LOCUS       sequence-310637-        3767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone containing HyPep presynaptic component (Hydra 
            RFamide) for HySyn.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3767)
  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 3767)
  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: pJH161_pE12_HyPep_mRuby
FEATURES             Location/Qualifiers
     source          1..3767
                     /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             1148..1858
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     protein_bind    complement(1892..1991)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2008..2027)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2009..2027)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2032..2048)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2032..2048)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2150..2169)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2161..2970
                     /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(2228..2247)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3117..3705
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3606..3625
                     /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 gccaccatgc tgggaaacaa aagaatggga ctgtgtggac tgaccctggc
      721 cctgtctctg ctggtgtgtc tgggaatttt ggctgaggga tatccatctc cagataatcc
      781 aggagaggac gccccagctg aagatgagac aggagctgaa gatctgggag aggagcacgc
      841 ttggaaggct agccatagac agaaaagaca gtggctggga ggaagattcg gaaaaagatc
      901 tggagaagag gaggatcatg atatgatgaa gagacagtgg ctcggaggaa gatttggaaa
      961 gagaagcgga gatgaggagg ccgagactag tcacagacag aagagacagt ggtttaacgg
     1021 aagattcggc aaaagatctg gcgaggagga agatcacgat atgatgaaaa gacagtggtt
     1081 caacggaaga tttggcaaaa gatccggaga tgaggaagcc tctagaggat cccacagaca
     1141 aaaaagaatg gtgtctaagg gcgaagagct gatcaaggaa aatatgcgta tgaaggtggt
     1201 catggaaggt tcggtcaacg gccaccaatt caaatgcaca ggtgaaggag aaggcaatcc
     1261 gtacatggga actcaaacca tgaggatcaa agtcatcgag ggaggacccc tgccatttgc
     1321 ctttgacatt cttgccacgt cgttcatgta tggcagccgt acttttatca agtacccgaa
     1381 aggcattcct gatttcttta aacagtcctt tcctgagggt tttacttggg aaagagttac
     1441 gagatacgaa gatggtggag tcgtcaccgt catgcaggac accagccttg aggatggctg
     1501 tctcgtttac cacgtccaag tcagaggggt aaactttccc tccaatggtc ccgtgatgca
     1561 gaagaagacc aagggttggg agcctaatac agagatgatg tatccagcag atggtggtct
     1621 gaggggatac actcatatgg cactgaaagt tgatggtggt ggccatctgt cttgctcttt
     1681 cgtaacaact tacaggtcaa aaaagaccgt cgggaacatc aagatgcccg gtatccatgc
     1741 cgttgatcac cgcctggaaa ggttagagga aagtgacaat gaaatgttcg tagtacaacg
     1801 cgaacacgca gttgccaagt tcgccgggct tggtggtggg atggacgagc tgtacaagtg
     1861 aaagagatct ggagatgaag aatgatcatg aacccagctt tcttgtacaa agttggcatt
     1921 ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa
     1981 atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct
     2041 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     2101 ataatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     2161 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     2221 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     2281 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     2341 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     2401 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     2461 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     2521 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     2581 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     2641 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     2701 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     2761 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     2821 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     2881 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     2941 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     3001 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     3061 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3121 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3181 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3241 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3301 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3361 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3421 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3481 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3541 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3601 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3661 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3721 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
