LOCUS       sequence-337554-        7377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sfGFP cre reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7377)
  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 7377)
  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-sfGFP
FEATURES             Location/Qualifiers
     source          1..7377
                     /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..4081
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4005..4027
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4089..4108
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(4141..4174)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    4307..4428
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    4540..4661
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    4773..4894
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(4917..4950)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             join(4960..4962,4963..5676)
                     /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             4960..5673
                     /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(4990..5018)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5613..5633
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(5734..5754)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6152..6163)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6308..6325)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6314..6538
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6735..6754)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6736..6754)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6761..6778)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6761..6777)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6770..6792)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(6919..7374)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7056..7077)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7268..7287
                     /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 tactgttctt 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 ggggggggcg cgcgccrggs ggggsggggs ggggsgrggg
     2941 gsggggsggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
     3001 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
     3061 ggcgggagtc gctgcgcgct gccttcgccc 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 gggggagggg cgcggcggcc cccggagcgc
     3721 cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag
     3781 ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg
     3841 caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg
     3901 ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg
     3961 tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg
     4021 tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca
     4081 gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattgctagc
     4141 ataacttcgt atagcataca ttatacgaag ttatagccac cgcggtggcg gccgctctag
     4201 aatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     4261 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     4321 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4381 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac tagaatcata
     4441 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     4501 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     4561 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4621 cattctagtt gtggtttgtc caaactcatc aatgtatctt aactagaatc ataatcagcc
     4681 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     4741 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     4801 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4861 gttgtggttt gtccaaactc atcaatgtat cttaactagt ggatctcatg cctaggataa
     4921 cttcgtatag catacattat acgaagttat aggcgcgcca tgagcaaagg agaagaactt
     4981 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     5041 tctgtccgtg gagagggtga aggtgatgct acaaacggaa aactcaccct taaatttatt
     5101 tgcactactg gaaaactacc tgttccgtgg ccaacacttg tcactactct gacctatggt
     5161 gttcaatgct tttcccgtta tccggatcac atgaaacggc atgacttttt caagagtgcc
     5221 atgcccgaag gttatgtaca ggaacgcact atatctttca aagatgacgg gacctacaag
     5281 acgcgtgctg aagtcaagtt tgaaggtgat acccttgtta atcgtatcga gttaaagggt
     5341 attgatttta aagaagatgg aaacattctt ggacacaaac tcgagtacaa ctttaactca
     5401 cacaatgtat acatcacggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt
     5461 cgccacaacg ttgaagatgg ttccgttcaa ctagcagacc attatcaaca aaatactcca
     5521 attggcgatg gccctgtcct tttaccagac aaccattacc tgtcgacaca atctgtcctt
     5581 tcgaaagatc ccaacgaaaa gcgtgaccac atggtccttc ttgagtttgt aactgctgct
     5641 gggattacac atggcatgga tgagctctac aaatgattaa ttaaaacctc tggattacaa
     5701 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     5761 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     5821 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     5881 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     5941 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     6001 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     6061 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     6121 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     6181 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     6241 tcggatctcc ctttgggccg cctccccgca ggatccacta gagcggccgc ctagagctcg
     6301 ctatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     6361 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     6421 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     6481 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     6541 tctgaggcgg aaagaaccag ctggggctcg actagagctt gcggaaccct taagcttgtc
     6601 attcgaattt agacacccac atggtggctc atgaccatct atctataaat ggatctgatt
     6661 ttcatgtctg gcagtgtaca gaagctaact gaagaaaggt ggaagaccca caagagttca
     6721 aggtacccaa ttcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac
     6781 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     6841 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     6901 gcctgaatgg cgaatggcaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     6961 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     7021 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     7081 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     7141 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     7201 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     7261 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     7321 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcag
//
