LOCUS       sequence-429974-        5907 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cal-ID (V5) for AAV production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5907)
  AUTHORS   Kim JW, Yong AJH, Aisenberg EE, Lobel JH, Wang W, Dawson TM, Dawson 
            VL, Gao R, Jan YN, Bateup HS, Ingolia NT
  TITLE     Molecular recording of calcium signals via calcium-dependent 
            proximity labeling.
  JOURNAL   Nat Chem Biol. 2024 Jul;20(7):894-905. doi: 
            10.1038/s41589-024-01603-7. Epub 2024 Apr 24.
  PUBMED    38658655
REFERENCE   2  (bases 1 to 5907)
  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"; doi: 
            "10.1038/s41589-024-01603-7"; journalName: "Nat Chem Biol"; date: 
            "2024-07"; volume: "20"; issue: "7"; pages: "894-905"
COMMENT     Sequence Label: pNTI825
FEATURES             Location/Qualifiers
     source          1..5907
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        181..628
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             714..770
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     CDS             join(771..779,780..794)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             2229..2270
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    2298..2886
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2351..2371)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2769..2780)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2928..2945)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2934..3141
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3170..3310
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3170..3299
                     /label=AAV2 ITR
     rep_origin      3385..3840
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3472..3491)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3682..3703
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3857..3876)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3976..3998
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4036..4054)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4122..4226
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4227..4295,4296..5087)
                     /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"
     rep_origin      5258..5846
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5747..5766
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgtaca gtgtctagac tgcagagggc cctgcgtatg
      181 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
      241 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      301 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      361 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      421 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      481 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      541 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      601 aggagtcgtg tcgtgcctga gagcgcagtc gagaaggatc cgggctgcag gtcgactcta
      661 gagccaccat gaacgacatc ttcgaggctc agcgtatcga atggcactca tcacgtcgta
      721 agtggaataa gacaggtcac gcagtcagag ctataggtcg gctgagctca gatctgtacg
      781 acgatgacga taagctcgag gggagcgtgg cagtcctgcc tgtggtcgac tccaccaatc
      841 agtacctgct ggatcgaatc ggcgagctga agagtgggga tgcttgcatt gcagaatatc
      901 agcaggcagg gagaggaagc agagggagga aatggttctc tccttttgga gctaacctgt
      961 acctgagtat gttttggcgc ctgaagcggg gaccagcagc aatcggcctg ggcccggtca
     1021 tcggaattgt catggcagaa gcgctgcgaa agctgggagc agacaaggtg cgagtcaaat
     1081 ggcccaatga cctgtatctg caggatagaa agctggcagg catcctggtg gagctggccg
     1141 gaataacagg cgatgctgca cagatcgtca ttggcgccgg gattaacgtg gctatgaggc
     1201 gcgtggagga aagcgtggtc aatcagggct ggatcacact gcaggaagca gggattaacc
     1261 tggacaggaa tactctggcc gctacgctga tccgagagct gcgggcagcc ctggaactgt
     1321 tcgagcagga aggcctggct ccatatctgc cacggtggga gaagctggat aacttcatca
     1381 atagacccgt gaagctgatc attggggaca aagagatttt cgggattagc cgggggattg
     1441 ataaacaggg agccctgctg ctggaacagg acggagttat caaaccctgg atgggcggag
     1501 aaatcagtct gcggtctgcc gaaaagctgc agggcggtac cggagggagc aaagacaata
     1561 ctgtgcctct gaagctgatc gctctcctgg ctaatggcga gttccatagt ggcgaacagc
     1621 tgggagaaac cctgggcatg tccagggccg ctatcaacaa gcacattcag actctgcgcg
     1681 actggggcgt ggacgtgttc accgtgcccg gaaagggcta ctctctgccc gagcctatcc
     1741 cgctgctgaa cgctaaacag attctgggac agctggacgg cctgccggac caactgactg
     1801 aagagcagat cgcagaattt aaagaggctt tctccctatt tgacaaggac ggggatggga
     1861 caataacaac caaggagctg gggacggtga tgcggtctct ggggcagaac cccacagaag
     1921 cagagctgca ggacatgatc aatgaagtag atgccgacgg tgacggcaca atcgacttcc
     1981 ctgagttcct gacaatgatg gcaagaaaaa tgaaatacag ggacacggaa gaagaaatta
     2041 gagaagcgtt cggtgtgttt gataaggatg gcaatggcta catcagtgca gcagagcttc
     2101 gccacgtgat gacaaacctt ggagagaagt taacagatga agaggttgat gaaatgatca
     2161 gggaagcaga catcgatggg gatggtcagg taaactacga agagtttgta caaatgatga
     2221 cagcgaaggg caagcccatc cccaaccccc tgctgggcct ggacagcacc tgagaattcg
     2281 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     2341 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     2401 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     2461 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     2521 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     2581 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     2641 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     2701 cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct
     2761 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     2821 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     2881 ccccgcatcg ataccgagcg ctgctcgagc tagagctcgc tgatcagcct cgactgtgcc
     2941 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3001 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3061 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3121 caatagcagg catgctgggg aggtaaccac gtgcggaccg agcggccgca ggaaccccta
     3181 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     3241 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     3301 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3361 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3421 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg
     3481 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3541 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3601 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3661 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta
     3721 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa
     3781 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     3841 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     3901 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     3961 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     4021 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     4081 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4141 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4201 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     4261 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4321 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4381 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4441 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4501 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4561 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4621 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4681 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4741 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     4801 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4861 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4921 aatctggagc cggtgagcgt ggaagccgcg gtatcattgc agcactgggg ccagatggta
     4981 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5041 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     5101 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5161 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5221 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5281 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5341 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5401 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5461 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5521 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5581 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5641 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5701 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5761 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5821 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5881 ccttttgctg gccttttgct cacatgt
//
