LOCUS       sequence-316163-        4662 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone containing postsynaptic HyCal component (HyNaC9)
            for HySyn system fused to eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4662)
  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 4662)
  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: pJH202_pE12_hyCal9-eGFP
FEATURES             Location/Qualifiers
     source          1..4662
                     /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"
     CDS             join(1536..1538,1539..2246)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELY"
     primer_bind     complement(1578..1599)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1839..1858)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2186..2207
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2251..2350)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2367..2386)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2368..2386)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2391..2407)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2391..2407)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2509..2528)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2520..3329
                     /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(2587..2606)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3476..4064
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3965..3984
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4218..4235
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4394..4421)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4513..4599)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4648..4662,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4662,1..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 gcggaatgag taaaggagaa gaacttttca
     1561 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1621 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     1681 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     1741 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1801 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1861 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1921 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1981 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     2041 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     2101 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     2161 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     2221 tcactctcgg catggacgag ctgtactagc acccagcttt cttgtacaaa gttggcatta
     2281 taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa
     2341 tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg
     2401 tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
     2461 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     2521 tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt
     2581 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct
     2641 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     2701 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     2761 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     2821 gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     2881 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     2941 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     3001 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     3061 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     3121 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     3181 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     3241 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     3301 ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg taacactggc
     3361 agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga
     3421 gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3481 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3541 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3601 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3661 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3721 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3781 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3841 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3901 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3961 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4021 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4081 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4141 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4201 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     4261 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     4321 actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc caggaagagt
     4381 ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt gatgcctggc
     4441 agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc
     4501 cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
     4561 aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag ttccctactc
     4621 tcgcgttaac gctagcatgg atgttttccc agtcacgacg tt
//
