LOCUS       sequence-389098-        6648 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Encodes full-length wild type synaptotagmin 1 (SYT1) with an 
            N-terminal HaloTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6648)
  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 6648)
  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: "Elife."; 
            date: "2023-02-2"; volume: "12:e82568. doi"; pages: " 
            10.7554/eLife.82568"
COMMENT     Sequence Label: pEF HaloTag-SYT1
FEATURES             Location/Qualifiers
     source          1..6648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(146..250)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        312..1494
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          543..1485
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1442..1462
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1526..1535
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1532..1621
                     /codon_start=1
                     /product="signal sequence from bovine prolactin"
                     /label=prolactin signal sequence
                     /translation="MDSKGSSQKGSRLLLLLVVSNLLLCQGVVS"
     CDS             1646..2530
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     CDS             2549..2569
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(3946..3965)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4011..4066
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4065..4084)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4427..4443)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4427..4443)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4440..4462)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4451..4467
                     /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(4475..4481,4482..4499,4500..4505))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4520..4541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4600..4795
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4646..4781
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4801..4935
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4838..4857)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4892..4911
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5003..5020)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5174..5762)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5254..5273)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5933..6648,1..76,77..145))
                     /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     6556..6575
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
       61 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
      121 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
      181 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
      241 gggttccgcg cacatttccc cgaaaagtgc cacctgggtc gacattgatt attgactaga
      301 tcatcgcgtg aggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
      361 agaagttggg gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa
      421 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
      481 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
      541 aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg
      601 tgccttgaat tacttccacg cccctggctg cagtacgtga ttcttgatcc cgagcttcgg
      661 gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct
      721 tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc
      781 gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg
      841 acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt
      901 tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg
      961 aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg
     1021 cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg
     1081 gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg
     1141 agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg
     1201 aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg
     1261 tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag
     1321 gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct
     1381 tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt
     1441 ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgaggaatt
     1501 ctgcagtcga cggtaccgcg ggcccgccac catggacagc aaaggttcgt cgcagaaagg
     1561 gtcccgcctg ctcctgctgc tggtggtgtc aaatctactc ttgtgccagg gtgtggtctc
     1621 cgatatcggt accacgcgtg gatccgaaat cggtactggc tttccattcg acccccatta
     1681 tgtggaagtc ctgggcgagc gcatgcacta cgtcgatgtt ggtccgcgcg atggcacccc
     1741 tgtgctgttc ctgcacggta acccgacctc ctcctacgtg tggcgcaaca tcatcccgca
     1801 tgttgcaccg acccatcgct gcattgctcc agacctgatc ggtatgggca aatccgacaa
     1861 accagacctg ggttatttct tcgacgacca cgtccgcttc atggatgcct tcatcgaagc
     1921 cctgggtctg gaagaggtcg tcctggtcat tcacgactgg ggctccgctc tgggtttcca
     1981 ctgggccaag cgcaatccag agcgcgtcaa aggtattgca tttatggagt tcatccgccc
     2041 tatcccgacc tgggacgaat ggccagaatt tgcccgcgag accttccagg ccttccgcac
     2101 caccgacgtc ggccgcaagc tgatcatcga tcagaacgtt tttatcgagg gtacgctgcc
     2161 gatgggtgtc gtccgcccgc tgactgaagt cgagatggac cattaccgcg agccgttcct
     2221 gaatcctgtt gaccgcgagc cactgtggcg cttcccaaac gagctgccaa tcgccggtga
     2281 gccagcgaac atcgtcgcgc tggtcgaaga atacatggac tggctgcacc agtcccctgt
     2341 cccgaagctg ctgttctggg gcaccccagg cgttctgatc ccaccggccg aagccgctcg
     2401 cctggccaaa agcctgccta actgcaaggc tgtggacatc ggcccgggtc tgaatctgct
     2461 gcaagaagac aacccggacc tgatcggcag cgagatcgcg cgctggctgt ctactctgga
     2521 gatttccggt ggatccggaa gttcggggga ggatctgtac tttcagagcg attccagcgg
     2581 ttcttcaggc agttccatgg tgagtgccag tcatcctgag gccctggccg cccctgtcac
     2641 cactgttgcg acccttgtcc cacacaatgc cactgagcca gccagtcctg gggaagggaa
     2701 ggaagatgcc ttttccaagc tgaagcagaa gtttatgaat gagctgcata aaattccatt
     2761 gccaccgtgg gccttaatag ccatagccat agttgcggtc cttttagtcg taacctgctg
     2821 cttttgtgtc tgtaagaaat gtttgttcaa aaagaaaaac aagaagaagg ggaaggaaaa
     2881 gggaggaaag aacgccatta acatgaaaga cgtgaaagac ttagggaaga ccatgaagga
     2941 tcaggccctt aaggatgacg atgctgaaac cggactgact gatggagaag aaaaggaaga
     3001 gcccaaggaa gaggagaaac tgggaaagct ccaatattca ctggactatg acttccagaa
     3061 taaccagctg ttggtgggaa tcatccaggc tgctgaactg cccgccctgg acatgggggg
     3121 tacatccgat ccatacgtca aagtcttcct gctgcctgac aaaaagaaga aatttgagac
     3181 taaagtccac cggaaaaccc tcaatccagt cttcaatgaa caatttactt tcaaggtacc
     3241 ctactcggaa ttaggtggca aaaccctggt gatggctgtg tatgactttg atcgcttctc
     3301 caagcacgac atcatcggag agttcaaagt tcctatgaac accgtggatt ttggccatgt
     3361 gaccgaggag tggcgcgatc tccagagcgc tgagaaagaa gagcaagaga aactgggtga
     3421 catctgcttc tccctccgct acgtccctac tgccggcaaa ctgactgttg tcattctgga
     3481 agccaagaac ctgaagaaga tggatgtggg tggcttatct gatccctacg tgaagattca
     3541 cctgatgcag aacggtaaga ggctgaagaa gaaaaagacg acgattaaga agaacacact
     3601 caacccctac tacaacgagt ccttcagctt tgaagttccg ttcgagcaaa tccagaaagt
     3661 gcaagtggtg gtaactgttt tggactatga caagattggc aagaacgacg ccatcggcaa
     3721 agtcttcgtt ggttacaaca gcactggggc ggagctgcga cactggtcag acatgctggc
     3781 caacccccgg cgacccatcg cacagtggca cactctgcag gtagaggagg aggtcgatgc
     3841 catgctggct gtcaagaagt aaggccgcac tcctcaggtg caggctgcct atcagaaggt
     3901 ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc
     3961 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
     4021 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     4081 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
     4141 tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa
     4201 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
     4261 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
     4321 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
     4381 cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc
     4441 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4501 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4561 ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac
     4621 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4681 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     4741 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4801 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4861 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4921 tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4981 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5041 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5101 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5161 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5221 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5281 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5341 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5401 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5461 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5521 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5581 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5641 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5701 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5761 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5821 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5881 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5941 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6001 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6061 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6121 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6181 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6241 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6301 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6361 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6421 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6481 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6541 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6601 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttga
//
