LOCUS       sequence-392038-        6584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dre-activated Cre-inactivated expression of turbo (t)RFP reporter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6584)
  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 6584)
  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-tRFP-pA-lox
FEATURES             Location/Qualifiers
     source          1..6584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        6..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     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"
     CDS             2739..3434
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     primer_bind     complement(3450..3467)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3456..3680
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    3714..3747
                     /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(3805..3822)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3805..3821)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3814..3836)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3963..4418
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4050..4069)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4260..4281
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4444..4548
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4549..4617,4618..5409)
                     /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(4767..4786)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5580..6168
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6069..6088
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6322..6339
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6456..6477
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6492..6497,6498..6515,6516..6522)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6530..6546
                     /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     6535..6557
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6554..6570
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6554..6570
                     /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 cgcgccrggs ggggsggggs ggggsgrggg
      541 gsggggsggg gsgrggcggr gaggkgcggc 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 gggrgggctc gggggrgggg 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 gagcgagctg atcaaggaga
     2761 acatgcacat gaagctgtac atggagggca ccgtgaacaa ccaccacttc aagtgcacat
     2821 ccgagggaga aggcaagccc tacgagggca cccagaccat gaagatcaag gtggtcgagg
     2881 gcggccctct ccccttcgcc ttcgacatcc tggctaccag cttcatgtac ggcagcaaag
     2941 ccttcatcaa ccacacccag ggcatccccg acttctttaa gcagtccttc cctgagggct
     3001 tcacatggga gagaatcacc acatacgaag acgggggcgt gctgaccgct acccaggaca
     3061 ccagcttcca gaacggctgc atcatctaca acgtcaagat caacggggtg aacttcccat
     3121 ccaacggccc tgtgatgcag aagaaaacac gcggctggga ggccaacacc gagatgctgt
     3181 accccgctga cggcggcctg agaggccaca gccagatggc cctgaagctc gtgggcgggg
     3241 gctacctgca ctgctccttc aagaccacat acagatccaa gaaacccgct aagaacctca
     3301 agatgcccgg cttccacttc gtggaccaca gactggaaag aatcaaggag gccgacaaag
     3361 agacctacgt cgagcagcac gagatggctg tggccaagta ctgcgacctc cctagcaaac
     3421 tggggcacag atgagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     3481 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3541 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3601 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3661 ggatgcggtg ggctctatgg ggatccccat caagctgatc cggaaccctt aatataactt
     3721 cgtataatgt atgctatacg aagttattag gtccctcgac ctgcagccca agctagatcg
     3781 aatttcgagg ctagagcggc cgccactggc cgtcgtttta caacgtcgtg actgggaaaa
     3841 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     3901 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     3961 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4021 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4081 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4141 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4201 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4261 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4321 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4381 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     4441 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4501 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4561 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4621 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4681 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4741 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4801 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4861 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4921 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4981 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5041 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5101 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5161 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5221 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5281 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5341 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5401 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5461 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5521 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5581 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5641 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5701 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5761 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5821 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5881 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5941 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6001 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6061 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6121 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6181 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6241 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     6301 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     6361 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     6421 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     6481 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     6541 taacaatttc acacaggaaa cagctatgac catgaggcgc gccg
//
