LOCUS       sequence-153724-        6815 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Flp recombinase dependent on GFP (Flp-DOG) for expression in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6815)
  AUTHORS   Tang JC, Drokhlyansky E, Etemad B, Rudolph S, Guo B, Wang S, Ellis 
            EG, Li JZ, Cepko CL
  TITLE     Detection and manipulation of live antigen-expressing cells using 
            conditionally stable nanobodies.
  JOURNAL   Elife. 2016 May 20;5. pii: e15312. doi: 10.7554/eLife.15312.
  PUBMED    27205882
REFERENCE   2  (bases 1 to 6815)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2016-05-20"; volume: "5. pii: e15312. doi"; pages: " 
            10.7554/eLife.15312"
COMMENT     Sequence Label: pCAG-Flp-DOG
FEATURES             Location/Qualifiers
     source          1..6815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1762..1771
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3839..3858)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3904..3959
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3958..3977)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4318..4334)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4318..4334)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4331..4353)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4342..4358
                     /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(4366..4372,4373..4390,4391..4396))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4411..4432
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4491..4686
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4537..4672
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4692..4826
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4729..4748)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4783..4802
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4894..4911)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5065..5653)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5145..5164)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5824..6615,6616..6684))
                     /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     6447..6466
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6685..6789)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attctgcagt cgacggtacc
     1741 gcgggcccgg gatccaccgg tgccaccatg gccgacgtgc agctcgtgga atccggagga
     1801 gcactggtcc agcctggggg atcactgaga ctgtcttgtg tcgcttctgg ctttcctgtg
     1861 aatcggtact ctatgagatg gtacaggcag gcacctggaa aagagcgaga atgggtcgcc
     1921 ggaatgtcat ctgctggcga taggagttcc tacgtggata gcgtgaaagg acggttcacc
     1981 attagaaggg acgatgctag aaacaccgtg tacctccaga tgaactccct gaaacctgag
     2041 gacactgctg tgtactacta caacgtcaat gtgggctttg aatactgggg acatggcaca
     2101 caagtgactg ttagttttgc atgcatggct gacgtccaac ttgttgagag cggtggagca
     2161 ctggtccagc ctgggggatc actgagactg tcttgtgtcg cttctggctt tcctgtgaat
     2221 cggtactcta tgagatggta caggcaggca cctggaaaag agcgagaatg ggtcgccgga
     2281 atgtcatctg ctggcgatag gagttcctac gtggatagcg tgaaaggacg gttcaccatt
     2341 agaagggacg atgctagaaa caccgtgtac ctccagatga actccctgaa acctgaggac
     2401 actgctgtgt actactacaa cgtcaatgtg ggctttgaat actggggcca tgggacacag
     2461 gttactgtta gcttcgcatg catgagccag ttcgacatcc tgtgcaagac cccccccaag
     2521 gtgctggtgc ggcagttcgt ggagagattc gagaggccca gcggcgagaa gatcgccagc
     2581 tgtgccgccg agctgaccta cctgtgctgg atgatcaccc acaacggcac cgccatcaag
     2641 agggccacct tcatgagcta caacaccatc atcagcaaca gcctgagctt cgacatcgtg
     2701 aacaagagcc tgcagttcaa gtacaagacc cagaaggcca ccatcctgga ggccagcctg
     2761 aagaagctga tccccgcctg ggagttcacc atcatccctt acaacggcca gaagcaccag
     2821 agcgacatca ccgacatcgt gtccagcctg cagctgcagt tcgagagcag cgaggaggcc
     2881 gacaagggca acagccacag caagaagatg ctgaaggccc tgctgtccga gggcgagagc
     2941 atctgggaga tcaccgagaa gatcctgaac agcttcgagt acaccagcag gttcaccaag
     3001 accaagaccc tgtaccagtt cctgttcctg gccacattca tcaactgcgg caggttcagc
     3061 gacatcaaga acgtggaccc caagagcttc aagctggtgc agaacaagta cctgggcgtg
     3121 atcattcagt gcctggtgac cgagaccaag acaagcgtgt ccaggcacat ctactttttc
     3181 agcgccagag gcaggatcga ccccctggtg tacctggacg agttcctgag gaacagcgag
     3241 cccgtgctga agagagtgaa caggaccggc aacagcagca gcaacaagca ggagtaccag
     3301 ctgctgaagg acaacctggt gcgcagctac aacaaggccc tgaagaagaa cgccccctac
     3361 cccatcttcg ctatcaagaa cggccctaag agccacatcg gcaggcacct gatgaccagc
     3421 tttctgagca tgaagggcct gaccgagctg acaaacgtgg tgggcaactg gagcgacaag
     3481 agggcctccg ccgtggccag gaccacctac acccaccaga tcaccgccat ccccgaccac
     3541 tacttcgccc tggtgtccag gtactacgcc tacgacccca tcagcaagga gatgatcgcc
     3601 ctgaaggacg agaccaaccc catcgaggag tggcagcaca tcgagcagct gaagggcagc
     3661 gccgagggca gcatcagata ccccgcctgg aacggcatca tcagccagga ggtgctggac
     3721 tacctgagca gctacatcaa caggcggatc taggcggccg cactcctcag gtgcaggctg
     3781 cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc
     3841 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     3901 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     3961 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     4021 gtttggcaac atatgccata tgctggctgc catgaacaaa ggtggctata aagaggtcat
     4081 cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga
     4141 ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt
     4201 tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct
     4261 gtccctcttc tcttatgaag atccctcgac ctgcagccca agcttggcgt aatcatggtc
     4321 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4381 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4441 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat
     4501 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4561 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4621 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4681 tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4741 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4801 tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag
     4861 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4921 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4981 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5041 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5101 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5161 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5221 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     5281 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5341 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5401 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5461 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     5521 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5581 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5641 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5701 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5761 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5821 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5881 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5941 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6001 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6061 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6121 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6181 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6241 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6301 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6361 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6421 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6481 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6541 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6601 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6661 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6721 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6781 ggttccgcgc acatttcccc gaaaagtgcc acctg
//
