LOCUS       sequence-190169-        4518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA tagged syntaxin 4 in Drosophila cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4518)
  TITLE     Peden lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4518)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pMT-HA-STX4
FEATURES             Location/Qualifiers
     source          1..4518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     52..71
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     265..287
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     279..296
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     280..296
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        302..728
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     CDS             760..786
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1810..1851
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1861..1878
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1901..1918)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1920..2054
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1944..1963)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1998..2017
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2198..2214)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2198..2214)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2211..2233)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2222..2238
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2246..2252,2253..2270,2271..2276))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2291..2312
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2429..2446)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2600..3188)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2680..2699)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3359..4150,4151..4219))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3982..4001
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4220..4324)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4392..4410
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4448..4470)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
       61 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
      121 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
      181 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
      241 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
      301 taattcgttg caggacagga tgtggtgccc gatgtgacta gctctttgct gcaggccgtc
      361 ctatcctctg gttccgataa gagacccaga actccggccc cccaccgccc accgccaccc
      421 ccatacatat gtggtacgca agtaagagtg cctgcgcatg ccccatgtgc cccaccaaga
      481 gctttgcatc ccatacaagt ccccaaagtg gagaaccgaa ccaattcttc gcgggcagaa
      541 caaaagcttc tgcacacgtc tccactcgaa tttggagccg gccggcgtgt gcaaaagagg
      601 tgaatcgaac gaaagacccg tgtgtaaagc cgcgtttcca aaatgtataa aaccgagagc
      661 atctggccaa tgtgcatcag ttgtggtcag cagcaaaatc aagtgaatca tctcagtgca
      721 actaaagggg ggatctagat cggggtacct actagtatgt acccatacga tgttccagat
      781 tacgctggta aagatcgact gcccgagctt ttgcagcgct cattgagcac gaactcgtca
      841 aactcgtcgt cgaacggttc actgcttttg aacgtgtaca gtggaaccac ggagttcatc
      901 attggcaaca ccggcggtaa caataatagc tacagtgtcg ttagccaaaa ctctcacagc
      961 tgcagcaaca ataacagcag caccgaaccc aaggatcggt ccagttcaaa gatgacgcag
     1021 tatggctcga atgtggatga catactcaat ccgtacacgg agattcgcca acaactcgca
     1081 cagatagccg ccaacttgga gacgatgaac cgcatggccc aaactgtcaa tctgcgaact
     1141 ttcaacgaga acgagatgga tgagcttcac aacaaaaacc tgaggctggg caatcagcta
     1201 atgacgcgat tcaacgactt caaggcgaat ctgccggcgg aaaacgatta cagcttggag
     1261 gcaaggatga aaagaaccct tttctatgga ctccaccaga ctttcatcaa tctgtggcac
     1321 aaaaacgaac tattcctgca gaactacgag accaaggtca aaaaaaatct gagactgcat
     1381 acaaaaatca ttaattctga agccagcgaa caagaaatcg agttactcat cgagaacaag
     1441 acaaccaaac tctttgtgga taattttctg caggaaacgg aaaaggagcg gcagacactg
     1501 cgcgaaatga tggatcgatt caacgagctg cgccgcctgg agaagtccat cgaggaggtg
     1561 cacgccctgt tcatgcgcat ccagaccctg gtaatggagc agagcgaggt gatccagcgg
     1621 gtggagtttc acgcccagca ggccacgctc cacgtggaca agggcgccga cgagctggac
     1681 caggcggagc agcaccagaa gaaggcgcgc aagaaaaaga taatgctcat cgtgatactc
     1741 gcagccgtgt tgctagtatt actccttgtt ggtatttatt tgtgactcga gtctagaggg
     1801 cccttcgaag gtaagcctat ccctaaccct ctcctcggtc tcgattctac gcgtaccggt
     1861 catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctaag
     1921 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     1981 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     2041 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     2101 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc
     2161 agtcgacctg caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     2221 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     2281 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     2341 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2401 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2461 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2521 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2581 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2641 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2701 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2761 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2821 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2881 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2941 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3001 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3061 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3121 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3181 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3241 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3301 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3361 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3421 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3481 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3541 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3601 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3661 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3721 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3781 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3841 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3901 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3961 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4021 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4081 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4141 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4201 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4261 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4321 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     4381 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     4441 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     4501 ccgggagcag acaagccc
//
