LOCUS       sequence-190170-        4392 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses HA tagged syntaxin 1 in Drosophila cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4392)
  TITLE     Peden lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4392)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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            "Unpublished"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-STX1
FEATURES             Location/Qualifiers
     source          1..4392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(170..758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(250..269)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(929..1720,1721..1789))
                     /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     1552..1571
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1790..1894)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1962..1980
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2018..2040)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2140..2159
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2353..2375
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2367..2384
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2368..2384
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2390..2816
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     CDS             2848..2874
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3772..3813
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3823..3840
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3863..3880)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3882..4016
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3906..3925)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3960..3979
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4160..4176)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4160..4176)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4173..4195)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4184..4200
                     /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(4208..4214,4215..4232,4233..4238))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4253..4274
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(join(4391..4392,1..16))
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
       61 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      121 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
      181 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
      241 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
      301 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
      361 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
      421 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
      481 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
      541 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
      601 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
      661 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
      721 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
      781 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
      841 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
      901 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
      961 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1021 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     1081 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1141 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     1201 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     1261 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     1321 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     1381 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     1441 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     1501 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     1561 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     1621 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     1681 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     1741 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     1801 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     1861 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     1921 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     1981 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     2041 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     2101 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     2161 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     2221 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     2281 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     2341 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaatta attcgttgca
     2401 ggacaggatg tggtgcccga tgtgactagc tctttgctgc aggccgtcct atcctctggt
     2461 tccgataaga gacccagaac tccggccccc caccgcccac cgccaccccc atacatatgt
     2521 ggtacgcaag taagagtgcc tgcgtatgcc ccatgtgccc caccaagagc tttgcatccc
     2581 atacaagtcc ccaaagtgga gaaccgaacc aattcttcgc gggcagaaca aaagcttctg
     2641 cacacgtctc cactcgaatt tggagccggc cggcgtgtgc aaaagaggtg aatcgaacga
     2701 aagacccgtg tgtaaagccg cgtttccaaa atgtataaaa ccgagagcat ctggccaatg
     2761 tgcatcagtt gtggtcagca gcaaaatcaa gtgaatcatc tcagtgcaac taaagggggg
     2821 atctagatcg gggtacctac tagtatgtac ccctacgacg tccccgacta cgcaactaaa
     2881 gacagattag ccgctctcca tgccgcccaa tccgatgacg aggaggagac ggaggtggcc
     2941 gtcaatgtgg atggccatga ttcctacatg gacgacttct tcgcccaggt ggaggagatc
     3001 cgggggatga tcgacaaggt gcaggataac gtcgaggagg taaagaagaa gcactcggcc
     3061 atcctgtccg ccccacaaac ggacgagaag accaagcaag agctggagga tctgatggcc
     3121 gacatcaaga agaatgccaa tcgcgttcgc ggcaagctca agggcatcga gcagaacatc
     3181 gagcaggaag agcagcagaa taagtcgtcg gcggatctga gaattcggaa gacgcagcac
     3241 tcgacgttgt cacggaaatt cgtcgaagtg atgaccgagt acaatcgcac gcagaccgac
     3301 taccgagagc gctgcaaggg aaggatacag cgtcaactgg agattaccgg acgaccgacc
     3361 aacgacgatg agctggagaa gatgctggag gagggcaact cgtctgtgtt cacgcagggc
     3421 atcatcatgg agacgcagca ggccaaacag acgctggcgg acattgaggc ccgccaccag
     3481 gacatcatga agctggagac atcgatcaag gagctgcacg acatgttcat ggacatggcc
     3541 atgctggtgg agtcgcaggg cgagatgatc gatcgcattg agtaccatgt ggagcacgct
     3601 atggactatg tgcagacggc aactcaggac accaagaagg cgctcaagta ccagagtaaa
     3661 gcccgacgaa agaagatcat gatactgatc tgcctcactg tgctgggcat cttagcggcc
     3721 tcatatgtta gcagttattt catgtaactc gagtctagag ggcccttcga aggtaagcct
     3781 atccctaacc ctctcctcgg tctcgattct acgcgtaccg gtcatcatca ccatcaccat
     3841 tgagtttaaa cccgctgatc agcctcgact gtgccttcta agatccagac atgataagat
     3901 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     3961 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     4021 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     4081 gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tcagtcgacc tgcaggcatg
     4141 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4201 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4261 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4321 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4381 cttccgcttc ct
//
