LOCUS       sequence-391809-        6764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of red fluorescent reporter mOrange driven by the strong 
            Beta-actin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6764)
  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 6764)
  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: pBactin-mOrange2-CI
FEATURES             Location/Qualifiers
     source          1..6764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1179..1201
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1254..1263
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1260..1967
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     misc_feature    1977..2033
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2160..2294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2197..2216)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2251..2270
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2308..2324)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2308..2324)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2321..2343)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2332..2348
                     /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)."
     promoter        complement(join(2356..2362,2363..2380,2381..2386))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2401..2422
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2539..2556)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2710..3298)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2790..2809)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3469..4260,4261..4329))
                     /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     4092..4111
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4330..4434)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4502..4520
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4558..4580)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4680..4699
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4893..4915
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4907..4924
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4908..4924
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(6725..6764,1..236)
                     /label=chicken beta-actin promoter
ORIGIN
        1 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
       61 gggggcgggg gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg
      121 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
      181 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
      241 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
      301 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
      361 gctgtaatta gcgcttggtt taatgacggc tcgtttcttt tctgtggctg cgtgaaagcc
      421 ttaaagggct ccgggagggc cctttgtgcg ggggggagcg gctcgggggg tgcgtgcgtg
      481 tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc
      541 gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc
      601 ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc
      661 gkgggggggt gagcaggggg tgtgggcgcg gcggtcgggc tgtaaccccc ccctgcaccc
      721 ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgtg cggggcgtgg
      781 cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg
      841 gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg
      901 ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca
      961 gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc
     1021 ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg
     1081 ccttcgtgcg tcgccgcgcc gccgtcccct tctccatctc cagcctcggg gctgtccgca
     1141 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
     1201 cggcggggtt tatatcttcc cttctctgtt cctccgcagc ccccaagcta gcagccacca
     1261 tggtgagcaa gggcgaggag aataacatgg ccatcatcaa ggagttcatg cgcttcaagg
     1321 tgcgcatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     1381 gcccctacga gggctttcag accgctaagc tgaaggtgac caagggtggc cccctgccct
     1441 tcgcctggga catcctgtcc cctcatttca cctacggctc caaggcctac gtgaagcacc
     1501 ccgccgacat ccccgactac ttcaagctgt ccttccccga gggcttcaag tgggagcgcg
     1561 tgatgaacta cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
     1621 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtga
     1681 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggtg
     1741 ccctgaaggg caagatcaag atgaggctga agctgaagga cggcggccac tacacctccg
     1801 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacatcgtcg
     1861 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     1921 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtca ggactcagat
     1981 ctcgagctca agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccaccggat
     2041 ctagataagt aatgatcata atcagccata tcacatctgt agaggtttta cttgctttaa
     2101 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     2161 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     2221 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     2281 atcatgtctg gatcaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc
     2341 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     2401 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2461 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2521 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2581 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2641 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2701 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2761 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2821 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2881 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2941 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3001 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3061 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3121 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3181 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3241 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3301 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3361 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3421 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3481 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3541 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3601 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3661 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3721 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3781 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3841 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3901 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3961 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4021 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4081 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4141 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4201 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4261 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4321 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4381 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4441 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     4501 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     4561 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     4621 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     4681 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     4741 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     4801 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     4861 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     4921 cagtgccaag ttgggatctt tgcattggcc cacggctctc aggatgggga tgctcccctt
     4981 cagcacccgg ttccccttgg aaactgatgg tcctggctct gtggcatggc agtggcactg
     5041 tgaggagccc cctaccagca gcacacagtg gggttggcac tgccacgctc cggatgccgc
     5101 gctctgatcc aaccccataa tcaagggaac ccgaattgcc ccatcattgc ccccaccacc
     5161 cccatcctgc cgggccctca caccccacgc tgccttctgg tgacattccc cagcccaaac
     5221 ccacggcttc atggctaccg cggggcattt cccattgccg ccccattatc agctctgcac
     5281 acctcccgct gtacccatgc ctcgtggctg cccttctttg acgtataatc ttctaattaa
     5341 tacccggcct tgtcaaagtg gagcacaaac gttaattaat tccccagcag gcaggtaatt
     5401 aacagtgtga ctcccttttt gctgcgagtg gggctgatac agagagatgt ggcactatgg
     5461 agcccacggg gtcctggcac tgggtgccca cggaggtccc catgtgctgc agtgtcaccg
     5521 cctccgagga gacagtattg tccctgcggt gtccctgcag ctcagctctg tccacagggc
     5581 cacctccagt ttggagggga cacaatgcag ccccgatgca acccatcctc gcagcatccc
     5641 agggacaaag acctcactgc aagaccgcac acagggctgg gtcccgctcc cctaatatct
     5701 acagtgcttc ttgcatggcc ccttaatcaa tgcagttaat cagcatgcgc tcatgcaccg
     5761 ctctggagct gcaaagcccc tcgcagcgct gctcaccaac accgcgcacc gccccggccc
     5821 agcctgcagc acgcgctgca aacaggaaag aaacaaaata ttgcccaaat gtaggcaaag
     5881 gcattcggct gccttgacct ccgccgggcc gggccctgcc tgactcagct ccttactcag
     5941 cgctcgcttc ctccctccgg ctgccaccgc cgcagcgcac accctgacaa agagtggccc
     6001 ttaacgggct ctgaggtgca cccagcagtg cactcagcag tccaagggcc ggcctggagg
     6061 tttgcaccgc tacgtgctga cattagcatt gaacttggcc ctgggtagtg ctgcaggccg
     6121 ggcggggtgg gtgtagagag tgcagcgcgc gttgcacccg gtgccccttc ccctcccttg
     6181 catcccagca ggctgcaccc cagcaccagg cccgtgcatg catgctcctg gtgttattgc
     6241 agcctggtgc atgcatgcgt cttgatggtg cagcgctgtg catgcatcct ccttggtgtg
     6301 tagcagtcta gtgcatgcat acccctcggt gttattgctg ctctgtgcac gcacgctcat
     6361 tgtatcactt catcccagtg catgcactca cactggagcg attgctgctc ggtgcacgca
     6421 cactcattgt atcactgcag ctcagtggct gcacgcacac cggtgttatt gctgctcggt
     6481 gcgtgcatgc acatcagtgt cgctgcagct cagtgcatgc acgctcattg cccatcgcta
     6541 tccctgcctc tcctgctggc gctccccggg aggtgacttc aaggggaccg caggaccacc
     6601 tcgggggtgg ggggagggct gcacacgcgg accccgctcc ccctccccaa caaagcactg
     6661 tggaatcaaa aaggggggag gggggatgga ggggcgcgtc acacccccgc cccacaccct
     6721 cacctcgagg tgagccccac gttctgcttc actctcccca tctc
//
