LOCUS       sequence-477850-        3477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with N-terminal hedgehog (hhN) tagged with HA 
            (contains stop codon).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3477)
  AUTHORS   Bosch JA, Beltran PMJ, Cavers C, LaGraff JT, Melanson R, Singh A, 
            Chen W, Hu Y, Tattikota SG, Liu Y, Hashmi Y, Asara JM, Branon T, 
            Ting AY, Carr SA, Perrimon N
  TITLE     Multi-omic mapping of Drosophila protein secretomes reveals 
            tissue-specific origins and inter-organ trafficking
  JOURNAL   bioRxiv 2025.07.09.659702; doi: 
            https://doi.org/10.1101/2025.07.09.659702
REFERENCE   2  (bases 1 to 3477)
  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: "bioRxiv 
            2025.07.09.659702; doi: https://doi.org/10.1101/2025.07.09.659702"
COMMENT     Sequence Label: pENTR_hhN-HA
FEATURES             Location/Qualifiers
     source          1..3477
                     /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             1487..1513
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1532..1579
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     protein_bind    complement(1602..1701)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1718..1737)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1719..1737)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1742..1758)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1742..1758)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1860..1879)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1871..2680
                     /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(1938..1957)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2827..3415
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3316..3335
                     /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 caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc gcggccgccc ccttcaccat ggataaccac agctcagtgc cttgggccag
      721 tgccgccagt gtcacctgtc tctccctgga tgccaaatgc cacagttcca gttccagttc
      781 cagctccaaa tccgcagcga gctccatctc cgcaatcccg caagaagaaa cgcaaacgat
      841 gcgccacatt gcgcatacgc agcgttgcct cagcaggctg acctctctgg tggccctgct
      901 gctgatcgtc ttgccgatgg tctttagccc ggctcacagc tgcggtcctg gccgaggatt
      961 gggtcgtcat agggcgcgca acctgtatcc gctggtcctc aagcagacaa ttcccaatct
     1021 atccgagtac acgaacagcg cctccggacc tctggagggt gtgatccgtc gggactcgcc
     1081 caaattcaag gacctcgtgc ccaactacaa cagggacatc cttttccgcg acgaggaagg
     1141 caccggagcg gatcgcttga tgagcaagcg ctgcaaggag aagctaaacg tgctggccta
     1201 ctcggtgatg aacgaatggc ccggcatccg gctgctggtc accgagagct gggacgagga
     1261 ctaccatcac ggccaggagt cgctccacta cgagggccga gcggtgacca ttgccacctc
     1321 cgatcgcgac cagtccaaat acggcatgct cgctcgcctg gccgtcgagg ctggattcga
     1381 ttgggtctcc tacgtcagca ggcgccacat ctactgctcc gtcaagtcag attcgtcgat
     1441 cagttcccac gtgcacggcg gtgatggtgg ctcgggtgga ggaagttacc catacgatgt
     1501 tccagattac gctggaggct ccggtactag ccgtgaccac atggtccttc atgagtatgt
     1561 aaatgctgct gggattacat aatagaaggg tgggcgcgcc gacccagctt tcttgtacaa
     1621 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     1681 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     1741 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     1801 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     1861 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     1921 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     1981 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     2041 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     2101 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     2161 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     2221 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     2281 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     2341 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     2401 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     2461 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     2521 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     2581 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     2641 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     2701 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     2761 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2821 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2881 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2941 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3001 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3061 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3121 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3181 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3241 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3301 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3361 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3421 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
