LOCUS       sequence-417241-        8322 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinase reporter for FGF8 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8322)
  AUTHORS   You Z, Wang L, He H, Wu Z, Zhang X, Xue S, Xu P, Hong Y, Xiong M, 
            Wei W, Chen Y
  TITLE     Mapping of clonal lineages across developmental stages in human 
            neural differentiation.
  JOURNAL   Cell Stem Cell. 2023 Apr 6;30(4):473-487.e9. doi: 
            10.1016/j.stem.2023.02.007. Epub 2023 Mar 17.
  PUBMED    36933556
REFERENCE   2  (bases 1 to 8322)
  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"; doi: 
            "10.1016/j.stem.2023.02.007"; journalName: "Cell Stem Cell"; date: 
            "2023-04-6- 6"; volume: "30"; issue: "4"; pages: "473-487.e9"
COMMENT     Sequence Label: FGF8-P2A-iCre
FEATURES             Location/Qualifiers
     source          1..8322
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(16..444)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(126..147)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     338..357
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        471..575
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(576..644,645..1436)
                     /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(794..813)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1607..2195
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2096..2115
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2349..2366
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2483..2504
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2519..2524,2525..2542,2543..2549)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2557..2573
                     /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     2562..2584
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2581..2597
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2581..2597
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2618..2636
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             3643..3699
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3706..3726
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4820..5296
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5019..5038)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     promoter        5357..5856
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5392..5415)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5789..5808
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             5896..6495
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5896..5915)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     complement(8128..8147)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8129..8147)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8154..8171)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8154..8170)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8163..8185)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
       61 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
      121 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
      181 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
      241 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
      301 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
      361 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
      421 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
      481 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
      541 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
      601 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
      661 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
      721 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
      781 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
      841 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
      901 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
      961 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     1021 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     1081 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     1141 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     1201 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     1261 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     1321 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     1381 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     1441 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1501 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1561 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1621 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1681 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1741 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1801 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1861 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1921 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1981 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2041 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2101 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2161 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2221 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2281 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     2341 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     2401 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     2461 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     2521 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     2581 caggaaacag ctatgaccat gattacgcca agctcgaaat taaccctcac taaagggaac
     2641 aaaagctggt accagtgagg gagaaggaag ctgtttgaga gatgaaaact ataaaaggcg
     2701 aaggaggaag aatgaaggct agggaagggg ctggggggtg gtggttggta aattccatat
     2761 atctttttaa caaatcgcca aattgctttg ctattatgtc caattacatg gtaccagcat
     2821 ttggaatgtt cagtaagccg caggcccagc ccctcggccg agctctgaaa tggctccatt
     2881 agtcacggct cctctccagc ctcgagctcc ccacttcctg ggcttctggg gctggggtct
     2941 tagcatcttc tcccaggcct cccctccccc atagggtggc tgccctgggg ccagggaacc
     3001 gaagtcctgg gggggtgaga ggggcaggtg gggagacggg tggccagact ggtgggcagg
     3061 aggccagagc aggccaggct ctgggcccct ctctctgtct ttctgcgttg gggcccagcc
