LOCUS       sequence-391800-        6773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of green fluorescent reporter mNeonGreen driven by the 
            strong Beta-actin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6773)
  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 6773)
  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-mNeonGreen-CI
FEATURES             Location/Qualifiers
     source          1..6773
                     /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..1976
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      1524..1533
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1977..2042
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2169..2303
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2206..2225)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2260..2279
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2317..2333)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2317..2333)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2330..2352)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2341..2357
                     /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(2365..2371,2372..2389,2390..2395))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2410..2431
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2548..2565)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2719..3307)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2799..2818)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3478..4269,4270..4338))
                     /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     4101..4120
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4339..4443)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4511..4529
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4567..4589)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4689..4708
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4902..4924
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4916..4933
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4917..4933
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(6734..6773,1..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 gccgcaccct
     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 ggcgaggagg ataacatggc ctctctccca gcgacacatg
     1321 agttacacat ctttggctcc atcaacggtg tggactttga catggtgggt cagggcaccg
     1381 gcaatccaaa tgatggttat gaggagttaa acctgaagtc caccaagggt gacctccagt
     1441 tctccccctg gattctggtc cctcatatcg ggtatggctt ccatcagtac ctgccctacc
     1501 ctgacgggat gtcgcctttc caggccgcca tggtagatgg ctccggctac caagtccatc
     1561 gcacaatgca gtttgaagat ggtgcctccc ttactgttaa ctaccgctac acctacgagg
     1621 gaagccacat caaaggagag gcccaggtga aggggactgg tttccctgct gacggtcctg
     1681 tgatgaccaa ctcgctgacc gctgcggact ggtgcaggtc gaagaagact taccccaacg
     1741 acaaaaccat catcagtacc tttaagtgga gttacaccac tggaaatggc aagcgctacc
     1801 ggagcactgc gcggaccacc tacacctttg ccaagccaat ggcggctaac tatctgaaga
     1861 accagccgat gtacgtgttc cgtaagacgg agctcaagca ctccaagacc gagctcaact
     1921 tcaaggagtg gcaaaaggcc tttaccgatg tgatgggcat ggacgagctg tacaagtccg
     1981 gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat
     2041 ccaccggatc tagataagta atgatcataa tcagccatat cacatctgta gaggttttac
     2101 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     2161 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2221 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2281 atgtatctta tcatgtctgg atcaattcgt aatcatgtca tagctgtttc ctgtgtgaaa
     2341 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2401 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2461 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2521 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2581 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2641 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2701 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2761 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2821 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2881 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2941 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3001 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3061 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3121 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3181 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3241 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3301 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3361 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3421 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3481 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3541 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3601 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3661 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3721 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3781 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3841 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3901 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3961 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4021 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4081 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4141 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4201 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4261 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4321 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4381 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4441 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     4501 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     4561 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     4621 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     4681 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     4741 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     4801 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     4861 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     4921 aacgacggcc agtgccaagt tgggatcttt gcattggccc acggctctca ggatggggat
     4981 gctccccttc agcacccggt tccccttgga aactgatggt cctggctctg tggcatggca
     5041 gtggcactgt gaggagcccc ctaccagcag cacacagtgg ggttggcact gccacgctcc
     5101 ggatgccgcg ctctgatcca accccataat caagggaacc cgaattgccc catcattgcc
     5161 cccaccaccc ccatcctgcc gggccctcac accccacgct gccttctggt gacattcccc
     5221 agcccaaacc cacggcttca tggctaccgc ggggcatttc ccattgccgc cccattatca
     5281 gctctgcaca cctcccgctg tacccatgcc tcgtggctgc ccttctttga cgtataatct
     5341 tctaattaat acccggcctt gtcaaagtgg agcacaaacg ttaattaatt ccccagcagg
     5401 caggtaatta acagtgtgac tccctttttg ctgcgagtgg ggctgataca gagagatgtg
     5461 gcactatgga gcccacgggg tcctggcact gggtgcccac ggaggtcccc atgtgctgca
     5521 gtgtcaccgc ctccgaggag acagtattgt ccctgcggtg tccctgcagc tcagctctgt
     5581 ccacagggcc acctccagtt tggaggggac acaatgcagc cccgatgcaa cccatcctcg
     5641 cagcatccca gggacaaaga ccccactgca agaccgcaca cagggctggg tcccgctccc
     5701 ctaatatcta cagtgcttct tgcatggccc cttaatcaat gcagttaatc agcatgcgct
     5761 catgcaccgc tctggagctg caaagcccct cgcagcgctg ctcaccaaca ccgcgcaccg
     5821 ccccggccca gcctgcagca cgcgctgcaa acaggaaaga aacaaaatat tgcccaaatg
     5881 taggcaaagg cattcggctg ccttgacctc cgccgggccg ggccctgcct gactcagctc
     5941 cttactcagc gctcgcttcc tccctccggc tgccaccgcc gcagcgcaca ccctgacaaa
     6001 gagtggccct taacgggctc tgaggtgcac ccagcagtgc actcagcagt ccaagggccg
     6061 gcctggaggt ttgcaccgct acgtgctgac attagcattg aacttggccc tgggtagtgc
     6121 tgcaggccgg gcggggtggg tgtagagagt gcagcgcgcg ttgcacccgg tgccccttcc
     6181 cctcccttgc atcccagcag gctgcacccc agcaccaggc ccgtgcatgc atgctcctgg
     6241 tgttattgca gcctggtgca tgcatgcgtc ttgatggtgc agcgctgtgc atgcatcctc
     6301 cttggtgtgt agcagtctag tgcatgcata cccctcggtg ttattgctgc tctgtgcacg
     6361 cacgctcatt gtatcacttc atcccagtgc atgcactcac actggagcga ttgctgctcg
     6421 gtgcacgcac actcattgta tcactgcagc tcagtggctg cacgcacacc ggtgttattg
     6481 ctgctcggtg cgtgcatgca catcagtgtc gctgcagctc agtgcatgca cgctcattgc
     6541 ccatcgctat ccctgcctct cctgctggcg ctccccggga ggtgacttca aggggaccgc
     6601 aggaccacct cgggggtggg gggagggctg cacacgcgga ccccgctccc cctccccaac
     6661 aaagcactgt ggaatcaaaa aggggggagg ggggatggag gggcgcgtca cacccccgcc
     6721 ccacaccctc acctcgaggt gagccccacg ttctgcttca ctctccccat ctc
//
