LOCUS       sequence-328557-        4869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone containing postsynaptic HyCal component (HyNaC2)
            for HySyn system fused to mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4869)
  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 4869)
  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: pJH199_pE12_hyCal2-mCherry
FEATURES             Location/Qualifiers
     source          1..4869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     94..111
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(270..297)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(389..475)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     524..546
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     538..555
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     539..555
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    571..670
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2996..3095)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3112..3131)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3113..3131)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3136..3152)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3136..3152)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3254..3273)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3265..4074
                     /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(3332..3351)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4221..4809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4710..4729
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
      241 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
      301 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
      361 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
      421 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
      481 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt
      541 aaaacgacgg ccagtcttaa gctcgggccc caaataatga ttttattttg actgatagtg
      601 acctgttcgt tgcaacaaat tgatgagcaa tgctttttta taatgccaac tttgtacaaa
      661 aaagcaggct tagctagcgc caccatgaag gctagattcg ccagaagaat ggagaaaatc
      721 gtggaggaaa gaatgaccgt gaaacagatt tttaatagat acgtgcagac ctctaccctg
      781 cacggatttc ggttcatctt catggatacc ttcattgtga gaagagtgct gtggacaatc
      841 ctgaccctga caatggctac aattttcttt aaggaactga gaaattccat caatctgttc
      901 tatgagtatc cattcacaac aaccagcacc atccagtacg aaccatctct gacattccca
      961 gctattagcg tgtgtaatct gaatcacttt ctgctgagca aaatcaagaa gagcaaactg
     1021 aagccactgt atgatcaggg aagactgcca tttgacaaca atctggagaa tccaggattc
     1081 gatattcagg gagaggagct gtacagcatt ctgaagacaa gctctcagag cattgatgaa
     1141 atttttctgt cttgcgaatg gaaaagcaga gacacagcca agaatggagt gccaaatcca
     1201 tgcaaaccaa acaacttcac agtgtacagc ggactgtacg gacagagctg ctacacattc
     1261 aatccaggag tgagcggata tccactgctg tctctgagcg aaaccggagt gaacatggga
     1321 ttcaagctgg agctggatct gaagtctaag gactctctgc agggaattca ggagattgga
     1381 gctatcgtga tcgtgcatca ccagcaggag accccagtgc tgcaggccgg attcgtggtg
     1441 tctccaggat tccagacatt cgtggagatt aaggtgagac agaccgaaaa tctgccacca
     1501 ccatatgcta caaaatgcgg atctaaacca ctgaagaact atcagatcta cagacagtct
     1561 agctgctttc tggaacagct gggagatgcc attgaaacaa agtgcaaatg caagagctct
     1621 tttatggctg gaagaaatat cccatactgc tctctgagac agaccgtgac atgtctgatg
     1681 ccaaccatct acgactttga cagaaagaca aataacaatt gcccagtgga ctgtgaaaca
     1741 attcagtacc tgtctagcct gtcttacgct agattcatta gcaatgtgac atacctgagc
     1801 aagaacgctg aacagagcag ctacattaga aaactgaaaa atagcatgtc cccaaaaaag
     1861 ctgcagaagt atatcgaaga aaatattgtg gctgtgcagt tcttttacca ggagatgaag
     1921 aaggaaaagg tgaaacagga gccaagctac gatttttata aactgattgg agacgtggga
     1981 ggacagctgg gactgctgct gggagcctct gtgctgacac tggtggagtt tgtggatctg
     2041 ttcattttta ccctgtacca ccagctgctg agactgagca aaaagaagtc aggtggctca
     2101 ggtggtggcg gttcaggcgg aatgggatcc atggtctcaa agggtgaaga agataacatg
     2161 gcaattatta aagagtttat gcgtttcaag gtgcatatgg agggatctgt caatgggcat
     2221 gagtttgaaa ttgaaggtga aggagaaggc cgaccatatg agggaacaca aaccgcaaaa
     2281 ctaaaggtaa gtttaaacat atatatacta actaaccctg attatttaaa ttttcaggta
     2341 actaaaggcg gaccattacc attcgcctgg gacatcctct ctccacagtt catgtatgga
     2401 agtaaagctt atgttaaaca tccggcagat ataccagatt atttgaaact ttcattcccg
     2461 gagggtttta agtgggaacg cgtaatgaat tttgaagacg gaggagttgt tacagtgacg
     2521 caagactcaa ggtaagttta aacagttcgg tactaactaa ccatacatat ttaaattttc
     2581 agcctccaag atggagaatt tatttataaa gtcaaacttc gaggaacgaa tttcccctcg
     2641 gatggacctg ttatgcagaa gaagactatg ggatgggaag cttcaagtga aagaatgtac
     2701 cctgaagacg gtgctcttaa gggagagatt aaacaacgtc ttaaattgaa agatggagga
     2761 cattacgatg ctgaggtaag tttaaacatg attttactaa ctaactaatc tgatttaaat
     2821 tttcaggtga agacaactta caaagccaaa aaaccagttc agctgccagg agcgtacaat
     2881 gttaatatta aactggatat cacctcccac aacgaggatt acactatcgt tgagcaatat
     2941 gaaagagctg aagggcggca ctcgacaggt ggcatggatg aattgtataa gtagtaccca
     3001 gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa
     3061 caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt
     3121 gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc
     3181 tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac
     3241 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga
     3301 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
     3361 caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg
     3421 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
     3481 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     3541 tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct
     3601 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     3661 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca
     3721 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
     3781 gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc
     3841 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     3901 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     3961 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     4021 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa
     4081 ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc
     4141 tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta
     4201 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4261 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4321 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4381 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4441 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4501 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4561 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4621 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4681 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4741 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4801 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4861 gctcacatg
//
