LOCUS       sequence-352801-        8103 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes 2xNLS::GFP(flexon), codon optimized for C. elegans. Blocks 
            expression from ubiquitous promoter rps-27p with Flexon. Excision of
            flexon with Cre allows for strong expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8103)
  AUTHORS   Shaffer JM, Greenwald I
  TITLE     Floxed exon (Flexon): A flexibly positioned stop cassette for 
            recombinase-mediated conditional gene expression.
  JOURNAL   Proc Natl Acad Sci U S A. 2022 Jan 18;119(3). pii: 2117451119. doi: 
            10.1073/pnas.2117451119.
  PUBMED    35027456
REFERENCE   2  (bases 1 to 8103)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2022-01-18"; volume: "119(3). pii: 
            2117451119. doi"; pages: " 10.1073/pnas.2117451119"
COMMENT     Sequence Label: pJS145
FEATURES             Location/Qualifiers
     source          1..8103
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     283..299
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     283..299
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    1186..1210
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1213..1233
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(1593..1626)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1718..1751)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             2274..2345
                     /codon_start=1
                     /product="nuclear localization signal from the C. elegans 
                     EGL-13 transcription factor (Lyssenko et al., 2007)"
                     /label=EGL-13 NLS
                     /translation="SRRRKANPTKLSENAKKLAKEVEN"
     protein_bind    complement(2347..2371)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3102..3119)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3102..3118)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3940..4734
                     /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(3994..4013)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4604..4623
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5435..5451
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5436..5452
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6107..6129
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6167..6185)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6253..6357
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6358..6426,6427..7218)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6576..6595)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7389..7977
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7878..7897
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcaccgtacg tctcgaggaa ttctatcagg tgtacaagta tgaaatgtcg tttcgaacat
       61 atagatgtct cgcaaacgta aatatttatc gattgtcata aaactttgac cttgtgaagt
      121 gtcaaccttg actgtcgaac caccatagtt tggcgcgaat tgagcgtcat aattgtttac
      181 tctcagtgca gtcaacatgt cgagtttcgt gccgaataaa gagcaaacgc ggacagtatt
      241 aattttctgt tttcatttga agaaaacagc tgcggccgga tccaggaaac agctatgacc
      301 atgattacgc caagctacgt aatacgactc actagtgggc attcaatcgg tttttccttg
      361 cttgcgccca attctgatgg ttcttccatt tctaaatttt agtttaataa aaaaatttta
      421 aaataatacg agttttgaga ctaactttca ataaaaacca aaaaaaattg cttcaaaaaa
      481 gagttcgcgc cgatttacga taaacacagc gcgctacgac actccgaaaa ccgcgccgca
      541 ccattacatt ctccgcccca ccgcacggtt tgcggaaggc gatgaaattg ctgggcctaa
      601 aattttaacc ctttttttcc cgtttttcaa ctgttttccc gattttttcg atattttttt
      661 caggttttta gttagaaaag tatataaaaa ttgcattatt tgtagttttc gctcgtaatt
      721 tgtgatagaa aacggccaat ttgcctagat ttgtgattct accaagtgga atttttggga
      781 aaatatcaat attccacgat ttttattcaa atcataacgt ttcgaaatta tcagcttaaa
      841 taaaactaaa ataaataata aaatactaaa aaccctgaat ttctcggttc tcggttcagc
      901 ttgagttcgt tctatacacg ttatcgattt ttgtcatttc gtttaatcat gggttttgat
      961 tcattttaaa tatgaaattt gatttattcc tcgtttcgaa gttggtttgt cttgtccggc
     1021 cggtaaacaa atgttattaa aaaaataata aatttcgaaa tttttcgtaa tgttccgcaa
