LOCUS       sequence-409183-        6782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of green fluorescent reporter AcGFP driven by the strong 
            Beta-actin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6782)
  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 6782)
  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-AcGFP-CI
FEATURES             Location/Qualifiers
     source          1..6782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1179..1201
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1263..1272
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1269..1985
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     misc_feature    1986..2051
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2178..2312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2215..2234)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2269..2288
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2326..2342)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2326..2342)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2339..2361)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2350..2366
                     /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(2374..2380,2381..2398,2399..2404))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2419..2440
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2557..2574)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2728..3316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2808..2827)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3487..4278,4279..4347))
                     /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     4110..4129
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4348..4452)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4520..4538
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4576..4598)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4698..4717
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4911..4933
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4925..4942
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4926..4942
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(6743..6782,1..236)
                     /label=chicken beta-actin promoter
ORIGIN
        1 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
       61 gggggsgggg 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 gtgggggggt 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 gcgctaccgg
     1261 tcgccaccat ggtgagcaag ggcgccgagc tgttcaccgg catcgtgccc atcctgatcg
     1321 agctgaatgg cgatgtgaat ggccacaagt tcagcgtgag cggcgagggc gagggcgatg
     1381 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cctgtgccct
     1441 ggcccaccct ggtgaccacc ctgagctacg gcgtgcagtg cttctcacgc taccccgatc
     1501 acatgaagca gcacgacttc ttcaagagcg ccatgcctga gggctacatc caggagcgca
     1561 ccatcttctt cgaggatgac ggcaactaca agtcgcgcgc cgaggtgaag ttcgagggcg
     1621 ataccctggt gaatcgcatc gagctgaccg gcaccgattt caaggaggat ggcaacatcc
     1681 tgggcaataa gatggagtac aactacaacg cccacaatgt gtacatcatg accgacaagg
     1741 ccaagaatgg catcaaggtg aacttcaaga tccgccacaa catcgaggat ggcagcgtgc
     1801 agctggccga ccactaccag cagaataccc ccatcggcga tggccctgtg ctgctgcccg
     1861 ataaccacta cctgtccacc cagagcgccc tgtccaagga ccccaacgag aagcgcgatc
     1921 acatgatcta cttcggcttc gtgaccgccg ccgccatcac ccacggcatg gatgagctgt
     1981 acaagtccgg actcagatct cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg
     2041 gcccgggatc caccggatct agataagtaa tgatcataat cagccatatc acatctgtag
     2101 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     2161 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     2221 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2281 aactcatcaa tgtatcttat catgtctgga tcaattcgta atcatgtcat agctgtttcc
     2341 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     2401 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2461 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2521 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2581 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2641 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2701 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2761 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2821 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2881 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2941 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3001 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3061 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3121 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3181 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3241 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3301 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3361 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3421 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3481 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3541 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3601 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3661 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3721 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3781 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3841 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3901 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3961 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4021 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4081 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4141 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4201 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4261 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4321 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4381 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4441 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     4501 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     4561 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4621 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4681 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     4741 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     4801 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     4861 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     4921 acgttgtaaa acgacggcca gtgccaagtt gggatctttg cattggccca cggctctcag
     4981 gatggggatg ctccccttca gcacccggtt ccccttggaa actgatggtc ctggctctgt
     5041 ggcatggcag tggcactgtg aggagccccc taccagcagc acacagtggg gttggcactg
     5101 ccacgctccg gatgccgcgc tctgatccaa ccccataatc aagggaaccc gaattgcccc
     5161 atcattgccc ccaccacccc catcctgccg ggccctcaca ccccacgctg ccttctggtg
     5221 acattcccca gcccaaaccc acggcttcat ggctaccgcg gggcatttcc cattgccgcc
     5281 ccattatcag ctctgcacac ctcccgctgt acccatgcct cgtggctgcc cttctttgac
     5341 gtataatctt ctaattaata cccggccttg tcaaagtgga gcacaaacgt taattaattc
     5401 cccagcaggc aggtaattaa cagtgtgact ccctttttgc tgcgagtggg gctgatacag
     5461 agagatgtgg cactatggag cccacggggt cctggcactg ggtgcccacg gaggtcccca
     5521 tgtgctgcag tgtcaccgcc tccgaggaga cagtattgtc cctgcggtgt ccctgcagct
     5581 cagctctgtc cacagggcca cctccagttt ggaggggaca caatgcagcc ccgatgcaac
     5641 ccatcctcgc agcatcccag ggacaaagac cccactgcaa gaccgcacac agggctgggt
     5701 cccgctcccc taatatctac agtgcttctt gcatggcccc ttaatcaatg cagttaatca
     5761 gcatgcgctc atgcaccgct ctggagctgc aaagcccctc gcagcgctgc tcaccaacac
     5821 cgcgcaccgc cccggcccag cctgcagcac gcgctgcaaa caggaaagaa acaaaatatt
     5881 gcccaaatgt aggcaaaggc attcggctgc cttgacctcc gccgggccgg gccctgcctg
     5941 actcagctcc ttactcagcg ctcgcttcct ccctccggct gccaccgccg cagcgcacac
     6001 cctgacaaag agtggccctt aacgggctct gaggtgcacc cagcagtgca ctcagcagtc
     6061 caagggccgg cctggaggtt tgcaccgcta cgtgctgaca ttagcattga acttggccct
     6121 gggtagtgct gcaggccggg cggggtgggt gtagagagtg cagcgcgcgt tgcacccggt
     6181 gccccttccc ctcccttgca tcccagcagg ctgcacccca gcaccaggcc cgtgcatgca
     6241 tgctcctggt gttattgcag cctggtgcat gcatgcgtct tgatggtgca gcgctgtgca
     6301 tgcatcctcc ttggtgtgta gcagtctagt gcatgcatac ccctcggtgt tattgctgct
     6361 ctgtgcacgc acgctcattg tatcacttca tcccagtgca tgcactcaca ctggagcgat
     6421 tgctgctcgg tgcacgcaca ctcattgtat cactgcagct cagtggctgc acgcacaccg
     6481 gtgttattgc tgctcggtgc gtgcatgcac atcagtgtcg ctgcagctca gtgcatgcac
     6541 gctcattgcc catcgctatc cctgcctctc ctgctggcgc tccccgggag gtgacttcaa
     6601 ggggaccgca ggaccacctc gggggtgggg ggagggctgc acacgcggac cccgctcccc
     6661 ctccccaaca aagcactgtg gaatcaaaaa ggggggaggg gggatggagg ggcgcgtcac
     6721 acccccgccc cacaccctca cctcgaggtg agccccacgt tctgcttcac tctccccatc
     6781 tc
//