     3121 cctccgtaga caaccatgtg tcactgctgc ctgggaggac aggaagttgc cgggtgggct
     3181 gcgagttgtg agggattaga gagcgggtgc ccaggcaggg ggtggggctg cggctcctgc
     3241 ccacctcgcc atctgctggg gtgcccacct gctgtctggg gccgctcgcc ctctgcctct
     3301 gctggggggg ctctgtaacg tggtgtctgg ctcccctacc tgcagagcaa cggcaaaggc
     3361 aaggactgcg tcttcacgga gattgtgctg gagaacaact acacagcgct gcagaatgcc
     3421 aagtacgagg gctggtacat ggccttcacc cgcaagggcc ggccccgcaa gggctccaag
     3481 acgcggcagc accagcgtga ggtccacttc atgaagcggc tgccccgggg ccaccacacc
     3541 accgagcaga gcctgcgctt cgagttcctc aactacccgc ccttcacgcg cagcctgcgc
     3601 ggcagccaga ggacttgggc ccccgagccc cgaggatccg gcgcaacaaa cttctctctg
     3661 ctgaaacaag ccggagatgt cgaagagaat cctggaccga tggtgcccaa gaagaagagg
     3721 aaagtctcca acctgctgac tgtgcaccaa aacctgcctg ccctccctgt ggatgccacc
     3781 tctgatgaag tcaggaagaa cctgatggac atgttcaggg acaggcaggc cttctctgaa
     3841 cacacctgga agatgctcct gtctgtgtgc agatcctggg ctgcctggtg caagctgaac
     3901 aacaggaaat ggttccctgc tgaacctgag gatgtgaggg actacctcct gtacctgcaa
     3961 gccagaggcc tggctgtgaa gaccatccaa cagcacctgg gccagctcaa catgctgcac
     4021 aggagatctg gcctgcctcg cccttctgac tccaatgctg tgtccctggt gatgaggaga
     4081 atcagaaagg agaatgtgga tgctggggag agagccaagc aggccctggc ctttgaacgc
     4141 actgactttg accaagtcag atccctgatg gagaactctg acagatgcca ggacatcagg
     4201 aacctggcct tcctgggcat tgcctacaac accctgctgc gcattgccga aattgccaga
     4261 atcagagtga aggacatctc ccgcaccgat ggtgggagaa tgctgatcca cattggcagg
     4321 accaagaccc tggtgtccac agctggtgtg gagaaggccc tgtccctggg ggttaccaag
     4381 ctggtggaga gatggatctc tgtgtctggt gtggctgatg accccaacaa ctacctgttc
     4441 tgccgggtca gaaagaatgg tgtggctgcc ccttctgcca cctcccaact gtccacccgg
     4501 gccctggaag ggatctttga ggccacccac cgcctgatct atggtgccaa ggatgactct
     4561 gggcagagat acctggcctg gtctggccac tctgccagag tgggtgctgc cagggacatg
     4621 gccagggctg gtgtgtccat ccctgaaatc atgcaggctg gtggctggac caatgtgaac
     4681 attgtgatga actacatcag aaacctggac tctgagactg gggccatggt gaggctgctc
     4741 gaggatgggg actgatagga attctcaatt tctgagaact gtcattctcg gaaattgaag
     4801 cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     4861 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     4921 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     4981 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     5041 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     5101 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     5161 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     5221 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     5281 tcccttccct gtcctttagg tctgaagagg agtttacgtc cagccacgcg taggatctcg
     5341 acctcgaaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct
     5401 ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc
     5461 acatccaccg gtaggcgcca accgactccg ttctttggtg gccccttcgc gccaccttct
     5521 actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga
     5581 caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg
     5641 aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct
     5701 cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg
     5761 ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc
     5821 gcgctgttct cctcttcctc atctccgggc ctttcgacct gcatccatct agatctcgag
     5881 cagctgaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     5941 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     6001 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     6061 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     6121 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     6181 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     6241 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     6301 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     6361 gtgcccgcct tcctggagac ctccgcgccc cacaacctcc ccttctacga gcggctcggc
     6421 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     6481 aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga
     6541 ccccatgcat cgatgatatc agatccccgg gatgcagaaa ttgatgatct attaaacaat
     6601 aaagatgtcc actaaaatgg aagtttttcc tgtcatactt tgttaagaag ggtgagaaca
     6661 gagtacctac attttgaatg gaaggattgg agctacgggg gtgggggtgg ggtgggatta
     6721 gataaatgcc tgctctttac tgaaggctct ttactattgc tttatgataa tgtttcatag
     6781 ttggatatca taatttaaac aagcaaaacc aaattaaggg ccagctcatt cctcccactc
     6841 atgatctata gatctataga tctctcgtgg gatcattgtt tttctcttga ttcccacttt
     6901 gtggttctaa gtactgtggt ttccaaatgt gtcagtttca tagcctgaag aacgagatca
     6961 gcagcctctg ttccacatac acttcattct cagtattgtt ttgccaagtt ctaattccat
     7021 cagaagctgg tcgagatccg gaacccttaa ttcaatttct gagaactgtc attctcggaa
     7081 attgaggatc ctgagtctta ggtgctgcct ttaccgaaac acaatgccag accgcagaga
     7141 ggctcatcgg tgagggcacc caaaactcaa gcaagatgag ctgtgcgctg ctctgcaggc
     7201 tggggaggtg ctgggggagc cctgggttcc ggttgttgat attgtttgct gttgggtttt
     7261 tgctgttttt tttttttttt tttttttaaa acaaaagaga ggctctattt ttgtattcca
     7321 cttggctgtg gtgtctgtct tcttaactct cagaaagctc cattagtggc ctagactggg
     7381 attccggctg ggggtttgcg ggggtggggg gctttctcta gcctgtgctg ctgaggcccc
     7441 agtacctcca gggccagttg gctgggcagc cagggactcc actgcacccc caggtggggc
     7501 agggaggaaa ggactgtgac atagggcagt cctcttagaa gtgggtatca gactggtggc
     7561 tattaaatga ttgaaatatt tatttaactt gcatattaaa aatgtgtgct ggagagtgag
     7621 tcctgccggg gtcagcccct ccctccaacc ttgccccagc tggtgggcgg ctgggagacg
     7681 cagatgacca ggtgccagct ctgaccacag cctccctcca gcctaaagac acctgcctgt
     7741 caaccatccc catcactgtc acttgagggg ttttcctgca aggacagaag cagggaaagg
     7801 ggcaagaaga ggctcttagc tagtccttgg agctctcaga tgtgtacctc ctagcacttt
     7861 acagaggtca ttgctaacac ttcccaggcc acctcagggc cagaaataat ggatgtgcta
     7921 gggctagagc tgtaatcatg gatttaatcc tcttaaaaag tgcttctctg agtgcctagg
     7981 tccatgtggg agacaggttg gagattccag aacttgctct ttctgagact caggctccag
     8041 aaaatgaaag aaaagagcag ctgccagggt ccaaggtggg ggcatattgg aggggggcgg
     8101 ccgccaccgc ggtggagctc caattcgccc tatagtgagt cgtattacaa ttcactggcc
     8161 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     8221 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     8281 caacagttgc gcagcctgaa tggcgaatgg aaattgtaag cg
//
