LOCUS       sequence-248787-        4675 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express HA tagged Nxph4-4Q.
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 Apr 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-Nxph4-4Q-HA
FEATURES             Location/Qualifiers
     source          1..4675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..330
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        331..534
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     485..505
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     531..555
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          672..768
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             1773..1799
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1814..1948
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1838..1857)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1892..1911
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2059..2075)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2059..2075)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2072..2094)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2083..2099
                     /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(2107..2113,2114..2131,2132..2137))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2152..2173
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2290..2307)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2461..3049)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2541..2560)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3220..4011,4012..4080))
                     /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     3843..3862
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4081..4185)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4253..4271
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4309..4331)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4431..4450
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4644..4666
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4658..4675
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4659..4675
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gagttcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc
       61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      541 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
      601 cgggaccgat ccagcctccg gactctagag gatccggtac tagaggaact gaaaaaccag
      661 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg
      721 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa
      781 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg ggcccaggcc cgaattcggt
      841 cgaccatgcg gctgctcccg gaatggctcc tcttgctctt tggtccttgg ctccttagga
      901 aggctgtctg tggccagatc gtggagtccg ggaggcccca gtacctagac ttgcgtcccg
      961 ccatggccgg agggggcgcc cgcggccagc agctcccagc gcccgcatct tccgagggcc
     1021 tgggcatggc gcgctcctgg agctgggctt ggccggctaa ccacaccggg gcgctggcca
     1081 agccagggac agccggtggc ccgtccgtgc cgcgcaccaa gaggaagcca tccattaaag
     1141 cggcccgagc caagaagatt ttcggctggg gggactttta ctttcgtgtg cataccctca
     1201 agttctcgct gttggtgacc ggcaagatcg tggaccatgt gaacggtacc ttcagtgtgt
     1261 acttccgtca caattcgtcc agcctgggta acctaagcgt cagtatcgtg ccgccctcca
     1321 agcgtgtgga gttcgggggc gtctggttgc ctgggcctgc ctcccaccct ctacagtcca
     1381 cgctggcact tgaaggggtg cttcctggat tggggccccc tctggggatg gcggggcagg
     1441 ggctggggag caaccttggg ggcgcacttg ccggcccact cggaagcgcg ctaggagttc
     1501 ctggggccaa agagtcgcgc gcttttaatg cccacgtgga atatgagaag acaaaccgcg
     1561 ctcgcaagca ccgcccggcc ctgtacgacc cgtcgcaggt ggccttcacc gagcacacgc
     1621 agagccaggc cgcctggctc gccgccaagc ccttcaaagt catcgctatc ttcgtctcct
     1681 tcctcagttt tgactacaaa ctggtgcaga aggtggcccc agactataac ttccagagtg
     1741 agcaccccta ctttggatct cgaggtaccg gctacccata cgatgttcca gattacgctt
     1801 aagcggccgc ggggatccag acatgataag atacattgat gagtttggac aaaccacaac
     1861 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     1921 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     1981 ggttcagggg gaggtgtggg aggttttttc ggatcctcta gagtcgatct gcaggcatgc
     2041 tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2101 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     2161 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2221 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2281 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2341 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2401 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2461 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2521 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2581 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2641 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2701 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2761 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2821 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2881 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     2941 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3001 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3061 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3121 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3181 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3241 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3301 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3361 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3421 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3481 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3541 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3601 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3661 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3721 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3781 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3841 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3901 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3961 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4021 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4081 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4141 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4201 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     4261 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     4321 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     4381 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     4441 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     4501 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     4561 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     4621 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagt
//
