LOCUS       sequence-391916-        6661 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 6661)
  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 6661)
  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-DIO-inv[roxSTOProx-ZsGreen-bGH]
FEATURES             Location/Qualifiers
     source          1..6661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        6..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1605..1627
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     protein_bind    complement(1661..1694)
                     /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(1745..1778)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1837..2061
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2050..2067
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2086..2781)
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     regulatory      2778..2787
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2795..2826
                     /label=rox
                     /bound_moiety="Dre recombinase"
     polyA_signal    2855..2989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3109..3243
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3363..3497
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    3664..3695
                     /label=rox
                     /bound_moiety="Dre recombinase"
     protein_bind    3741..3774
                     /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    3825..3858
                     /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(3882..3899)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3882..3898)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3891..3913)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4040..4495
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4127..4146)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4337..4358
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4521..4625
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4626..4694,4695..5486)
                     /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(4844..4863)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5657..6245
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6146..6165
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6399..6416
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6533..6554
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6569..6574,6575..6592,6593..6599)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6607..6623
                     /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     6612..6634
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6631..6647
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6631..6647
                     /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 gsgrggsggr 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 tgtgtgcgkg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct
     1141 gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg
     1201 gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg
     1261 ggcggggccg cctcgggccg gggagggctc gggggagggg 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 ggctctagac caggcagact cgactccgga ataacttcgt ataggatact
     1681 ttatacgaag ttatgcagaa tggtagctgg attgtagctg ctattagcaa tatgaaacct
     1741 cttaataact tcgtatagca tacattatac gaagttatgg cgcgttactt gtacagctag
     1801 agcaactagc tagcgaggtt ctttccgcct cagaagccat agagcccacc gcatccccag
     1861 catgcctgct attgtcttcc caatcctccc ccttgctgtc ctgccccacc ccacccccca
     1921 gaatagaatg acacctactc agacaatgcg atgcaatttc ctcattttat taggaaagga
     1981 cagtgggagt ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa acaacagatg
     2041 gctggcaact agaaggcaca gtcgaggctg atcagcgagc tcggttcagg gcaaggcgga
     2101 gccggaggcg atggcgtgct cggtcaggtg ccacttctgg ttcttggcgt cgctgcggtc
     2161 ctcgcgggtc agcttgtgct ggatgaagtg ccagtcgggc atcttgcggg gcacggactt
     2221 ggccttgtac acggtgtcga actggcagcg caagcggcca ccgtccttca gcagcaggta
     2281 catgctcacg tcgcccttca agatgccctg cttgggcacg gggatgatct tctcgcagga
     2341 gggctcccag ttgtcggtca tcttcttcat cacggggccg tcggcgggga agttcacgcc
     2401 gtagaacttg gactcgtggt acatgcagtt ctcctccacg ctcacggtga tgtcggcgtt
     2461 gcagatgcac acggcgccgt cctcgaacag gaaggagcgg tcccaggtgt agccggcggg
     2521 gcaggagttc ttgaagtagt cgacgatgtc ctgggggtac tcggtgaaca cgcggttgcc
     2581 gtacatgaag gcggcggaca agatgtcctc ggcgaagggc aaggggccgc cctccaccac
     2641 gcacaggttg atggcctgct tgcccttgaa ggggtagccg atgccctcgc cggtgatcac
     2701 gaacttgtgg ccgtccacgc agccctccat gcggtacttc atggtcatct ccttggtcag
     2761 gccgtgcttg gactgggcca tggtggcggc tgcctaactt taaataatgc caattattta
     2821 aagttaatta agggttccgg atcagcttga tggggatcca gacatgataa gatacattga
     2881 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     2941 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3001 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     3061 aaacctctac aaatgtggta tggctgatta tgatcctcta gagtcgcaga tccagacatg
     3121 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     3181 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     3241 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     3301 ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatcc tctagagtcg
     3361 cagatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     3421 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     3481 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     3541 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg
     3601 atcctctaga gtcgcagatc ctctagagtc gcagatctgc aagctttcat ttattcatcg
     3661 cgataacttt aaataatgcc aattatttaa agttaggtgg cgggtcgacc ccctacgcgt
     3721 cacgcctatg gtggcctagc ataacttcgt ataaagtatc ctatacgaag ttatttgcct
     3781 taacccagaa attatcactg ttattcttta gaatggtgca aagaataact tcgtataatg
     3841 tatgctatac gaagttatga attcgatatc aagcggccgc cactggccgt cgttttacaa
     3901 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     3961 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     4021 agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4081 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4141 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4201 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4261 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4321 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4381 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4441 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     4501 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4561 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4621 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4681 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4741 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4801 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4861 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4921 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4981 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5041 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5101 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5161 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5221 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5281 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5341 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5401 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5461 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5521 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5581 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5641 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5701 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5761 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5821 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5881 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5941 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6001 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6061 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6121 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6181 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6241 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6301 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     6361 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     6421 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     6481 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     6541 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     6601 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gaggcgcgcc
     6661 g
//
