LOCUS       sequence-263525-        6538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter expressing membrane targeted EGFP for visualization of 
            neuronal morphology upon Cre-based recombination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6538)
  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 6538)
  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-loxSTOPlox-LynEGFP
FEATURES             Location/Qualifiers
     source          1..6538
                     /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    1741..1774
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1941..2075
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2195..2329
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2449..2583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    2612..2645
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2671..2680
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2722..2724,2725..2727,2728..3441)
                     /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(2722..2724,2725..2727,2728..3438)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2767..2788)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3028..3047)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3375..3396
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3457..3474)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3463..3687
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3759..3776)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3759..3775)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3768..3790)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3916..4371)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4053..4074)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4265..4284
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4398..4502
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4503..4571,4572..5363)
                     /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(4721..4740)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5534..6122
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6023..6042
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6276..6293
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6410..6431
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6446..6451,6452..6469,6470..6476)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6484..6500
                     /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     6489..6511
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6508..6524
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6508..6524
                     /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 gsgaggcgga 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 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 ccgccaccaa tcatttaaat
     1741 ataacttcgt ataatgtatg ctatacgaag ttattcgcga tgaataaatg aaagcttgca
     1801 gatctgcgac tctagaggat ctgcgactct agaggatcat aatcagccat accacatttg
     1861 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     1921 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     1981 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     2041 ccaaactcat caatgtatct tatcatgtct ggatctgcga ctctagagga tcataatcag
     2101 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2161 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2221 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2281 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct gcgactctag
     2341 aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     2401 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     2461 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2521 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     2581 atccccatca agctgatccg gaacccttaa tataacttcg tataatgtat gctatacgaa
     2641 gttattaggt ccctcgacct gcagctgcag gccaccatgg gatgtattaa atcaaaaagg
     2701 aaagacgggg cccggtacca tatggtgagc aagggcgagg agctgttcac cggggtggtg
     2761 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     2821 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2881 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     2941 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     3001 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     3061 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     3121 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     3181 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     3241 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     3301 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     3361 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3421 atggacgagc tgtacaagta agagctcgct gatcagcctc gactgtgcct tctagttgcc
     3481 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     3541 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3601 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     3661 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga
     3721 gatccactag ttctagcctc gaggctagag cggccgccac tggccgtcgt tttacaacgt
     3781 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     3841 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     3901 ctgaatggcg aatggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3961 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     4021 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4081 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4141 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4201 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4261 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     4321 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     4381 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4441 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4501 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4561 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4621 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4681 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4741 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4801 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4861 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4921 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4981 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5041 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5101 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5161 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5221 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5281 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5341 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5401 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5461 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5521 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5581 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5641 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5701 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5761 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5821 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5881 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5941 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6001 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6061 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6121 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6181 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6241 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     6301 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     6361 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     6421 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     6481 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgag gcgcgccg
//
