LOCUS       sequence-316158-        4710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone containing postsynaptic HyCal component (HyNaC2)
            for HySyn system fused to eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4710)
  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 4710)
  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: pJH198_pE12_hyCal2-eGFP
FEATURES             Location/Qualifiers
     source          1..4710
                     /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(1584..1586,1587..2294)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELY"
     primer_bind     complement(1626..1647)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1887..1906)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2234..2255
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2299..2398)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2415..2434)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2416..2434)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2439..2455)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2439..2455)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2557..2576)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2568..3377
                     /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(2635..2654)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3524..4112
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4013..4032
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4266..4283
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4442..4469)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4561..4647)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4696..4710,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4710,1..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 aggctagatt cgccagaaga atggagaaaa
      181 tcgtggagga aagaatgacc gtgaaacaga tttttaatag atacgtgcag acctctaccc
      241 tgcacggatt tcggttcatc ttcatggata ccttcattgt gagaagagtg ctgtggacaa
      301 tcctgaccct gacaatggct acaattttct ttaaggaact gagaaattcc atcaatctgt
      361 tctatgagta tccattcaca acaaccagca ccatccagta cgaaccatct ctgacattcc
      421 cagctattag cgtgtgtaat ctgaatcact ttctgctgag caaaatcaag aagagcaaac
      481 tgaagccact gtatgatcag ggaagactgc catttgacaa caatctggag aatccaggat
      541 tcgatattca gggagaggag ctgtacagca ttctgaagac aagctctcag agcattgatg
      601 aaatttttct gtcttgcgaa tggaaaagca gagacacagc caagaatgga gtgccaaatc
      661 catgcaaacc aaacaacttc acagtgtaca gcggactgta cggacagagc tgctacacat
      721 tcaatccagg agtgagcgga tatccactgc tgtctctgag cgaaaccgga gtgaacatgg
      781 gattcaagct ggagctggat ctgaagtcta aggactctct gcagggaatt caggagattg
      841 gagctatcgt gatcgtgcat caccagcagg agaccccagt gctgcaggcc ggattcgtgg
      901 tgtctccagg attccagaca ttcgtggaga ttaaggtgag acagaccgaa aatctgccac
      961 caccatatgc tacaaaatgc ggatctaaac cactgaagaa ctatcagatc tacagacagt
     1021 ctagctgctt tctggaacag ctgggagatg ccattgaaac aaagtgcaaa tgcaagagct
     1081 cttttatggc tggaagaaat atcccatact gctctctgag acagaccgtg acatgtctga
     1141 tgccaaccat ctacgacttt gacagaaaga caaataacaa ttgcccagtg gactgtgaaa
     1201 caattcagta cctgtctagc ctgtcttacg ctagattcat tagcaatgtg acatacctga
     1261 gcaagaacgc tgaacagagc agctacatta gaaaactgaa aaatagcatg tccccaaaaa
     1321 agctgcagaa gtatatcgaa gaaaatattg tggctgtgca gttcttttac caggagatga
     1381 agaaggaaaa ggtgaaacag gagccaagct acgattttta taaactgatt ggagacgtgg
     1441 gaggacagct gggactgctg ctgggagcct ctgtgctgac actggtggag tttgtggatc
     1501 tgttcatttt taccctgtac caccagctgc tgagactgag caaaaagaag tcaggtggct
     1561 caggtggtgg cggttcaggc ggaatgagta aaggagaaga acttttcacc ggggtggtgc
     1621 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     1681 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     1741 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     1801 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1861 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1921 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1981 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     2041 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     2101 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2161 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     2221 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2281 tggacgagct gtactagcac ccagctttct tgtacaaagt tggcattata agaaagcatt
     2341 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc
     2401 atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc
     2461 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     2521 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     2581 aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     2641 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc
     2701 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     2761 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     2821 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat
     2881 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     2941 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     3001 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     3061 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc
     3121 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     3181 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     3241 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     3301 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     3361 tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag agcattacgc
     3421 tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt
     3481 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3541 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3601 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3661 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3721 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3781 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3841 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3901 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3961 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4021 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4081 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4141 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4201 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4261 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     4321 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     4381 gcagtgagcg caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc
     4441 aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg
     4501 gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg
     4561 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
     4621 ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc
     4681 tagcatggat gttttcccag tcacgacgtt
//
