LOCUS       sequence-391919-        8280 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual Cre/Dre-dependent expression of ZsGreen. Used as Cre-Dre 
            cotransfection reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8280)
  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 8280)
  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-DIO-inv[roxSTOProx-ZsGreen-bGH]
FEATURES             Location/Qualifiers
     source          1..8280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1308..1341)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1392..1425)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1484..1708
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1697..1714
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1733..2428)
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     regulatory      2425..2434
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2442..2473
                     /label=rox
                     /bound_moiety="Dre recombinase"
     polyA_signal    2502..2636
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2756..2890
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3010..3144
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    3311..3342
                     /label=rox
                     /bound_moiety="Dre recombinase"
     protein_bind    3388..3421
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3472..3505
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3676..3810
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3824..3840)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3824..3840)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3837..3859)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3848..3864
                     /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(3872..3878,3879..3896,3897..3902))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3917..3938
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4055..4072)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4226..4814)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4306..4325)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4985..5776,5777..5845))
                     /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     5608..5627
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5846..5950)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6018..6036
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6074..6096)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6196..6215
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6409..6431
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6423..6440
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6424..6440
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(8241..8280,1..236)
                     /label=chicken beta-actin promoter
ORIGIN
        1 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
       61 gggggcgggg gggggggggg cgcgcgccag gcggggcggg gcggggcgrg 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 gcgttcggct
     1261 tctggcgtgt gaccggcggc tctagaccag gcagactcga ctccggaata acttcgtata
     1321 ggatacttta tacgaagtta tgcagaatgg tagctggatt gtagctgcta ttagcaatat
     1381 gaaacctctt aataacttcg tatagcatac attatacgaa gttatggcgc gttacttgta
     1441 cagctagagc aactagctag cgaggttctt tccgcctcag aagccataga gcccaccgca
     1501 tccccagcat gcctgctatt gtcttcccaa tcctccccct tgctgtcctg ccccacccca
     1561 ccccccagaa tagaatgaca cctactcaga caatgcgatg caatttcctc attttattag
     1621 gaaaggacag tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca
     1681 acagatggct ggcaactaga aggcacagtc gaggctgatc agcgagctcg gttcagggca
     1741 aggcggagcc ggaggcgatg gcgtgctcgg tcaggtgcca cttctggttc ttggcgtcgc
     1801 tgcggtcctc gcgggtcagc ttgtgctgga tgaagtgcca gtcgggcatc ttgcggggca
     1861 cggacttggc cttgtacacg gtgtcgaact ggcagcgcaa gcggccaccg tccttcagca
     1921 gcaggtacat gctcacgtcg cccttcaaga tgccctgctt gggcacgggg atgatcttct
     1981 cgcaggaggg ctcccagttg tcggtcatct tcttcatcac ggggccgtcg gcggggaagt
     2041 tcacgccgta gaacttggac tcgtggtaca tgcagttctc ctccacgctc acggtgatgt
     2101 cggcgttgca gatgcacacg gcgccgtcct cgaacaggaa ggagcggtcc caggtgtagc
     2161 cggcggggca ggagttcttg aagtagtcga cgatgtcctg ggggtactcg gtgaacacgc
     2221 ggttgccgta catgaaggcg gcggacaaga tgtcctcggc gaagggcaag gggccgccct
     2281 ccaccacgca caggttgatg gcctgcttgc ccttgaaggg gtagccgatg ccctcgccgg
     2341 tgatcacgaa cttgtggccg tccacgcagc cctccatgcg gtacttcatg gtcatctcct
     2401 tggtcaggcc gtgcttggac tgggccatgg tggcggctgc ctaactttaa ataatgccaa
     2461 ttatttaaag ttaattaagg gttccggatc agcttgatgg ggatccagac atgataagat
     2521 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     2581 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     2641 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     2701 gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tcctctagag tcgcagatcc
     2761 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2821 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2881 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     2941 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcctct
     3001 agagtcgcag atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     3061 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     3121 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     3181 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
     3241 gattatgatc ctctagagtc gcagatcctc tagagtcgca gatctgcaag ctttcattta
