LOCUS       sequence-316162-        4974 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone containing postsynaptic HyCal component (HyNaC7)
            for HySyn system fused to mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4974)
  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 4974)
  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: pJH201_pE12_hyCal7-mCherry
FEATURES             Location/Qualifiers
     source          1..4974
                     /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(2563..2662)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2679..2698)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2680..2698)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2703..2719)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2703..2719)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2821..2840)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2832..3641
                     /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(2899..2918)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3788..4376
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4277..4296
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4530..4547
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4706..4733)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4825..4911)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4960..4974,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4974,1..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 agaacgtggg tggctcaggt ggtggcggtt
     1681 caggcggaat gggatccatg gtctcaaagg gtgaagaaga taacatggca attattaaag
     1741 agtttatgcg tttcaaggtg catatggagg gatctgtcaa tgggcatgag tttgaaattg
     1801 aaggtgaagg agaaggccga ccatatgagg gaacacaaac cgcaaaacta aaggtaagtt
     1861 taaacatata tatactaact aaccctgatt atttaaattt tcaggtaact aaaggcggac
     1921 cattaccatt cgcctgggac atcctctctc cacagttcat gtatggaagt aaagcttatg
     1981 ttaaacatcc ggcagatata ccagattatt tgaaactttc attcccggag ggttttaagt
     2041 gggaacgcgt aatgaatttt gaagacggag gagttgttac agtgacgcaa gactcaaggt
     2101 aagtttaaac agttcggtac taactaacca tacatattta aattttcagc ctccaagatg
     2161 gagaatttat ttataaagtc aaacttcgag gaacgaattt cccctcggat ggacctgtta
     2221 tgcagaagaa gactatggga tgggaagctt caagtgaaag aatgtaccct gaagacggtg
     2281 ctcttaaggg agagattaaa caacgtctta aattgaaaga tggaggacat tacgatgctg
     2341 aggtaagttt aaacatgatt ttactaacta actaatctga tttaaatttt caggtgaaga
     2401 caacttacaa agccaaaaaa ccagttcagc tgccaggagc gtacaatgtt aatattaaac
     2461 tggatatcac ctcccacaac gaggattaca ctatcgttga gcaatatgaa agagctgaag
     2521 ggcggcactc gacaggtggc atggatgaat tgtataagta gtacccagct ttcttgtaca
     2581 aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca
     2641 gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca
     2701 tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg
     2761 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     2821 caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca
     2881 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     2941 caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     3001 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     3061 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     3121 ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     3181 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     3241 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     3301 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     3361 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
     3421 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     3481 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     3541 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     3601 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt
     3661 tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat
     3721 cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3781 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3841 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3901 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3961 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4021 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4081 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4141 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4201 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4261 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4321 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4381 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4441 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4501 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4561 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     4621 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta
     4681 gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt
     4741 ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac
     4801 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca
     4861 acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc
     4921 agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgtt
//
