LOCUS       sequence-263481-        6513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter expressing membrane targeted EGFP for visualization of 
            neuronal morphology upon Dre-based recombination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6513)
  AUTHORS   Dupraz S, Hilton BJ, Husch A, Santos TE, Coles CH, Stern S, 
            Brakebusch C, Bradke F
  TITLE     RhoA Controls Axon Extension Independent of Specification in the 
            Developing Brain.
  JOURNAL   Curr Biol. 2019 Nov 18;29(22):3874-3886.e9. doi: 
            10.1016/j.cub.2019.09.040. Epub 2019 Oct 31.
  PUBMED    31679934
REFERENCE   2  (bases 1 to 6513)
  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.cub.2019.09.040"; journalName: "Curr Biol"; date: 
            "2019-11-18- 18"; volume: "29"; issue: "22"; pages: "3874-3886.e9"
COMMENT     Sequence Label: pCAG-roxSTOProx-LynEGFP
FEATURES             Location/Qualifiers
     source          1..6513
                     /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"
     primer_bind     1694..1713
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1694..1712
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1729..1760
                     /label=rox
                     /bound_moiety="Dre recombinase"
     polyA_signal    1927..2061
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2181..2315
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2435..2569
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    2598..2629
                     /label=rox
                     /bound_moiety="Dre recombinase"
     regulatory      2646..2655
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2697..2699,2700..2702,2703..3416)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2697..2699,2700..2702,2703..3413)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2742..2763)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3003..3022)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3350..3371
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3432..3449)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3438..3662
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3734..3751)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3734..3750)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3743..3765)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3891..4346)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4028..4049)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4240..4259
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4373..4477
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4478..4546,4547..5338)
                     /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(4696..4715)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5509..6097
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5998..6017
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6251..6268
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6385..6406
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6421..6426,6427..6444,6445..6451)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6459..6475
                     /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     6464..6486
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6483..6499
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6483..6499
                     /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 cgcgccaggc ggggcggggc ggggcgaggg
      541 gcggggcggg gcgaggcgga 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 taataatacg actcactata gggggtcgac ccgccaccta actttaaata
     1741 attggcatta tttaaagtta tcgcgatgaa taaatgaaag cttgcagatc tgcgactcta
     1801 gaggatctgc gactctagag gatcataatc agccatacca catttgtaga ggttttactt
     1861 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     1921 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     1981 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2041 gtatcttatc atgtctggat ctgcgactct agaggatcat aatcagccat accacatttg
     2101 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     2161 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     2221 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     2281 ccaaactcat caatgtatct tatcatgtct ggatctgcga ctctagagga tcataatcag
     2341 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2401 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2461 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2521 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc ccatcaagct
     2581 gatccggaac ccttaattaa ctttaaataa ttggcattat ttaaagttag gcagccgccc
     2641 tgcaggccac catgggatgt attaaatcaa aaaggaaaga cggggcccgg taccatatgg
     2701 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     2761 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     2821 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     2881 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     2941 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     3001 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     3061 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     3121 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     3181 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     3241 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     3301 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     3361 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagagc
     3421 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3481 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3541 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3601 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3661 ggcttctgag gcggaaagaa ccagctgggg ctcgagatcc actagttcta gcctcgaggc
     3721 tagagcggcc gccactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     3781 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     3841 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg aaattgtaag
     3901 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     3961 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     4021 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     4081 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     4141 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     4201 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     4261 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     4321 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     4381 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4441 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4501 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4561 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4621 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4681 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4741 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4801 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4861 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4921 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4981 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5041 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5101 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5161 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5221 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5281 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5341 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5401 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5461 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5521 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5581 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5641 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5701 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5761 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5821 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5881 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5941 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6001 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6061 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6121 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6181 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     6241 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     6301 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     6361 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     6421 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     6481 cacaggaaac agctatgacc atgaggcgcg ccg
//
