LOCUS       sequence-316155-        4079 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone containing postsynaptic HyCal component (HyNaC7)
            for HySyn system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4079)
  AUTHORS   Hawk JD, Wisdom EM, Sengupta T, Kashlan ZD, Colon-Ramos DA
  TITLE     A genetically encoded tool for reconstituting synthetic modulatory 
            neurotransmission and reconnect neural circuits in vivo.
  JOURNAL   Nat Commun. 2021 Aug 9;12(1):4795. doi: 10.1038/s41467-021-24690-9.
  PUBMED    34373460
REFERENCE   2  (bases 1 to 4079)
  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: "Nat 
            Commun."; date: "2021-08-9"; volume: "12(1):4795. doi"; pages: " 
            10.1038/s41467-021-24690-9"
COMMENT     Sequence Label: pJH54_pE12_hyCal7
FEATURES             Location/Qualifiers
     source          1..4079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1668..1767)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1784..1803)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1785..1803)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1808..1824)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1808..1824)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1926..1945)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1937..2746
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2004..2023)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2893..3481
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3382..3401
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3635..3652
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3811..3838)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3930..4016)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4065..4079,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4079,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagcagg cttagctagc gccaccatga gaatcagact gaaggagaga atgagaaaaa
      181 ttgtgcagga aagactgacc gtgaaacaga tttttaaaag atatatcgag agctctaccc
      241 tgcacggatt ttgttatgtg tgtatggaca cattcctggg aagaagactg atttgggccg
      301 tgctgatgat cctgggagcc atctatttca tttttaaact gagatacggc atcaaggaat
      361 actttgacta tccattcagc accctgtcta cagtggaata tgtggacgat ctgctgtttc
      421 cagccgtgtc tgtgtgcgct accaatagct acattgcctc tcaggtgtat accaatcagc
      481 tgaatacaat gtataaggag ggaagactgc cactggataa caaccagtct atcccagaat
      541 ataacattcc aggagacgag ctggtgaaga ccctgaagaa cagcagcctg acaattgagt
      601 ctctgctgaa gtattgtgac tggatcatgc aggacacatc tcacccactg gtgaccccaa
      661 ataattgtgg agctctgaat tttacaagct attttaatta caagggagag cagtgccata
      721 ccctgaacag cggagctaaa ggacatgaac tgctgaaggt gagcgatgtg ggaatctctc
      781 atggatacga actggtgttt gacctgcaga caaacgaggt tattaagaac tatcagctga
      841 gcggaatgag aatcgtgatc catgatcagg tgttcccacc acagctggtg gatggattct
      901 ttatcagccc aggattcaag acctacatca agctgggaat tacccagtct cagtctctgc
      961 caccaccata ttctacagaa tgcggacaga aaaaactgaa gtattatgct atctatagcc
     1021 agagactgtg cctgctggag accctgacag acttcacagg agacctgtgc ggatgtagag
     1081 atgtgtttat gccagagaac ggactgccat tctgtagcct gcaggaactg tatagctgca
     1141 tgtacccagc taaagagtct tttagcgaat tcaccatgag aaaagagtgt ccaagcgatt
     1201 gcgaagaaag aacctttagc tacgagctgt ctgaggctag atatctgcac aatccaccaa
     1261 ttggactgtc tctgagcaga ctggataaac tgcagctgga caagtctctg ccaagcgagg
     1321 ctcacctgtc taaactggct aaaagcctga caccaaaaga actggatgcc tacatcgaaa
     1381 gcaacatcat cagcgtgatt ctgtttttcg gagataccag aatcgattac cacgaacagg
     1441 aagccacaaa cgactttttc cagttcctgg gaaatatggg aggagaattt ggactgatgc
     1501 tgggagcttc cctgctgacc ttcgtggaat tcattgacct gtttattgtg ctgctgtacc
     1561 accagatgct gagactgtat aacctgagaa agattccaga catttttgcc agcaaaagaa
     1621 acagaaataa ggaaaaaacc aagaagtgtg agaacgtgtg aggatccacc cagctttctt
     1681 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     1741 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     1801 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     1861 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     1921 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     1981 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2041 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     2101 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     2161 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     2221 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     2281 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     2341 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     2401 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     2461 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     2521 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     2581 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     2641 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     2701 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     2761 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     2821 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     2881 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2941 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3001 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3061 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3121 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3181 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3241 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3301 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3361 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3421 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3481 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3541 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     3601 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     3661 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     3721 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac
     3781 cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc
     3841 ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc
     3901 ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac
     3961 aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc
     4021 ctggcagttc cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgtt
//
