LOCUS       sequence-372390-        6227 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  membrane target protein labelling .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6227)
  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 6227)
  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-loxpSTOPloxp-lyn-NeonGreen
FEATURES             Location/Qualifiers
     source          1..6227
                     /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    1693..1726
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1893..2027
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2147..2281
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    2310..2343
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2369..2378
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2420..3130
                     /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      2675..2684
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3146..3163)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3152..3376
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3448..3465)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3448..3464)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3457..3479)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3605..4060)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3742..3763)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3954..3973
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4087..4191
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4192..4260,4261..5052)
                     /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(4410..4429)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5223..5811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5712..5731
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5965..5982
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6099..6120
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6135..6140,6141..6158,6159..6165)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6173..6189
                     /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     6178..6200
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6197..6213
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6197..6213
                     /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 cgcgccaggs ggggsggggs ggggcgrggg
      541 gsggggsggg 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 gatccttaaa atataacttc gtataatgta tgctatacga agttattcgc gatgaataaa
     1741 tgaaagcttg cagatctgcg actctagagg atctgcgact ctagaggatc ataatcagcc
     1801 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     1861 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     1921 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     1981 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatctgc gactctagag
     2041 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     2101 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     2161 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     2221 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     2281 ccccatcaag ctgatccgga acccttaata taacttcgta taatgtatgc tatacgaagt
     2341 tattaggtcc ctcgacctgc agctgcaggc caccatggga tgtattaaat caaaaaggaa
     2401 agacggggcc cggtaccata tggtgagcaa gggcgaggag gataacatgg cctctctccc
     2461 agcgacacat gagttacaca tctttggctc catcaacggt gtggactttg acatggtggg
     2521 tcagggcacc ggcaatccaa atgatggtta tgaggagtta aacctgaagt ccaccaaggg
     2581 tgacctccag ttctccccct ggattctggt ccctcatatc gggtatggct tccatcagta
     2641 cctgccctac cctgacggga tgtcgccttt ccaggccgcc atggtagatg gctccggcta
     2701 ccaagtccat cgcacaatgc agtttgaaga tggtgcctcc cttactgtta actaccgcta
     2761 cacctacgag ggaagccaca tcaaaggaga ggcccaggtg aaggggactg gtttccctgc
     2821 tgacggtcct gtgatgacca actcgctgac cgctgcggac tggtgcaggt cgaagaagac
     2881 ttaccccaac gacaaaacca tcatcagtac ctttaagtgg agttacacca ctggaaatgg
     2941 caagcgctac cggagcactg cgcggaccac ctacaccttt gccaagccaa tggcggctaa
     3001 ctatctgaag aaccagccga tgtacgtgtt ccgtaagacg gagctcaagc actccaagac
     3061 cgagctcaac ttcaaggagt ggcaaaaggc ctttaccgat gtgatgggca tggacgagct
     3121 gtacaagtaa gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3181 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3241 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3301 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3361 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctcgag atccactagt
     3421 tctagcctcg aggctagagc ggccgccact ggccgtcgtt ttacaacgtc gtgactggga
     3481 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     3541 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     3601 atggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     3661 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     3721 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     3781 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     3841 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     3901 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     3961 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     4021 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     4081 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4141 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4201 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4261 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4321 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4381 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     4441 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4501 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4561 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4621 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4681 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4741 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4801 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4861 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4921 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4981 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5041 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5101 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5161 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5221 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5281 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5341 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     5401 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5461 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5521 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5581 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5641 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5701 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5761 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5821 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5881 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     5941 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     6001 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     6061 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     6121 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     6181 ggataacaat ttcacacagg aaacagctat gaccatgagg cgcgccg
//