     3301 ttcatcgcga taactttaaa taatgccaat tatttaaagt taggtggcgg gtcgaccccc
     3361 tacgcgtcac gcctatggtg gcctagcata acttcgtata aagtatccta tacgaagtta
     3421 tttgccttaa cccagaaatt atcactgtta ttctttagaa tggtgcaaag aataacttcg
     3481 tataatgtat gctatacgaa gttatgaatt cgatatcaag cggtcgacgg taccgcgggc
     3541 ccgggatcca ccggatctag ataagtaatg atcataatca gccatatcac atctgtagag
     3601 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     3661 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3721 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3781 ctcatcaatg tatcttatca tgtctggatc aattcgtaat catgtcatag ctgtttcctg
     3841 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3901 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3961 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4021 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4081 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4141 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4201 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4261 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4321 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4381 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4441 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4501 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4561 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4621 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4681 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4741 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4801 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4861 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4921 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4981 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5041 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5101 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5161 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5221 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5281 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5341 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5401 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5461 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5521 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5581 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5641 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5701 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5761 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5821 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5881 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5941 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     6001 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     6061 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     6121 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     6181 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     6241 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     6301 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     6361 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     6421 gttgtaaaac gacggccagt gccaagttgg gatctttgca ttggcccacg gctctcagga
     6481 tggggatgct ccccttcagc acccggttcc ccttggaaac tgatggtcct ggctctgtgg
     6541 catggcagtg gcactgtgag gagcccccta ccagcagcac acagtggggt tggcactgcc
     6601 acgctccgga tgccgcgctc tgatccaacc ccataatcaa gggaacccga attgccccat
     6661 cattgccccc accaccccca tcctgccggg ccctcacacc ccacgctgcc ttctggtgac
     6721 attccccagc ccaaacccac ggcttcatgg ctaccgcggg gcatttccca ttgccgcccc
     6781 attatcagct ctgcacacct cccgctgtac ccatgcctcg tggctgccct tctttgacgt
     6841 ataatcttct aattaatacc cggccttgtc aaagtggagc acaaacgtta attaattccc
     6901 cagcaggcag gtaattaaca gtgtgactcc ctttttgctg cgagtggggc tgatacagag
     6961 agatgtggca ctatggagcc cacggggtcc tggcactggg tgcccacgga ggtccccatg
     7021 tgctgcagtg tcaccgcctc cgaggagaca gtattgtccc tgcggtgtcc ctgcagctca
     7081 gctctgtcca cagggccacc tccagtttgg aggggacaca atgcagcccc gatgcaaccc
     7141 atcctcgcag catcccaggg acaaagacct cactgcaaga ccgcacacag ggctgggtcc
     7201 cgctccccta atatctacag tgcttcttgc atggcccctt aatcaatgca gttaatcagc
     7261 atgcgctcat gcaccgctct ggagctgcaa agcccctcgc agcgctgctc accaacaccg
     7321 cgcaccgccc cggcccagcc tgcagcacgc gctgcaaaca ggaaagaaac aaaatattgc
     7381 ccaaatgtag gcaaaggcat tcggctgcct tgacctccgc cgggccgggc cctgcctgac
     7441 tcagctcctt actcagcgct cgcttcctcc ctccggctgc caccgccgca gcgcacaccc
     7501 tgacaaagag tggcccttaa cgggctctga ggtgcaccca gcagtgcact cagcagtcca
     7561 agggccggcc tggaggtttg caccgctacg tgctgacatt agcattgaac ttggccctgg
     7621 gtagtgctgc aggccgggcg gggtgggtgt agagagtgca gcgcgcgttg cacccggtgc
     7681 cccttcccct cccttgcatc ccagcaggct gcaccccagc accaggcccg tgcatgcatg
     7741 ctcctggtgt tattgcagcc tggtgcatgc atgcgtcttg atggtgcagc gctgtgcatg
     7801 catcctcctt ggtgtgtagc agtctagtgc atgcataccc ctcggtgtta ttgctgctct
     7861 gtgcacgcac gctcattgta tcacttcatc ccagtgcatg cactcacact ggagcgattg
     7921 ctgctcggtg cacgcacact cattgtatca ctgcagctca gtggctgcac gcacaccggt
     7981 gttattgctg ctcggtgcgt gcatgcacat cagtgtcgct gcagctcagt gcatgcacgc
     8041 tcattgccca tcgctatccc tgcctctcct gctggcgctc cccgggaggt gacttcaagg
     8101 ggaccgcagg accacctcgg gggtgggggg agggctgcac acgcggaccc cgctccccct
     8161 ccccaacaaa gcactgtgga atcaaaaagg ggggaggggg gatggagggg cgcgtcacac
     8221 ccccgcccca caccctcacc tcgaggtgag ccccacgttc tgcttcactc tccccatctc
//
