LOCUS       sequence-262693-        7774 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 7774)
  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 7774)
  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..7774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(129..197,198..989)
                     /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(347..366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1649..1668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1902..1919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2072..2077,2078..2095,2096..2102)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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     2115..2137
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2134..2150
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        2406..2785
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2787..3064
                     /label=chicken beta-actin promoter
     intron          3065..4082
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4006..4028
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4090..4109
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(4142..4175)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    4308..4429
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    4541..4662
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    4774..4895
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(4918..4951)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             4961..5338
                     /codon_start=1
                     /gene="HIST1H2BB"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPSKSAPAPKKGSKKAITKAQKKDGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSSK"
     CDS             join(5357..5359,5360..6073)
                     /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             5357..6070
                     /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(5387..5415)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     6010..6030
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(6131..6151)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6549..6560)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6706..6723)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6712..6936
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7133..7152)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7134..7152)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7159..7176)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7159..7175)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7168..7190)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(7316..7771)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7453..7474)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7665..7684
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagcgcgc aattaagttt aaacaagggc aggatgtgtc aaactgccaa tagagaacta
     2221 cttactcttc aggctgaagc tgatggaaca ggtaacaaag gcaaacacta atcatgatca
     2281 gcaagatgaa gcagaaaggg aacaagggga tattaaatgt gtatagacac gctagagaga
     2341 tggctcagca gttaagagaa ctagctggtc tttcagaggt cctgagatca ataccggtca
     2401 tggtcgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     2461 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     2521 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     2581 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2641 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     2701 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     2761 cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc
     2821 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
     2881 agcgatgggg gcgggggggg ggggggggcc ccccccaggc ggggcggggc ggggcgaggg
     2941 gcggggcggg gcgaggcgga aaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
     3001 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
     3061 ggcgggagtc gttgcgcgct gccttccccc cgtgccccgc tccgccgccg cctcgcgccg
     3121 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
     3181 tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg
     3241 tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc
     3301 gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga
     3361 gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc
     3421 cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg
     3481 tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct
     3541 gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg
     3601 gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg
     3661 ggcggggccg cctcgggccg gggagggctc gggggaaggg gcgcggcggc ccccggagcg
     3721 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
     3781 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
     3841 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
     3901 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
     3961 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     4021 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
     4081 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattgctag
     4141 cataacttcg tatagcatac attatacgaa gttatagcca ccgcggtggc ggccgctcta
     4201 gaatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     4261 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     4321 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4381 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ctagaatcat
     4441 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     4501 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     4561 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4621 gcattctagt tgtggtttgt ccaaactcat caatgtatct taactagaat cataatcagc
     4681 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     4741 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     4801 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4861 agttgtggtt tgtccaaact catcaatgta tcttaactag tggatctcat gcctaggata
     4921 acttcgtata gcatacatta tacgaagtta taggcgcgcc atgcctgaac cctctaagtc
     4981 tgctccagcc cctaaaaagg gttctaagaa ggctatcact aaggcgcaga agaaggatgg
     5041 taagaagcgt aagcgcagcc gcaaggagag ctattctatc tatgtgtaca aggttctgaa
     5101 gcaggtccac cccgacaccg gcatctcatc caaggccatg gggatcatga actccttcgt
     5161 caacgacatc ttcgagcgca tcgcgggcga ggcttctcgc ctggctcact acaataagcg
     5221 ctcgaccatc acctccaggg agattcagac ggctgtgcgc ctgctgctgc ctggggagct
     5281 ggctaagcat gctgtgtccg agggcactaa ggcagttacc aagtacacta gctctaagga
     5341 tccaccggtc gccaccatga gcaaaggaga agaacttttc actggagttg tcccaattct
     5401 tgttgaatta gatggtgatg ttaatgggca caaattttct gtccgtggag agggtgaagg
     5461 tgatgctaca aacggaaaac tcacccttaa atttatttgc actactggaa aactacctgt
     5521 tccgtggcca acacttgtca ctactctgac ctatggtgtt caatgctttt cccgttatcc
     5581 ggatcacatg aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga
     5641 acgcactata tctttcaaag atgacgggac ctacaagacg cgtgctgaag tcaagtttga
     5701 aggtgatacc cttgttaatc gtatcgagtt aaagggtatt gattttaaag aagatggaaa
     5761 cattcttgga cacaaactcg agtacaactt taactcacac aatgtataca tcacggcaga
     5821 caaacaaaag aatggaatca aagctaactt caaaattcgc cacaacgttg aagatggttc
     5881 cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt
     5941 accagacaac cattacctgt cgacacaatc tgtcctttcg aaagatccca acgaaaagcg
     6001 tgaccacatg gtccttcttg agtttgtaac tgctgctggg attacacatg gcatggatga
     6061 gctctacaaa tgattaatta aaacctctgg attacaaaat ttgtgaaaga ttgactggta
     6121 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     6181 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     6241 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     6301 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     6361 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     6421 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     6481 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     6541 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     6601 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     6661 ccccgcagga tccactagag cggccgccta gagctcgctg atcagcctcg actgtgcctt
     6721 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     6781 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     6841 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     6901 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     6961 ggggctcgac tagagcttgc ggaaccctta agcttgtcat tcgaatttag acacccacat
     7021 ggtggctcat gaccatctat ctataaatgg atctgatttt catgtctggc agtgtacaga
     7081 agctaactga agaaaggtgg aagacccaca agagttcaag gtacccaatt cgccctatag
     7141 tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     7201 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     7261 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggaaatt
     7321 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     7381 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     7441 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     7501 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     7561 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     7621 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     7681 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     7741 gccgcgctta atgcgccgct acagggcgcg tcag
//
