LOCUS       sequence-389099-        6546 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes full-length wild type synaptotagmin 1 (SYT1) with a 
            C-terminal HaloTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6546)
  AUTHORS   Watson ET, Pauers MM, Seibert MJ, Vevea JD, Chapman ER
  TITLE     Synaptic vesicle proteins are selectively delivered to axons in 
            mammalian neurons.
  JOURNAL   Elife. 2023 Feb 2;12:e82568. doi: 10.7554/eLife.82568.
  PUBMED    36729040
REFERENCE   2  (bases 1 to 6546)
  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: "Elife."; 
            date: "2023-02-2"; volume: "12:e82568. doi"; pages: " 
            10.7554/eLife.82568"
COMMENT     Sequence Label: pEF SYT1-HaloTag
FEATURES             Location/Qualifiers
     source          1..6546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     494..513
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(732..836)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        898..2080
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1129..2071
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2028..2048
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      2112..2121
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3423..3443
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             3459..4343
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     primer_bind     complement(4430..4449)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4495..4550
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4549..4568)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4911..4927)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4911..4927)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4924..4946)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4935..4951
                     /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(4959..4965,4966..4983,4984..4989))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5004..5025
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5084..5279
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5130..5265
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5285..5419
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5322..5341)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5376..5395
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5487..5504)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5658..6246)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5738..5757)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6417..6546,1..662,663..731))
                     /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"
ORIGIN
        1 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
       61 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      121 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      181 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      241 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      301 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      361 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      421 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      481 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      541 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      601 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      661 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
      721 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
      781 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
      841 tttccccgaa aagtgccacc tgggtcgaca ttgattattg actagatcat cgcgtgaggc
      901 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
      961 ggggtcggca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
     1021 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
     1081 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg
     1141 tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact
     1201 tccacgcccc tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg
     1261 ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg
     1321 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc
     1381 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg
     1441 gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc
     1501 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg
     1561 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc
     1621 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc
     1681 agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag
     1741 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc
     1801 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga
     1861 ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
     1921 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
     1981 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
     2041 agtggttcaa agtttttttc ttccatttca ggtgtcgtga ggaattctgc agtcgacggt
     2101 accgcgggcc cgccaccatg gtgagtgcca gtcatcctga ggccctggcc gcccctgtca
     2161 ccactgttgc gacccttgtc ccacacaatg ccactgagcc agccagtcct ggggaaggga
     2221 aggaagatgc cttttccaag ctgaagcaga agtttatgaa tgagctgcat aaaattccat
     2281 tgccaccgtg ggccttaata gccatagcca tagttgcggt ccttttagtc gtaacctgct
     2341 gcttttgtgt ctgtaagaaa tgtttgttca aaaagaaaaa caagaagaag gggaaggaaa
     2401 agggaggaaa gaacgccatt aacatgaaag acgtgaaaga cttagggaag accatgaagg
     2461 atcaggccct taaggatgac gatgctgaaa ccggactgac tgatggagaa gaaaaggaag
     2521 agcccaagga agaggagaaa ctgggaaagc tccaatattc actggactat gacttccaga
     2581 ataaccagct gttggtggga atcatccagg ctgctgaact gcccgccctg gacatggggg
     2641 gtacatccga tccatacgtc aaagtcttcc tgctgcctga caaaaagaag aaatttgaga
     2701 ctaaagtcca ccggaaaacc ctcaatccag tcttcaatga acaatttact ttcaaggtac
     2761 cctactcgga attaggtggc aaaaccctgg tgatggctgt gtatgacttt gatcgcttct
     2821 ccaagcacga catcatcgga gagttcaaag ttcctatgaa caccgtggat tttggccatg
     2881 tgaccgagga gtggcgcgat ctccagagcg ctgagaaaga agagcaagag aaactgggtg
     2941 acatctgctt ctccctccgc tacgtcccta ctgccggcaa actgactgtt gtcattctgg
     3001 aagccaagaa cctgaagaag atggatgtgg gtggcttatc tgatccctac gtgaagattc
     3061 acctgatgca gaacggtaag aggctgaaga agaaaaagac gacgattaag aagaacacac
     3121 tcaaccccta ctacaacgag tccttcagct ttgaagttcc gttcgagcaa atccagaaag
     3181 tgcaagtggt ggtaactgtt ttggactatg acaagattgg caagaacgac gccatcggca
     3241 aagtcttcgt tggttacaac agcactgggg cggagctgcg acactggtca gacatgctgg
     3301 ccaacccccg gcgacccatc gcacagtggc acactctgca ggtagaggag gaggtcgatg
     3361 ccatgctggc tgtcaagaag ggatccggaa gttcggggtc cagcggttct tcaggcagtt
     3421 ccgaggatct gtactttcag agcgataacg atggatccga aatcggtact ggctttccat
     3481 tcgaccccca ttatgtggaa gtcctgggcg agcgcatgca ctacgtcgat gttggtccgc
     3541 gcgatggcac ccctgtgctg ttcctgcacg gtaacccgac ctcctcctac gtgtggcgca
     3601 acatcatccc gcatgttgca ccgacccatc gctgcattgc tccagacctg atcggtatgg
     3661 gcaaatccga caaaccagac ctgggttatt tcttcgacga ccacgtccgc ttcatggatg
     3721 ccttcatcga agccctgggt ctggaagagg tcgtcctggt cattcacgac tggggctccg
     3781 ctctgggttt ccactgggcc aagcgcaatc cagagcgcgt caaaggtatt gcatttatgg
     3841 agttcatccg ccctatcccg acctgggacg aatggccaga atttgcccgc gagaccttcc
     3901 aggccttccg caccaccgac gtcggccgca agctgatcat cgatcagaac gtttttatcg
     3961 agggtacgct gccgatgggt gtcgtccgcc cgctgactga agtcgagatg gaccattacc
     4021 gcgagccgtt cctgaatcct gttgaccgcg agccactgtg gcgcttccca aacgagctgc
     4081 caatcgccgg tgagccagcg aacatcgtcg cgctggtcga agaatacatg gactggctgc
     4141 accagtcccc tgtcccgaag ctgctgttct ggggcacccc aggcgttctg atcccaccgg
     4201 ccgaagccgc tcgcctggcc aaaagcctgc ctaactgcaa ggctgtggac atcggcccgg
     4261 gtctgaatct gctgcaagaa gacaacccgg acctgatcgg cagcgagatc gcgcgctggc
     4321 tgtctactct ggagatttcc ggttaaggcc gcactcctca ggtgcaggct gcctatcaga
     4381 aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct
     4441 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa
     4501 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     4561 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
     4621 catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata
     4681 tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga
     4741 ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac
     4801 atgttttact agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc
     4861 ttctcttatg gagatccctc gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg
     4921 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4981 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5041 ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aattagtcag
     5101 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5161 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     5221 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     5281 agctaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5341 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5401 atcttatcat gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5461 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5521 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5581 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5641 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5701 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5761 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5821 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5881 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5941 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6001 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6061 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6121 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6181 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6241 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6301 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6361 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6421 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6481 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6541 gctgca
//
