LOCUS       sequence-310646-        4654 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 4654)
  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 4654)
  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..4654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(2072..2074,2075..2077,2078..2788)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     CDS             join(2072..2074,2075..2077,2078..2785)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(2117..2138)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2378..2397)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2725..2746
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2895..2914)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2896..2914)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2919..2935)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2919..2935)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3037..3056)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3048..3857
                     /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(3115..3134)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4004..4592
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4493..4512
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctta gctagcgcca ccatgagcga ggaagatgag aagaaaatta agagcccaca
      721 tgaacagaga aatgatagag tgaaagaaca cgtggctcat ctgattaaga atgtgagctt
      781 ccatggactg agctatgtgg ctgataaaag aaacaactac ttcagaagag ctatttggtt
      841 cctgatcacc gtgggagcct tcatctatgc tgtggaaaag gtgtatgagt ctaccgtgaa
      901 ttactttagc tatccattca agacagctag aatgaaaatc tatgtgaatg aactgaactt
      961 cccagctgtg agcttctgca atctgaacga ttttctgttt tctaagctga atggaaccaa
     1021 actggatgaa agcatcctgt acccagatga tccagagaaa aacaatgtgt ctgaaattga
     1081 gatttccaat atcacatctg acgctacaat cagactggat cagatgctgg tggattgcga
     1141 attcgaggga aaaaagtgta cacacgagaa tttcaccgat ttctggacca tgcagggaga
     1201 gctgtgcttc acattcaata gcggaaaaaa ttctcactct ctgctgaagg tgtctggagt
     1261 gggagtgctg agatctctga agctgaccat taacgtgcag cattatgagt attacagaga
     1321 tgagatggcc ggaggaattc acctgatgat tcatggacag gatgaagagc cagtgaaaat
     1381 gcagggacag atcgtgtctc caggatattc tttctatgtg aaggtggaaa agaagaccat
     1441 catgaacctg gagaaaccat ataagacaga gtgcggaacc gtgaaactga aatattttga
     1501 tagatactcc atgcacacat gctggctgga gcagctgacc gattatgtga ataagatgtg
     1561 ccactgcaag gacttcttca tgccaggaaa tattccatat tgttctctgc cagaactgca
     1621 gaactgtacc tggattgagt gggctaaatt caataaggat aaaatgtaca agtgcccact
     1681 gccatgtaag attgacctgt acggagtgtc tctgtctaga gccctgttcc caaccacaca
     1741 gtatagctct atcctggccg agcagttcag aaaacagcca catgtgctgt ctatcgtgca
     1801 caacattaca gatgagctgc tgttcatgag agataacctg ctgagattca ttatctatta
     1861 cgatgatctg agctatgagg tgctggaaca gaaaccatct tatgagacac tggtgtggct
     1921 gggagacatt ggaggacaga tcggactgtt tatcggagcc ggagtgatga gctacttcga
     1981 gttcctggat tgcctggcca tcgtgatcta caccagattt ttccagaagt tcacaagcag
     2041 cggtggctca ggtggtggcg gttcaggcgg aatggtgagc aagggcgagg agctgttcac
     2101 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2161 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     2221 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     2281 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     2341 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     2401 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2461 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     2521 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     2581 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     2641 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     2701 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     2761 cactctcggc atggacgagc tgtactagct tgtacaaagt tggcattata agaaagcatt
     2821 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc
     2881 atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc
     2941 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaata atatcatcat
     3001 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     3061 aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     3121 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc
     3181 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     3241 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     3301 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat
     3361 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     3421 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     3481 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     3541 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc
     3601 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     3661 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     3721 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     3781 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     3841 tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag agcattacgc
     3901 tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt
     3961 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4021 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4081 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4141 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4201 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4261 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4321 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4381 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4441 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4501 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4561 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4621 tcctggcctt ttgctggcct tttgctcaca tgtt
//
