccagcat cagatacccc gcctggaacg gcatcatcag
     1321 ccaggaggtg ctggactacc tgagcagcta catcaacagg cggatcccca agaaaaagcg
     1381 gaaggtgtga gaattcctgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1441 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     1501 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     1561 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     1621 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     1681 gtgtaccgct gggagttcgt agacggaaac aaacgcagaa tccaagcgcg gccgcccagg
     1741 atacatagat taccacaact ccgagccctt ccaccgctag cgcactcctc aggtgcaggc
     1801 tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga
     1861 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
     1921 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     1981 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
     2041 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg
     2101 tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac
     2161 ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa
     2221 atttttctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat
     2281 agctgtccct cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat
     2341 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     2401 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     2461 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct
     2521 caattagtca gcaaccatag tcccgcccct aactccgccc agttccgccc attctccgcc
     2581 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     2641 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg
     2701 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2761 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     2821 gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2881 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2941 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3001 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3061 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3121 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3181 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3241 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3301 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3361 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3421 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3481 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3541 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3601 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3661 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3721 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3781 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3841 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3901 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3961 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4021 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4081 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4141 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4201 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4261 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4321 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4381 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4441 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4501 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4561 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4621 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4681 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4741 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4801 cccgaaaagt gccacctggg tcgacattga ttattgacta gttattaata gtaatcaatt
     4861 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     4921 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     4981 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     5041 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     5101 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     5161 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
     5221 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
     5281 ttttaattat tttgtgcagc gatgggggcg gggggggggg ggg
//
