LOCUS       sequence-248758-        4675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HA tagged Nxph4-6A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4675)
  AUTHORS   Meng X, McGraw CM, Wang W, Jing J, Yeh SY, Wang L, Lopez J, Brown 
            AM, Lin T, Chen W, Xue M, Sillitoe RV, Jiang X, Zoghbi HY
  TITLE     Neurexophilin4 is a selectively expressed alpha-neurexin ligand that
            modulates specific cerebellar synapses and motor functions.
  JOURNAL   Elife. 2019 Sep 16;8. pii: 46773. doi: 10.7554/eLife.46773.
  PUBMED    31524598
REFERENCE   2  (bases 1 to 4675)
  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: "Elife."; 
            date: "2019-09-16"; volume: "8. pii: 46773. doi"; pages: " 
            10.7554/eLife.46773"
COMMENT     Sequence Label: pCMV-Nxph4-6A-HA
FEATURES             Location/Qualifiers
     source          1..4675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        44..347
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        348..551
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     502..522
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     548..572
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          689..785
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             1790..1816
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1831..1965
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1855..1874)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1909..1928
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2076..2092)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2076..2092)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2089..2111)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2100..2116
                     /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(2124..2130,2131..2148,2149..2154))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2169..2190
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2307..2324)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2478..3066)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2558..2577)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3237..4028,4029..4097))
                     /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     3860..3879
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4098..4202)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4270..4288
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4326..4348)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4448..4467
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4661..4675,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4675,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgag ttcgagcttg catgcctgca ggtcgttaca taacttacgg
       61 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      121 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      181 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      241 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      301 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      361 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      421 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      481 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      541 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
      601 acctccatag aagacaccgg gaccgatcca gcctccggac tctagaggat ccggtactag
      661 aggaactgaa aaaccagaaa gttaactggt aagtttagtc tttttgtctt ttatttcagg
      721 tcccggatcc ggtggtggtg caaatcaaag aactgctcct cagtggatgt tgcctttact
      781 tctaggcctg tacggaagtg ttacttctgc tctaaaagct gcggaattgt acccgcgggc
      841 ccaggcccga attcggtcga ccatgcggct gctcccggaa tggctcctct tgctctttgg
      901 tccttggctc cttaggaagg ctgtctgtgg ccagatcgtg gagtccggga ggccccagta
      961 cctagacttg cgtcccgcca tggccggagg gggcgcccgc ggccagcagc tcccagcgcc
     1021 cgcatcttcc gagggcctgg gcatggcgcg ctcctggagc tgggcttggc cggctaacca
     1081 caccggggcg ctggccaagc cagggacagc cggtggcccg tccgtgccgc gcaccaagag
     1141 gaagccatcc attaaagcgg cccgagccaa gaagattttc ggctgggggg acttttactt
     1201 tcgtgtgcat accctcaagt tctcgctgtt ggtgaccggc aagatcgtgg accatgtgaa
     1261 cggtaccttc agtgtgtact tccgtcacaa ttcgtccagc ctgggtaacc taagcgtcag
     1321 tatcgtgccg ccctccaagc gtgtggagtt cgggggcgtc tggttgcctg ggcctgcctc
     1381 ccaccctcta cagtccacgc tggcacttga aggggtgctt cctggattgg ggccccctct
     1441 ggggatggcg gggcaggggc tggggagcaa ccttgggggc gcacttgccg gcccactcgg
     1501 aagcgcgcta ggagttcctg gggccaaaga gtcgcgcgct tttaatgccc acgtggaata
     1561 tgagaagaca aaccgcgctc gcaagcaccg cccggccctg tacgacccgt cgcaggtggc
     1621 cttcaccgag cacacgcaga gccaggccgc ctggctcgcc gccaagccct tcaaagtcat
     1681 cgctatcttc gtctccttcc tcagttttga ctacaaactg gtgcagaagg tggccccaga
     1741 ctataacttc cagagtgagc acccctactt tggatctcga ggtaccggct acccatacga
     1801 tgttccagat tacgcttaag cggccgcggg gatccagaca tgataagata cattgatgag
     1861 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     1921 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     1981 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttcgga tcctctagag
     2041 tcgatctgca ggcatgctag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     2101 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     2161 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     2221 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2281 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2341 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2401 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2461 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2521 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2581 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2641 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2701 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2761 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2821 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2881 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2941 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3001 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3061 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3121 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3181 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3241 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3301 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3361 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3421 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3481 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3541 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3601 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3661 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3721 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3781 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3841 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3901 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3961 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4021 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4081 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4141 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4201 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     4261 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     4321 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     4381 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     4441 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     4501 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
     4561 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     4621 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgtt
//
