LOCUS       sequence-316164-        4821 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone containing postsynaptic HyCal component (HyNaC9)
            for HySyn system fused to mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4821)
  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 4821)
  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: pJH203_pE12_hyCal9-mCherry
FEATURES             Location/Qualifiers
     source          1..4821
                     /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(2410..2509)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2526..2545)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2527..2545)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2550..2566)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2550..2566)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2668..2687)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2679..3488
                     /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(2746..2765)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3635..4223
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4124..4143
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4377..4394
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4553..4580)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4672..4758)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4807..4821,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4821,1..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 gcgaggaaga tgagaagaaa attaagagcc
      181 cacatgaaca gagaaatgat agagtgaaag aacacgtggc tcatctgatt aagaatgtga
      241 gcttccatgg actgagctat gtggctgata aaagaaacaa ctacttcaga agagctattt
      301 ggttcctgat caccgtggga gccttcatct atgctgtgga aaaggtgtat gagtctaccg
      361 tgaattactt tagctatcca ttcaagacag ctagaatgaa aatctatgtg aatgaactga
      421 acttcccagc tgtgagcttc tgcaatctga acgattttct gttttctaag ctgaatggaa
      481 ccaaactgga tgaaagcatc ctgtacccag atgatccaga gaaaaacaat gtgtctgaaa
      541 ttgagatttc caatatcaca tctgacgcta caatcagact ggatcagatg ctggtggatt
      601 gcgaattcga gggaaaaaag tgtacacacg agaatttcac cgatttctgg accatgcagg
      661 gagagctgtg cttcacattc aatagcggaa aaaattctca ctctctgctg aaggtgtctg
      721 gagtgggagt gctgagatct ctgaagctga ccattaacgt gcagcattat gagtattaca
      781 gagatgagat ggccggagga attcacctga tgattcatgg acaggatgaa gagccagtga
      841 aaatgcaggg acagatcgtg tctccaggat attctttcta tgtgaaggtg gaaaagaaga
      901 ccatcatgaa cctggagaaa ccatataaga cagagtgcgg aaccgtgaaa ctgaaatatt
      961 ttgatagata ctccatgcac acatgctggc tggagcagct gaccgattat gtgaataaga
     1021 tgtgccactg caaggacttc ttcatgccag gaaatattcc atattgttct ctgccagaac
     1081 tgcagaactg tacctggatt gagtgggcta aattcaataa ggataaaatg tacaagtgcc
     1141 cactgccatg taagattgac ctgtacggag tgtctctgtc tagagccctg ttcccaacca
     1201 cacagtatag ctctatcctg gccgagcagt tcagaaaaca gccacatgtg ctgtctatcg
     1261 tgcacaacat tacagatgag ctgctgttca tgagagataa cctgctgaga ttcattatct
     1321 attacgatga tctgagctat gaggtgctgg aacagaaacc atcttatgag acactggtgt
     1381 ggctgggaga cattggagga cagatcggac tgtttatcgg agccggagtg atgagctact
     1441 tcgagttcct ggattgcctg gccatcgtga tctacaccag atttttccag aagttcacaa
     1501 gcagcggtgg ctcaggtggt ggcggttcag gcggaatggg atccatggtc tcaaagggtg
     1561 aagaagataa catggcaatt attaaagagt ttatgcgttt caaggtgcat atggagggat
     1621 ctgtcaatgg gcatgagttt gaaattgaag gtgaaggaga aggccgacca tatgagggaa
     1681 cacaaaccgc aaaactaaag gtaagtttaa acatatatat actaactaac cctgattatt
     1741 taaattttca ggtaactaaa ggcggaccat taccattcgc ctgggacatc ctctctccac
     1801 agttcatgta tggaagtaaa gcttatgtta aacatccggc agatatacca gattatttga
     1861 aactttcatt cccggagggt tttaagtggg aacgcgtaat gaattttgaa gacggaggag
     1921 ttgttacagt gacgcaagac tcaaggtaag tttaaacagt tcggtactaa ctaaccatac
     1981 atatttaaat tttcagcctc caagatggag aatttattta taaagtcaaa cttcgaggaa
     2041 cgaatttccc ctcggatgga cctgttatgc agaagaagac tatgggatgg gaagcttcaa
     2101 gtgaaagaat gtaccctgaa gacggtgctc ttaagggaga gattaaacaa cgtcttaaat
     2161 tgaaagatgg aggacattac gatgctgagg taagtttaaa catgatttta ctaactaact
     2221 aatctgattt aaattttcag gtgaagacaa cttacaaagc caaaaaacca gttcagctgc
     2281 caggagcgta caatgttaat attaaactgg atatcacctc ccacaacgag gattacacta
     2341 tcgttgagca atatgaaaga gctgaagggc ggcactcgac aggtggcatg gatgaattgt
     2401 ataagtagta cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa
     2461 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     2521 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
     2581 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
     2641 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     2701 cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     2761 gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc
     2821 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     2881 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     2941 ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat
     3001 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     3061 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     3121 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     3181 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
     3241 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     3301 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     3361 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     3421 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     3481 ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac
     3541 gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag
     3601 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3661 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3721 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3781 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3841 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3901 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3961 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4021 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4081 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4141 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4201 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4261 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     4321 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     4381 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     4441 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     4501 gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc
     4561 atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc
     4621 ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct
     4681 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg
     4741 agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga
     4801 tgttttccca gtcacgacgt t
//