     1081 tcgtgttttt ccgtttatct cgtcttccct tctttcaaat atactattta ttttctcaat
     1141 tcacatattt tcagccagcc ctgctcaaca agtggaataa aaatggcaag tttgtacaaa
     1201 aaagcaggct cgccaaagaa gaagcgtaag gtgtccaagg taagttttct agagaccttt
     1261 actaactaac taggtctcat agattttcag ggagaggagc tcttcaccgg agtcgtccca
     1321 atcctcgtcg agctcgacgg agacgtcaac ggacacaagt tctccgtctc cggagaggga
     1381 gagggagacg ccacctacgg aaagctcacc ctcaagttca tctgcaccac cggaaagctc
     1441 ccagtcccat ggccaaccct cgtcaccacc ttctgctacg gagtccaatg cttctcccgt
     1501 tacccagacc acatgaagcg tcacgacttc ttcaagtccg ccatgccaga gggatacgtc
     1561 caagagcgta ccatcttctt cgtaagttta aaataacttc gtataggata ctttatacga
     1621 agttattttc aggacggtga ttaagataaa tgcaaaatcc tttcaagcat tcccttcttc
     1681 tctatcactc ttctttcttt ttcaaggtaa gtttaaaata acttcgtata ggatacttta
     1741 tacgaagtta ttttcagaag gacgacggaa actacaagac ccgtgccgag gtcaagttcg
     1801 agggagacac cctcgtcaac cgtatcgagc tcaagggaat cgacttcaag gaggacggaa
     1861 acatcctcgg acacaagctc gagtacaact acaactccca caacgtctac atcatggccg
     1921 acaagcaaaa gaacggaatc aaggtcaact tcaaggtaag ttctttagag acctttacta
     1981 actaactagg tctcagtaat tttcagatcc gtcacaacat cgaggacgga tccgtccaac
     2041 tcgccgacca ctaccaacaa aacaccccaa tcggagacgg accagtcctc ctcccagaca
     2101 accactacct ctccacccaa tccgccctct ccaaggtaag ttcataagag acctttacta
     2161 actaactagg tctcagaatt tttcaggacc caaacgagaa gcgtgaccac atggtcctcc
     2221 tcgagttcgt caccgccgcc ggaatcaccc acggaatgga cgagctctac aagtcccgtc
     2281 gtcgtaaggc caacccaacc aagctctccg agaacgccaa gaagctcgcc aaggaggtcg
     2341 agaacaaccc agctttcttg tacaaagtgg gataacttaa ggtccaatta ctcttcaaca
     2401 tccctacatg ctctttctcc ctgtgctccc accccctatt tttgttatta tcaaaaaact
     2461 tctcttaatt tctttgtttt ttagcttctt ttaagccacc tctaacaatg aaattgtgta
     2521 gattcaaaaa tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct ccctcccccc
     2581 attaataata attctatccc aaaatctaca caatgttctg tgtacacttc ttatgttttt
     2641 tacttctgat aaattttttt gaaacatcat agaaaaaacc gcacacaaaa taccttatca
     2701 tatgttacgt ttcagtttat gaccgcaatt tttatttctt cgcacgtctg ggcctctcat
     2761 gacgtcaaat catgctcatc gtgaaaaagt tttggagtat ttttggaatt tttcaatcaa
     2821 gtgaaagttt atgaaattaa ttttcctgct tttgcttttt ggggtttccc ctattgtttg
     2881 tcaagatttc gaggacggcg tttttcttgc taaaatcaca agtattgatg agcacgatgc
     2941 aagaaagatc ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta gaagttgata
     3001 atttgaaagt ggagtagtgt ctatggggtt tttgccttaa atgacagaat acattcccaa
     3061 tataccaaac ataactgttt ttaaagtgag tcgtattacg gactggccgt cgttttacag
     3121 gccggatggg ccatggcacc ttaacatttc taaattttag tttaataaaa aaattttaaa
     3181 ataatacgag ttttgagact aactttcaat aaaaaccaaa aaattgcttc aaaaaagagc
     3241 tcgcgccgat ttacgataaa cacagcgcgc tacgacactc cgaaaaccgc gccgcaccat
     3301 tacattctcc gccccaccgc acggtttgcg gaaggcgatg aaattgctgg gcctaaaatt
     3361 ttaacccttt ttttcccgtt tttcaactgt tttcccgatt ttttcgatat ttttttcagg
     3421 tttttagtta gaaaagtata taaaaattgc attatttgta gttttcgctc gtaatttgtg
     3481 atagaaaacg gccaatttgc ctagatttgt gattctacca agtggaattt ttgggaaaat
     3541 atcaatattc cacgattttt attcaaatca taacgtttcg aaattatcag cttaaataaa
     3601 actaaaataa ataataaaat actaaaaacc ctgaatttct cggttctcgg ttcagcttga
     3661 gttcgttcta tacacgttat cgatttttgt catttcgttt aatcatgggt tttgattcat
     3721 tttaaatatg aaatttgatt tattcctcgt ttcgaagttg gtttgtcttg tccggccggt
     3781 aaacaaatgt tattaaaaaa aataataaat ttcgaaattt ttcgtaatgt tccgcaatcg
     3841 tgtttttccg tttatctcgt cttcccttcc ttcaaatata ctatttattt tctcaattca
     3901 catattttca gccagccctg ctcaacaagt ggaataaaaa tgattgaaca agatggattg
     3961 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4021 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4081 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4141 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4201 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4261 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4321 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4381 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcacca
