LOCUS       sequence-375583-        7842 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual plasmid labeling membrane target protein of different neuronal 
            population with different fluorophore.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7842)
  AUTHORS   Alfadil E, Bradke F, Dupraz S
  TITLE     In Situ Visualization of Axon Growth and Growth Cone Dynamics in 
            Acute Ex Vivo Embryonic Brain Slice Cultures.
  JOURNAL   J Vis Exp. 2021 Oct 14;(176). doi: 10.3791/63068.
  PUBMED    34723948
REFERENCE   2  (bases 1 to 7842)
  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"; journalName: "J Vis 
            Exp."; date: "2021-10-14"; volume: "(176). doi"; pages: " 
            10.3791/63068"
COMMENT     Sequence Label: pCAG-lox-rox-STOP-rox-lynScarlet-lox-Lyn-NeonG...
FEATURES             Location/Qualifiers
     source          1..7842
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        6..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..664
                     /label=chicken beta-actin promoter
     intron          665..1681
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1605..1627
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     protein_bind    1697..1730
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1772..1803
                     /label=rox
                     /bound_moiety="Dre recombinase"
     polyA_signal    1970..2104
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2224..2358
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    2387..2418
                     /label=rox
                     /bound_moiety="Dre recombinase"
     regulatory      2435..2444
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2616..2634)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2884..2903
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3202..3426
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3462..3481)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3527..3582
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3581..3600)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    3931..3964
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      3984..3993
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4035..4745
                     /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      4290..4299
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4767..4991
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5063..5080)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5063..5079)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5072..5094)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(5220..5675)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5357..5378)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5569..5588
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5702..5806
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5807..5875,5876..6667)
                     /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(6025..6044)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6838..7426
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7327..7346
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7580..7597
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7714..7735
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7750..7755,7756..7773,7774..7780)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7788..7804
                     /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     7793..7815
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7812..7828
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7812..7828
                     /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 gsgrggcgga gaggtgcggc 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 tgtgtgcgtg 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 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca
     1681 gatccttaat ttaaatataa cttcgtataa tgtatgctat acgaagttat tcgcgatgaa
     1741 taaatgaaag cttgggggtc gacccgccac ctaactttaa ataattggca ttatttaaag
     1801 ttatcgcgat gaataaatga aagcttgcag atctgcgact ctagaggatc tgcgactcta
     1861 gaggatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     1921 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     1981 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     2041 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     2101 gatctgcgac tctagaggat cataatcagc cataccacat ttgtagaggt tttacttgct
     2161 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     2221 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2281 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2341 tcttatcatg tctcgatccc catcaagctg atccggaacc cttaattaac tttaaataat
     2401 tggcattatt taaagttagg cagccgccct gcaggccacc atgggatgta ttaaatcaaa
     2461 aaggaaagac ggggcccggt acctggtgag caagggcgag gcagtgatca aggagttcat
     2521 gcggttcaag gtgcacatgg agggctccat gaacggccac gagttcgaga tcgagggcga
     2581 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
     2641 ccccctgccc ttctcctggg acatcctgtc ccctcagttc atgtacggct ccagggcctt
     2701 caccaagcac cccgccgaca tccccgacta ctataagcag tccttccccg agggcttcaa
     2761 gtgggagcgc gtgatgaact tcgaggacgg cggcgccgtg accgtgaccc aggacacctc
     2821 cctggaggac ggcaccctga tctacaaggt gaagctccgc ggcaccaact tccctcctga
     2881 cggccccgta atgcagaaga agacaatggg ctgggaagcg tccaccgagc ggttgtaccc
     2941 cgaggacggc gtgctgaagg gcgacattaa gatggccctg cgcctgaagg acggcggccg
     3001 ctacctggcg gacttcaaga ccacctacaa ggccaagaag cccgtgcaga tgcccggcgc
     3061 ctacaacgtc gaccgcaagt tggacatcac ctcccacaac gaggactaca ccgtggtgga
     3121 acagtacgaa cgctccgagg gccgccactc caccggcggc atggacgagc gtacaagtaa
     3181 gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     3241 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     3301 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3361 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     3421 ctatggggag ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa
     3481 aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt
     3541 attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg
     3601 agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc
     3661 catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag
     3721 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt
     3781 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
     3841 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctcgga
     3901 tccccatcaa gctgatccgg aacccttaat ataacttcgt ataatgtatg ctatacgaag
     3961 ttattaggtc cctcgacctg caggccacca tgggatgtat taaatcaaaa aggaaagacg
     4021 gggcccggta ccatatggtg agcaagggcg aggaggataa catggcctct ctcccagcga
     4081 cacatgagtt acacatcttt ggctccatca acggtgtgga ctttgacatg gtgggtcagg
     4141 gcaccggcaa tccaaatgat ggttatgagg agttaaacct gaagtccacc aagggtgacc
     4201 tccagttctc cccctggatt ctggtccctc atatcgggta tggcttccat cagtacctgc
     4261 cctaccctga cgggatgtcg cctttccagg ccgccatggt agatggctcc ggctaccaag
     4321 tccatcgcac aatgcagttt gaagatggtg cctcccttac tgttaactac cgctacacct
     4381 acgagggaag ccacatcaaa ggagaggccc aggtgaaggg gactggtttc cctgctgacg
     4441 gtcctgtgat gaccaactcg ctgaccgctg cggactggtg caggtcgaag aagacttacc
     4501 ccaacgacaa aaccatcatc agtaccttta agtggagtta caccactgga aatggcaagc
     4561 gctaccggag cactgcgcgg accacctaca cctttgccaa gccaatggcg gctaactatc
     4621 tgaagaacca gccgatgtac gtgttccgta agacggagct caagcactcc aagaccgagc
     4681 tcaacttcaa ggagtggcaa aaggccttta ccgatgtgat gggcatggac gagctgtaca
     4741 agtaagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     4801 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     4861 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     4921 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     4981 gggctctatg gcttctgagg cggaaagaac cagctggggc tcgagatcca ctagttctag
     5041 cctcgaggct agagcggccg ccactggccg tcgttttaca acgtcgtgac tgggaaaacc
     5101 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     5161 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
     5221 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     5281 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     5341 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     5401 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     5461 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     5521 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     5581 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     5641 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     5701 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5761 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5821 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5881 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5941 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     6001 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6061 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     6121 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6181 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     6241 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6301 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6361 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6421 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6481 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6541 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6601 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6661 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6721 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6781 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6841 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6901 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6961 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     7021 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     7081 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7141 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7201 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7261 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7321 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7381 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7441 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     7501 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     7561 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     7621 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     7681 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     7741 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     7801 acaatttcac acaggaaaca gctatgacca tgaggcgcgc cg
//
