LOCUS       sequence-177032-        6830 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses aa1-396 of Flp recombinase fused to ABI CID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6830)
  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 6830)
  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: pBW2286_pCAG-PV1-FlpO-N396-L1-ABI-NLS-BGHpA
FEATURES             Location/Qualifiers
     source          1..6830
                     /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"
     CDS             2798..2821
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2822..2842
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2852..3076
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3316..3335)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3381..3436
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3435..3454)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3797..3813)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3797..3813)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3810..3832)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3821..3837
                     /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(3845..3851,3852..3869,3870..3875))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3890..3911
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      3996..4130
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4150..4284
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4187..4206)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4241..4260
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4352..4369)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4523..5111)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4603..4622)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5282..6073,6074..6142))
                     /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     5905..5924
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6143..6247)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6278..6657
                     /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 gccaccatga gccagttcga catcctgtgc aagacccccc ccaaggtgct
      721 ggtgcggcag ttcgtggaga gattcgagag gcccagcggc gagaagatcg ccagctgtgc
      781 cgccgagctg acctacctgt gctggatgat cacccacaac ggcaccgcca tcaagagggc
      841 caccttcatg agctacaaca ccatcatcag caacagcctg agcttcgaca tcgtgaacaa
      901 gagcctgcag ttcaagtaca agacccagaa ggccaccatc ctggaggcca gcctgaagaa
      961 gctgatcccc gcctgggagt tcaccatcat cccttacaac ggccagaagc accagagcga
     1021 catcaccgac atcgtgtcca gcctgcagct gcagttcgag agcagcgagg aggccgacaa
     1081 gggcaacagc cacagcaaga agatgctgaa ggccctgctg tccgagggcg agagcatctg
     1141 ggagatcacc gagaagatcc tgaacagctt cgagtacacc agcaggttca ccaagaccaa
     1201 gaccctgtac cagttcctgt tcctggccac attcatcaac tgcggcaggt tcagcgacat
     1261 caagaacgtg gaccccaaga gcttcaagct ggtgcagaac aagtacctgg gcgtgatcat
     1321 tcagtgcctg gtgaccgaga ccaagacaag cgtgtccagg cacatctact ttttcagcgc
     1381 cagaggcagg atcgaccccc tggtgtacct ggacgagttc ctgaggaaca gcgagcccgt
     1441 gctgaagaga gtgaacagga ccggcaacag cagcagcaac aagcaggagt accagctgct
     1501 gaaggacaac ctggtgcgca gctacaacaa ggccctgaag aagaacgccc cctaccccat
     1561 cttcgctatc aagaacggcc ctaagagcca catcggcagg cacctgatga ccagctttct
     1621 gagcatgaag ggcctgaccg agctgacaaa cgtggtgggc aactggagcg acaagagggc
     1681 ctccgccgtg gccaggacca cctacaccca ccagatcacc gccatccccg accactactt
     1741 cgccctggtg tccaggtact acgcctacga ccccatcagc aaggagatga tcgccctgaa
     1801 ggacgagacc aaccccatcg aggagtggca gcacatcgag cagctgaagg gcagcgccga
     1861 gggctccgga gggtctggct ccggatcaag tggtggcagc ggtacccctt tgtatggttt
     1921 tacttcgatt tgtggaagaa gacctgagat ggaagatgct gtttcgacta taccaagatt
     1981 ccttcaatct tcctctggtt cgatgttaga tggtcggttt gatcctcaat ccgccgctca
     2041 tttcttcggt gtttacgacg gccatggcgg ttctcaggta gcgaactatt gtagagagag
     2101 gatgcatttg gctttggcgg aggagatagc taaggagaaa ccgatgctct gcgatggtga
     2161 tacgtggctg gagaagtgga agaaagctct tttcaactcg ttcctgagag ttgactcgga
     2221 gattgggtca gttgcgccgg agacggttgg gtcaacgtcg gtggttgccg ttgttttccc
     2281 gtctcacatc ttcgtcgcta actgcggtga ctctagagcc gttctttgcc gcggcaaaac
     2341 tgcacttcca ttatccgttg accataaacc ggatagagaa gatgaagctg cgaggattga
     2401 agccgcagga gggaaagtga ttcagtggaa tggagctcgt gttttcggtg ttctcgccat
     2461 gtcgagatcc attggcgata gatacttgaa accatccatc attcctgatc cggaagtgac
     2521 ggctgtgaag agagtaaaag aagatgattg tctgattttg gcgagtgacg gggtttggga
     2581 tgtaatgacg gatgaagaag cgtgtgagat ggcaaggaag cggattctct tgtggcacaa
     2641 gaaaaacgcg gtggctgggg atgcatcgtt gctcgcggat gagcggagaa aggaagggaa
     2701 agatcctgcg gcgatgtccg cggctgagta tttgtcaaag ctggcgatac agagaggaag
     2761 caaagacaac ataagtgtgg tggtggttga tttgaaggat tacaaggacg atgacgataa
     2821 gcccaagaaa aagcggaagg tgtgagaatt cctgtgcctt ctagttgcca gccatctgtt
     2881 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2941 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3001 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3061 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3121 cacgcgccct gtagcgtgta ccgctgggag ttcgtagacg gaaacaaacg cagaatccaa
     3181 gcgcggccgc ccaggataca tagattacca caactccgag cccttccacc gctagcgcac
     3241 tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca
     3301 caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct
     3361 tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     3421 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa
     3481 tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg
     3541 ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca
     3601 tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc
     3661 tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg
     3721 actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa
     3781 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     3841 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     3901 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3961 agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccagttc
     4021 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4081 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4141 caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4201 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4261 aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag
     4321 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4381 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4441 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4501 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4561 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4621 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4681 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4741 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4801 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4861 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4921 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4981 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5041 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5101 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5161 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5221 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5281 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5341 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5401 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5461 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5521 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5581 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5641 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5701 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5761 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5821 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5881 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5941 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6001 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6061 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6121 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6181 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6241 ttccgcgcac atttccccga aaagtgccac ctgggtcgac attgattatt gactagttat
     6301 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     6361 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     6421 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     6481 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     6541 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     6601 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
     6661 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
     6721 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg ggggggggcg
     6781 cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
//