     4441 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4501 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4561 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4621 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4681 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagtccaa
     4741 ttactcttca acatccctac atgctctttc tccctgtgct cccaccccct atttttgtta
     4801 ttatcaaaaa acttctctta atttctttgt tttttagctt cttttaagcc acctctaaca
     4861 atgaaattgt gtagattcaa aaatagaatt aattcgtaat aaaaagtcga aaaaaattgt
     4921 gctccctccc cccattaata ataattctat cccaaaatct acacaatgtt ctgtgtacac
     4981 ttcttatgtt ttttacttct gataaatttt tttgaaacat catagaaaaa accgcacaca
     5041 aaatacctta tcatatgtta cgtttcagtt tatgaccgca atttttattt cttcgcacgt
     5101 ctgggcctct catgacgtca aatcatgctc atcgtgaaaa agttttggag tatttttgga
     5161 atttttcaat caagtgaaag tttatgaaat taattttcct gcttttgctt tttggggttt
     5221 cccctattgt ttgtcaagat ttcgaggacg gcgtttttct tgctaaaatc acaagtattg
     5281 atgagcacga tgcaagaaag atcggaagaa ggtttgggtt tgaggctcag tggaaggtga
     5341 gtagaagttg ataatttgaa agtggagtag tgtctatggg gtttttgcct taaatgacag
     5401 aatacattcc caatatacca aacataactg tttctcgagg tcgacggtat cgataagctt
     5461 gatccggatc tgcggcttac tcaccagacc tggccccatc cgattaccac ctattcgctt
     5521 cgatgggaca cgcactcgct gagcagcgct tcgattctta cgaaagtgtg aaaaaatggc
     5581 tcgatgaatg gttcgccgca aaagacgatg agttctactg gcgtggaatc cacaaattgc
     5641 ccgagagatg ggaaaaatgt gtagctagcg acggcaaata ctttgaataa atgatttttt
     5701 ctttttccac aaaatttaac gtgttttttg atttaaaaaa aacgacattt catacttgta
     5761 cacctgatat ctagatacct aggtgagctc tggtaccccc ctggcgttac ccaacttaat
     5821 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     5881 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     5941 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtgc aatctgctct
     6001 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     6061 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     6121 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     6181 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     6241 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6301 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6361 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     6421 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6481 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6541 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6601 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6661 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6721 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6781 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6841 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6901 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6961 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     7021 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     7081 ggtatcattg cagcactggg gtcagatggt aagccctccc gtatcgtagt tatctacacg
     7141 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     7201 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     7261 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7321 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7381 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     7441 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7501 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7561 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7621 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7681 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7741 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7801 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7861 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7921 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7981 gccagcaacg cggccttttt acggttcctg gtcttttgct ggccttttgc tcacatgttc
     8041 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     8101 acc
//
