LOCUS       sequence-329487-        5365 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mouse NL2 N-terminally tagged with GST.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5365)
  AUTHORS   Halff EF, Szulc BR, Lesept F, Kittler JT
  TITLE     SNX27-Mediated Recycling of Neuroligin-2 Regulates Inhibitory 
            Signaling.
  JOURNAL   Cell Rep. 2019 Nov 26;29(9):2599-2607.e6. doi: 
            10.1016/j.celrep.2019.10.096.
  PUBMED    31775031
REFERENCE   2  (bases 1 to 5365)
  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.celrep.2019.10"; journalName: "Cell Rep"; date: 
            "2019-11-26- 26"; volume: "29"; issue: "9"; pages: "2599-2607.e6"
COMMENT     Sequence Label: pGEX4T3-NL2CT
FEATURES             Location/Qualifiers
     source          1..5365
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(72..77,78..93,94..100)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    108..124
                     /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)."
     CDS             147..800
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     758..780
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             807..824
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1304..1326)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1471..1489)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1557..1661
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1662..1730,1731..2522)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1880..1899)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2693..3281
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3182..3201
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3435..3452
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3525..3602
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3603..4685
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(3622..3641)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4734..4739,4740..4757,4758..4764)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4772..4788
                     /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     4796..4812
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4796..4812
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4808..5335
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4828..4845)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4828..4844)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4837..4859)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct
       61 gaaatgagct gttgacaatt aatcatcggc tcgtataatg tgtggaattg tgagcggata
      121 acaatttcac acaggaaaca gtattcatgt cccctatact aggttattgg aaaattaagg
      181 gccttgtgca acccactcga cttcttttgg aatatcttga agaaaaatat gaagagcatt
      241 tgtatgagcg cgatgaaggt gataaatggc gaaacaaaaa gtttgaattg ggtttggagt
      301 ttcccaatct tccttattat attgatggtg atgttaaatt aacacagtct atggccatca
      361 tacgttatat agctgacaag cacaacatgt tgggtggttg tccaaaagag cgtgcagaga
      421 tttcaatgct tgaaggagcg gttttggata ttagatacgg tgtttcgaga attgcatata
      481 gtaaagactt tgaaactctc aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa
      541 tgttcgaaga tcgtttatgt cataaaacat atttaaatgg tgatcatgta acccatcctg
      601 acttcatgtt gtatgacgct cttgatgttg ttttatacat ggacccaatg tgcctggatg
      661 cgttcccaaa attagtttgt tttaaaaaac gtattgaagc tatcccacaa attgataagt
      721 acttgaaatc cagcaagtat atagcatggc ctttgcaggg ctggcaagcc acgtttggtg
      781 gtggcgacca tcctccaaaa tcggatctgg ttccgcgtgg atcctacaag cgggaccggc
      841 gccaggagct gcggtgccgg aggcttagcc caccaggagg ctcaggctca ggtgtgcctg
      901 gtgggggccc cctgcttccc actgctggcc gtgagctacc cccggaggag gagctagtat
      961 cgctgcagct gaagcggggt ggtggtgttg gggcggaccc tgctgaggcc ctgcgccctg
     1021 cctgtccacc cgactatacc ctggccttgc gccgggcacc ggacgatgtg cctctcttgg
     1081 cccccggggc cctaaccctg ctgcctagtg gcctggggcc cccgcccccg cccccacccc
     1141 cttctctcca tccctttggg cccttcccac ccccaccccc tactgctacc agccacaaca
     1201 acacgctacc ccatccccac tccaccactc gggtatagct cgagcggccg catcgtgact
     1261 gactgacgat ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     1321 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     1381 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
     1441 gcggagtgta taattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     1501 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     1561 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     1621 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     1681 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     1741 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     1801 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     1861 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     1921 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     1981 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2041 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2101 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2161 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     2221 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2281 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2341 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     2401 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     2461 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     2521 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     2581 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     2641 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2701 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2761 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2821 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2881 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2941 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3001 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3061 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3121 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3181 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3241 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3301 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     3361 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     3421 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     3481 ttctccttac gcatctgtgc ggtatttcac accgcataaa ttccgacacc atcgaatggc
     3541 gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga
     3601 atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg
     3661 tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag
     3721 cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac
     3781 agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg
     3841 tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag
     3901 aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca
     3961 gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct
     4021 gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta
     4081 ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc
     4141 agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg
     4201 gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact
     4261 ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca
     4321 ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt
     4381 ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct
     4441 catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca
     4501 gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc
     4561 ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc
     4621 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     4681 agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcacccca ggctttacac
     4741 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     4801 aacagctatg accatgatta cggattcact ggccgtcgtt ttacaacgtc gtgactggga
     4861 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     4921 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     4981 atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc tggagtgcga
     5041 tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc
     5101 gcccatctac accaacgtga cctatcccat tacggtcaat ccgccgtttg ttcccacgga
     5161 gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc tacaggaagg
     5221 ccagacgcga attatttttg atggcgttgg aattagctta tcgactgcac ggtgcaccaa
     5281 tgcttctggc gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac
     5341 tgcataattc gtgtcgctca aggcg
//
