LOCUS       sequence-442626-        6629 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dre-activated Cre-inactivated expression of lyn-mScarlet reporter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6629)
  AUTHORS   Vinopal S, Dupraz S, Alfadil E, Pietralla T, Bendre S, Stiess M, 
            Falk S, Camargo Ortega G, Maghelli N, Tolic IM, Smejkal J, Gotz M, 
            Bradke F
  TITLE     Centrosomal microtubule nucleation regulates radial migration of 
            projection neurons independently of polarization in the developing 
            brain.
  JOURNAL   Neuron. 2023 Apr 19;111(8):1241-1263.e16. doi: 
            10.1016/j.neuron.2023.01.020. Epub 2023 Feb 15.
  PUBMED    36796357
REFERENCE   2  (bases 1 to 6629)
  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.neuron.2023.01.020"; journalName: "Neuron"; date: 
            "2023-04-19- 19"; volume: "111"; issue: "8"; pages: "1241-1263.e16"
COMMENT     Sequence Label: pCAG-lox-roxSTOProx-lyn-mScarlet-pA-lox
FEATURES             Location/Qualifiers
     source          1..6629
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        6..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..664
                     /label=chicken beta-actin promoter
     intron          665..1681
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1605..1627
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1694..1713
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1694..1712
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1741..1774
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1816..1847
                     /label=rox
                     /bound_moiety="Dre recombinase"
     polyA_signal    2014..2148
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2268..2402
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2522..2656
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    2685..2716
                     /label=rox
                     /bound_moiety="Dre recombinase"
     regulatory      2733..2742
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2784..3476
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELYK"
     primer_bind     complement(2914..2932)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3182..3201
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3495..3512)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3501..3725
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    3759..3792
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3850..3867)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3850..3866)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3859..3881)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(4007..4462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4144..4165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4356..4375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4489..4593
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4594..4662,4663..5454)
                     /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(4812..4831)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5625..6213
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6114..6133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6367..6384
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6501..6522
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6537..6542,6543..6560,6561..6567)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6575..6591
                     /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     6580..6602
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6599..6615
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6599..6615
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gattcgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc
      421 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
      481 agcgatgggg gcgggggggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg
      541 gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
      601 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
      661 ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
      721 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
      781 tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg
      841 tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc
      901 gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga
      961 gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc
     1021 cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg
     1081 tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct
     1141 gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg
     1201 gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg
     1261 ggcggggccg cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc
     1321 cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag
     1381 ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg
     1441 caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg
     1501 ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg
     1561 tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg
     1621 tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca
     1681 gatccttaat taataatacg actcactata gggggtcgac ccgccaccaa tcatttaaat
     1741 ataacttcgt ataatgtatg ctatacgaag ttattcgcga tgaataaatg aaagcttggg
     1801 ggtcgacccg ccacctaact ttaaataatt ggcattattt aaagttatcg cgatgaataa
     1861 atgaaagctt gcagatctgc gactctagag gatctgcgac tctagaggat cataatcagc
     1921 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     1981 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     2041 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     2101 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg cgactctaga
     2161 ggatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2221 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2281 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2341 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     2401 tctgcgactc tagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2461 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2521 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2581 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2641 ttatcatgtc tggatcccca tcaagctgat ccggaaccct taattaactt taaataattg
     2701 gcattattta aagttaggca gccgccctgc aggccaccat gggatgtatt aaatcaaaaa
     2761 ggaaagacgg ggcccggtac ctggtgagca agggcgaggc agtgatcaag gagttcatgc
     2821 ggttcaaggt gcacatggag ggctccatga acggccacga gttcgagatc gagggcgagg
     2881 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     2941 ccctgccctt ctcctgggac atcctgtccc ctcagttcat gtacggctcc agggccttca
     3001 ccaagcaccc cgccgacatc cccgactact ataagcagtc cttccccgag ggcttcaagt
     3061 gggagcgcgt gatgaacttc gaggacggcg gcgccgtgac cgtgacccag gacacctccc
     3121 tggaggacgg caccctgatc tacaaggtga agctccgcgg caccaacttc cctcctgacg
     3181 gccccgtaat gcagaagaag acaatgggct gggaagcgtc caccgagcgg ttgtaccccg
     3241 aggacggcgt gctgaagggc gacattaaga tggccctgcg cctgaaggac ggcggccgct
     3301 acctggcgga cttcaagacc acctacaagg ccaagaagcc cgtgcagatg cccggcgcct
     3361 acaacgtcga ccgcaagttg gacatcacct cccacaacga ggactacacc gtggtggaac
     3421 agtacgaacg ctccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaag
     3481 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3541 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3601 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3661 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     3721 tatggggatc cccatcaagc tgatccggaa cccttaatat aacttcgtat aatgtatgct
     3781 atacgaagtt attaggtccc tcgacctgca gcccaagcta gatcgaattt cgaggctaga
     3841 gcggccgcca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     3901 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     3961 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggaaat tgtaagcgtt
     4021 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4081 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4141 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     4201 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     4261 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     4321 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     4381 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4441 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4501 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4561 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4621 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4681 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4741 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4801 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4861 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4921 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4981 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5041 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5101 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5161 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5221 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5281 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5341 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5401 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5461 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5521 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5581 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5641 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5701 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5761 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5821 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5881 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5941 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6001 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6061 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6121 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6181 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6241 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6301 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6361 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     6421 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     6481 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     6541 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     6601 ggaaacagct atgaccatga ggcgcgccg
//
