LOCUS       sequence-107090-        6002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid of HA-tagged human Neuroligin 4 harboring a D396*
            nonsense mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6002)
  AUTHORS   Chih B, Afridi SK, Clark L, Scheiffele P
  TITLE     Disorder-associated mutations lead to functional inactivation of 
            neuroligins.
  JOURNAL   Hum Mol Genet. 2004 Jul 15;13(14):1471-7. Epub 2004 May 18.
  PUBMED    15150161
REFERENCE   2  (bases 1 to 6002)
  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: "Hum Mol 
            Genet."; date: "2004-07-15"; volume: "13(14)"; pages: "1471-7. Epub 
            2004 May 18"
COMMENT     Sequence Label: pCAG-HA-NLGN4 D396*
FEATURES             Location/Qualifiers
     source          1..6002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        368..645
                     /label=chicken beta-actin promoter
     intron          647..1664
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1588..1610
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1672..1691
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1812..1838
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3006..3025)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3071..3126
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3125..3144)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3487..3503)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3487..3503)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3500..3522)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3511..3527
                     /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)."
     promoter        complement(join(3535..3541,3542..3559,3560..3565))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3580..3601
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3660..3855
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3706..3841
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3861..3995
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3898..3917)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3952..3971
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4063..4080)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4234..4822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4314..4333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4993..5784,5785..5853))
                     /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     5616..5635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5854..5958)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(5989..6002,1..366)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca
      421 cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg
      481 gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg
      541 agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg
      601 cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcggggag tcgctgcgac
      661 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
      721 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt
      781 agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc
      841 tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg
      901 tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg
      961 cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc
     1021 ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt
     1081 gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag
     1141 ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg
     1201 ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg
     1261 ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg
     1321 cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt
     1381 tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc
     1441 gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt
     1501 cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct
     1561 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
     1621 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
     1681 tggttattgt gctgtctcat cattttggca aagaattcgc ggccgctagc accatggctg
     1741 ctggctgcct gctggcctgg acgctgacac tttggcaggc gtggctgatc ctgatcgggc
     1801 cctcgtcgga gtacccatac gacgtcccag actacgctgg tacctatcca gttgtcaaca
     1861 caaattatgg caaaatccgg ggcctaagaa caccgttacc caatgagatc ttgggtccag
     1921 tggagcagta cttaggggtc ccctatgcct caccccccac tggagagagg cggtttcagc
     1981 ccccagaacc cccgtcctcc tggactggca tccgaaatac tactcagttt gctgctgtgt
     2041 gcccccagca cctggatgag agatccttac tgcatgacat gctgcccatc tggtttaccg
     2101 ccaatttgga tactttgatg acctatgttc aagatcaaaa tgaagactgc ctttacttaa
     2161 acatctacgt gcccacggaa gatgatattc atgatcagaa cagtaagaag cccgtcatgg
     2221 tctatatcca tgggggatct tacatggagg gcaccggcaa catgattgac ggcagcattt
     2281 tggcaagcta cggaaacgtc attgtgatca ccattaacta ccgtctggga atactagggt
     2341 ttttaagtac cggtgaccag gcagcaaaag gcaactatgg gctcctggat cagattcaag
     2401 cactgcggtg gattgaggag aatgtgggag cctttggcgg ggaccccaag agagtgacca
     2461 tctttggctc gggggctggg gcctcctgtg tcagcctgtt gaccctgtcc cactactcag
     2521 aaggtctctt ccagaaggcc atcattcaga gcggcaccgc cctgtccagc tgggcagtga
     2581 actaccagcc ggccaagtac actcggatat tggcagacaa ggtcggctgc aacatgctgg
     2641 acaccacgga catggtagaa tgcctgcgga acaagaacta caaggagctc atccagcaga
     2701 ccatcacccc ggccacctac cacatagcct tcgggccggt gatcgacggc gacgtcatcc
     2761 cagacgaccc ccagatcctg atggagcaag gcgagttcct caactacgac atcatgctgg
     2821 gcgtcaacca aggggaaggc ctgaagttcg tggacggcat cgtggataac gaggacggtg
     2881 tgacgcccaa cgacttttga ctcgaggtac ccccggggag ctcaattcac tcctcaggtg
     2941 caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac
     3001 tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     3061 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     3121 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     3181 gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa
     3241 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     3301 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     3361 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     3421 gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta
     3481 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     3541 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     3601 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg
     3661 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3721 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3781 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3841 ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca
     3901 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3961 ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg
     4021 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4081 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4141 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4201 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4261 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4321 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4381 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4441 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4501 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4561 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4621 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4681 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4741 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4801 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4861 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4921 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4981 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5041 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5101 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5161 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5221 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5281 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5341 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5401 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5461 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5521 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5581 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5641 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5701 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5761 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5821 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5881 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5941 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcga cattgattat
     6001 tg
//
