LOCUS       sequence-316160-        4815 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone containing postsynaptic HyCal component (HyNaC7)
            for HySyn system fused to eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4815)
  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 4815)
  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: pJH200_pE12_hyCal7-eGFP
FEATURES             Location/Qualifiers
     source          1..4815
                     /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(1689..1691,1692..2399)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELY"
     primer_bind     complement(1731..1752)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1992..2011)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2339..2360
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2404..2503)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2520..2539)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2521..2539)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2544..2560)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2544..2560)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2662..2681)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2673..3482
                     /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(2740..2759)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3629..4217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4118..4137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4371..4388
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4547..4574)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4666..4752)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4801..4815,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4815,1..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 gagtaaagga gaagaacttt tcaccggggt ggtgcccatc ctggtcgagc
     1741 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     1801 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     1861 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     1921 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1981 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     2041 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     2101 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     2161 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     2221 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     2281 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     2341 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtact
     2401 agcacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt
     2461 gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc
     2521 cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc
     2581 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt
     2641 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga
     2701 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
     2761 atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt
     2821 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
     2881 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
     2941 atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag
     3001 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
     3061 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg
     3121 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
     3181 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt
     3241 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
     3301 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
     3361 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     3421 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
     3481 aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg
     3541 gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     3601 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3661 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3721 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3781 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3841 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3901 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3961 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4021 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4081 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4141 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4201 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4261 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4321 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4381 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     4441 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     4501 caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt
     4561 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
     4621 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
     4681 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
     4741 tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt
     4801 cccagtcacg acgtt
//
