LOCUS       sequence-318829-        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     98..115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(274..301)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(393..479)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     528..550
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     542..559
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     543..559
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    575..674
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1154..1864
                     /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(1898..1997)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2014..2033)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2015..2033)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2038..2054)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2038..2054)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2156..2175)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2167..2976
                     /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(2234..2253)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3123..3711
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3612..3631
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
       61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      181 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg
      241 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
      301 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
      361 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
      421 gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga
      481 tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg
      541 ttgtaaaacg acggccagtc ttaagctcgg gccccaaata atgattttat tttgactgat
      601 agtgacctgt tcgttgcaac aaattgatga gcaatgcttt tttataatgc caactttgta
      661 caaaaaagca ggcttagcca ccatgctggg aaacaaaaga atgggactgt gtggactgac
      721 cctggccctg tctctgctgg tgtgtctggg aattttggct gagggatatc catctccaga
      781 taatccagga gaggacgccc cagctgaaga tgagacagga gctgaagatc tgggagagga
      841 gcacgcttgg aaggctagcc atagacagaa aagacagtgg ctgggaggaa gattcggaaa
      901 aagatctgga gaagaggagg atcatgatat gatgaagaga cagtggctcg gaggaagatt
      961 tggaaagaga agcggagatg aggaggccga gactagtcac agacagaaga gacagtggtt
     1021 taacggaaga ttcggcaaaa gatctggcga ggaggaagat cacgatatga tgaaaagaca
     1081 gtggttcaac ggaagatttg gcaaaagatc cggagatgag gaagcctcta gaggatccca
     1141 cagacaaaaa agaatggtgt ctaagggcga agagctgatc aaggaaaata tgcgtatgaa
     1201 ggtggtcatg gaaggttcgg tcaacggcca ccaattcaaa tgcacaggtg aaggagaagg
     1261 caatccgtac atgggaactc aaaccatgag gatcaaagtc atcgagggag gacccctgcc
     1321 atttgccttt gacattcttg ccacgtcgtt catgtatggc agccgtactt ttatcaagta
     1381 cccgaaaggc attcctgatt tctttaaaca gtcctttcct gagggtttta cttgggaaag
     1441 agttacgaga tacgaagatg gtggagtcgt caccgtcatg caggacacca gccttgagga
     1501 tggctgtctc gtttaccacg tccaagtcag aggggtaaac tttccctcca atggtcccgt
     1561 gatgcagaag aagaccaagg gttgggagcc taatacagag atgatgtatc cagcagatgg
     1621 tggtctgagg ggatacactc atatggcact gaaagttgat ggtggtggcc atctgtcttg
     1681 ctctttcgta acaacttaca ggtcaaaaaa gaccgtcggg aacatcaaga tgcccggtat
     1741 ccatgccgtt gatcaccgcc tggaaaggtt agaggaaagt gacaatgaaa tgttcgtagt
     1801 acaacgcgaa cacgcagttg ccaagttcgc cgggcttggt ggtgggatgg acgagctgta
     1861 caagtgaaag agatctggag atgaagaatg atcatgaacc cagctttctt gtacaaagtt
     1921 ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa
     1981 aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc
     2041 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     2101 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     2161 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
     2221 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     2281 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     2341 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     2401 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     2461 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     2521 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     2581 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     2641 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     2701 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     2761 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     2821 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     2881 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     2941 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
     3001 cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt
     3061 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3121 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3181 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3241 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     3301 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3361 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3421 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3481 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3541 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3601 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3661 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3721 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctca
//
