LOCUS       sequence-274428-        7773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  H2B-sfGFP cre reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7773)
  AUTHORS   Nam HS, Capecchi MR
  TITLE     Lrig1 expression prospectively identifies stem cells in the 
            ventricular-subventricular zone that are neurogenic throughout adult
            life.
  JOURNAL   Neural Dev. 2020 Mar 17;15(1):3. doi: 10.1186/s13064-020-00139-5.
  PUBMED    32183906
REFERENCE   2  (bases 1 to 7773)
  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: "Neural 
            Dev."; date: "2020-03-17"; volume: "15(1):3. doi"; pages: " 
            10.1186/s13064-020-00139-5"
COMMENT     Sequence Label: CAG-loxP-3xpA-loxP-H2B-sfGFP
FEATURES             Location/Qualifiers
     source          1..7773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        167..546
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        548..825
                     /label=chicken beta-actin promoter
     intron          826..1842
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1766..1788
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1850..1869
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1902..1935)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    2068..2189
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2301..2422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2534..2655
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(2678..2711)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             2721..3098
                     /codon_start=1
                     /gene="HIST1H2BB"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPSKSAPAPKKGSKKAITKAQKKDGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSSK"
     CDS             join(3117..3119,3120..3833)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             3117..3830
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3147..3175)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3770..3790
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3891..3911)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4309..4320)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4465..4482)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4471..4695
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4892..4911)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4893..4911)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4918..4935)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4918..4934)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4927..4949)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(5076..5531)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5213..5234)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5425..5444
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5558..5662
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5663..5731,5732..6523)
                     /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(5881..5900)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6694..7282
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7183..7202
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7436..7453
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7570..7591
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7606..7611,7612..7629,7630..7636)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7644..7660
                     /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     7649..7671
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7668..7684
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7668..7684
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgatggaac aggtaacaaa ggcaaacact aatcatgatc agcaagatga agcagaaagg
       61 gaacaagggg atattaaatg tgtatagaca cgctagagag atggctcagc agttaagaga
      121 actagctggt ctttcagagg tcctgagatc aataccggtc atggtcgaca ttgattattg
      181 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      241 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      301 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      361 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      421 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      481 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      541 accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca
      601 cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg
      661 gggggggggc gcgcgccggg gggggggggg gggggggggg gggggggsgg ggsgrggcgg
      721 rgaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg
      781 cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc
      841 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
      901 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
      961 cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc
     1021 cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg
     1081 gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg
     1141 gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg
     1201 tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga
     1261 gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt
     1321 gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc
     1381 gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc
     1441 ggggrgggct cgggggrggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg
     1501 gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg
     1561 tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc
     1621 ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg
     1681 ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc
     1741 cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga
     1801 gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg
     1861 gttattgtgc tgtctcatca ttttggcaaa gaattgctag cataacttcg tatagcatac
     1921 attatacgaa gttatagcca ccgcggtggc ggccgctcta gaatcataat cagccatacc
     1981 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     2041 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     2101 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2161 ggtttgtcca aactcatcaa tgtatcttaa ctagaatcat aatcagccat accacatttg
     2221 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     2281 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     2341 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     2401 ccaaactcat caatgtatct taactagaat cataatcagc cataccacat ttgtagaggt
     2461 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     2521 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2581 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     2641 catcaatgta tcttaactag tggatctcat gcctaggata acttcgtata gcatacatta
     2701 tacgaagtta taggcgcgcc atgcctgaac cctctaagtc tgctccagcc cctaaaaagg
     2761 gttctaagaa ggctatcact aaggcgcaga agaaggatgg taagaagcgt aagcgcagcc
     2821 gcaaggagag ctattctatc tatgtgtaca aggttctgaa gcaggtccac cccgacaccg
     2881 gcatctcatc caaggccatg gggatcatga actccttcgt caacgacatc ttcgagcgca
     2941 tcgcgggcga ggcttctcgc ctggctcact acaataagcg ctcgaccatc acctccaggg
     3001 agattcagac ggctgtgcgc ctgctgctgc ctggggagct ggctaagcat gctgtgtccg
     3061 agggcactaa ggcagttacc aagtacacta gctctaagga tccaccggtc gccaccatga
     3121 gcaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta gatggtgatg
     3181 ttaatgggca caaattttct gtccgtggag agggtgaagg tgatgctaca aacggaaaac
     3241 tcacccttaa atttatttgc actactggaa aactacctgt tccgtggcca acacttgtca
     3301 ctactctgac ctatggtgtt caatgctttt cccgttatcc ggatcacatg aaacggcatg
     3361 actttttcaa gagtgccatg cccgaaggtt atgtacagga acgcactata tctttcaaag
     3421 atgacgggac ctacaagacg cgtgctgaag tcaagtttga aggtgatacc cttgttaatc
     3481 gtatcgagtt aaagggtatt gattttaaag aagatggaaa cattcttgga cacaaactcg
     3541 agtacaactt taactcacac aatgtataca tcacggcaga caaacaaaag aatggaatca
     3601 aagctaactt caaaattcgc cacaacgttg aagatggttc cgttcaacta gcagaccatt
     3661 atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac cattacctgt
     3721 cgacacaatc tgtcctttcg aaagatccca acgaaaagcg tgaccacatg gtccttcttg
     3781 agtttgtaac tgctgctggg attacacatg gcatggatga gctctacaaa tgattaatta
     3841 aaacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     3901 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     3961 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     4021 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     4081 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     4141 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     4201 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     4261 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     4321 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     4381 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcagga tccactagag
     4441 cggccgccta gagctcgcta tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     4501 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     4561 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     4621 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     4681 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctcgact agagcttgcg
     4741 gaacccttaa gcttgtcatt cgaatttaga cacccacatg gtggctcatg accatctatc
     4801 tataaatgga tctgattttc atgtctggca gtgtacagaa gctaactgaa gaaaggtgga
     4861 agacccacaa gagttcaagg tacccaattc gccctatagt gagtcgtatt acaattcact
     4921 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     4981 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     5041 ttcccaacag ttgcgcagcc tgaatggcga atggcaaatt gtaagcgtta atattttgtt
     5101 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     5161 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     5221 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     5281 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     5341 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     5401 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     5461 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     5521 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5581 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5641 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5701 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5761 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5821 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5881 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5941 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     6001 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6061 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6121 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6181 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     6241 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6301 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6361 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6421 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6481 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6541 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6601 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6661 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6721 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6781 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6841 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6901 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6961 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7021 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7081 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7141 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7201 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7261 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7321 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     7381 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     7441 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     7501 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     7561 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     7621 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     7681 tgaccatgat tacgccaagc gcgcaattaa gtttaaacaa gggcaggatg tgtcaaactg
     7741 ccaatagaga actacttact cttcaggctg aag
//
