LOCUS       sequence-369068-        6496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the rheological probe PfV-HaloTag-SNAP-tag 
            (GEM-HaloTag-SNAP-tag) under control of human synapsin promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6496)
  AUTHORS   Wong VC, Houlihan PR, Liu H, Walpita D, DeSantis MC, Liu Z, O'Shea 
            EK
  TITLE     Plasticity-induced actin polymerization in the dendritic shaft 
            regulates intracellular AMPA receptor trafficking.
  JOURNAL   Elife. 2024 Aug 15;13:e80622. doi: 10.7554/eLife.80622.
  PUBMED    39146380
REFERENCE   2  (bases 1 to 6496)
  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: "2024-08-15"; volume: "13:e80622. doi"; pages: " 
            10.7554/eLife.80622"
COMMENT     Sequence Label: hSyn-GEM
FEATURES             Location/Qualifiers
     source          1..6496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        186..633
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             1733..2617
                     /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             2633..3175
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="DKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVEV
                     PAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLKV
                     VKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLAV
                     KEWLLAHEGHRLGKPGLG"
     polyA_signal    3242..3718
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3441..3460)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3758..3898
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3758..3887
                     /label=AAV2 ITR
     rep_origin      3973..4428
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4060..4079)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4270..4291
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4445..4464)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4564..4586
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4624..4642)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4710..4814
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4815..4883,4884..5675)
                     /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     complement(5033..5052)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5846..6434
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6335..6354
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6496,1..129)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg
       61 gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta
      121 ggggttcctg cggccgcacg cgtgagggcc tagctagctc tagactgcag agggccctgc
      181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      601 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa ggtaccgcca ccatgctctc
      661 aataaatcca acccttataa atagggataa gccatacacc aaagaagaat taatggaaat
      721 tctgagatta gcaataatag ctgagcttga tgcaataaat ctatatgagc agatggcaag
      781 atattccgaa gatgaaaacg taagaaagat tctccttgat gttgcaaggg aggagaaggc
      841 tcacgttgga gagttcatgg cactccttct aaatttggac ccagaacaag taactgagct
      901 gaagggtgga tttgaggaag tcaaggagct aactggaatt gaggctcaca taaatgataa
      961 caagaaggag gaatctaacg tggaatattt tgagaagttg agaagtgctc ttttggatgg
     1021 ggtaaataag ggaagatctt tactaaagca cttgccagta acgagaatag aaggacaatc
     1081 attcagggtt gacatcatta aatttgaaga tggtgttagg gtagtaaagc aagagtacaa
     1141 gccaattcca ttattgaaga agaaattcta tgtgggaata agagaactca acgatgggac
     1201 ttacgatgta tctatagcaa ctaaagctgg ggaactccta gttaaagacg aagagtcgct
     1261 agtaataaga gaaattctat ccacagaggg aattaagaaa atgaaacttt cctcctggga
     1321 caatccagaa gaagcactta atgatttaat gaacgcactg caggaagcct caaatgcgtc
     1381 agctggacca tttgggctga taattaaccc aaaaagatat gcaaagttgc taaaaattta
     1441 cgagaagagt ggaaagatgc tagttgaagt gctaaaagag atattcaggg gaggaatcat
     1501 tgttactcta aacattgatg agaataaggt aataatattt gccaatacac cagctgttct
     1561 tgacgttgta gttggccaag atgtgactct acaagagcta ggacctgaag gagacgacgt
     1621 tgcattcttg gttagtgaag ctataggaat aagaataaag aacccagagg caatagtagt
     1681 cttagagtgg tctggcagcg gtagtgggtc gggatcaggc tccggtggat ccgaaatcgg
     1741 tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt
     1801 cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc
     1861 ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga
     1921 cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt
     1981 ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca
     2041 cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg
     2101 tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc
     2161 ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca
     2221 gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga
     2281 gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt
     2341 cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata
     2401 catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt
     2461 tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt
     2521 ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga
     2581 gatcgcgcgc tggctgtcta ctctggagat ttccggtgaa ttcggcggaa gcgacaaaga
     2641 ctgcgaaatg aagcgcacca ccctggatag ccctctgggc aagctggaac tgtctgggtg
     2701 cgaacagggc ctgcaccgta tcatcttcct gggcaaagga acatctgccg ccgacgccgt
     2761 ggaagtgcct gccccagccg ccgtgctggg cggaccagag ccactgatgc aggccaccgc
     2821 ctggctcaac gcctactttc accagcctga ggccatcgag gagttccctg tgccagccct
     2881 gcaccaccca gtgttccagc aggagagctt tacccgccag gtgctgtgga aactgctgaa
     2941 agtggtgaag ttcggagagg tcatcagcta cagccacctg gccgccctgg ccggcaatcc
     3001 cgccgccacc gccgccgtga aaaccgccct gagcggaaat cccgtgccca ttctgatccc
     3061 ctgccaccgg gtggtgcagg gcgacctgga cgtggggggc tacgagggcg ggctcgccgt
     3121 gaaagagtgg ctgctggccc acgagggcca cagactgggc aagcctgggc tgggttaatg
     3181 agactaccgc gagaattcga tatcaagctt atcgataccg agcgctgctc gagagatcta
     3241 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     3301 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     3361 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     3421 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     3481 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     3541 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     3601 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     3661 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     3721 gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     3781 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     3841 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     3901 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     3961 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4021 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     4081 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4141 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     4201 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4261 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     4321 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     4381 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     4441 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     4501 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     4561 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     4621 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     4681 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4741 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4801 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4861 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4921 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4981 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5041 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5101 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5161 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5221 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5281 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5341 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5401 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5461 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5521 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5581 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5641 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5701 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5761 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5821 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5881 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5941 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6001 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6061 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6121 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6181 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6241 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6301 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6361 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6421 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6481 cttttgctca catgtc
//
