LOCUS       sequence-316157-        3926 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone containing postsynaptic HyCal component (HyNaC9)
            for HySyn system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3926)
  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 3926)
  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: pJH55_pE12_hyCal9
FEATURES             Location/Qualifiers
     source          1..3926
                     /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(1515..1614)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1631..1650)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1632..1650)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1655..1671)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1655..1671)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1773..1792)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1784..2593
                     /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(1851..1870)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2740..3328
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3229..3248
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3482..3499
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3658..3685)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3777..3863)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3912..3926,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3926,1..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 gcagctgagg atccacccag ctttcttgta caaagttggc attataagaa agcattgctt
     1561 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     1621 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     1681 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     1741 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     1801 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     1861 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     1921 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     1981 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     2041 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     2101 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     2161 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     2221 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     2281 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     2341 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     2401 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     2461 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     2521 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     2581 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     2641 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     2701 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2761 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2821 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2881 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2941 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3001 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3061 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3121 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3181 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3241 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3301 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3361 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3421 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3481 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     3541 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     3601 tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa
     3661 aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt
     3721 cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt
     3781 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc
     3841 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc
     3901 atggatgttt tcccagtcac gacgtt
//
