LOCUS       sequence-348828-        5567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the promiscuous small biotin ligase, BioID2, in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5567)
  AUTHORS   Soto JS, Jami-Alahmadi Y, Chacon J, Moye SL, Diaz-Castro B, 
            Wohlschlegel JA, Khakh BS
  TITLE     Astrocyte-neuron subproteomes and obsessive-compulsive disorder 
            mechanisms.
  JOURNAL   Nature. 2023 Apr;616(7958):764-773. doi: 10.1038/s41586-023-05927-7.
            Epub 2023 Apr 12.
  PUBMED    37046092
REFERENCE   2  (bases 1 to 5567)
  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/s41586-023-05927-7"; journalName: "Nature"; date: 
            "2023-04"; volume: "616"; issue: "7958"; pages: "764-773"
COMMENT     Sequence Label: PZac2.1 hSyn1-BioID2-BioID2-HA
FEATURES             Location/Qualifiers
     source          1..5567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   259..388
                     /label=AAV2 ITR
     promoter        465..912
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     intron          959..1091
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1074..1093
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1136..1154
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1181..1879
                     /codon_start=1
                     /product="promiscuous R40G mutant of a biotin protein 
                     ligase from Aquifex aeolicus (Kim et al., 2016)"
                     /label=BioID2
                     /translation="MFKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRKW
                     LSQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKKV
                     SGVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRISE
                     NLKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIKE
                     ILSGEFSLRRS"
     CDS             1889..1915
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    1940..2528
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1993..2013)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2411..2422)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2592..2713
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2603..2622)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2657..2676
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2875..3004
                     /label=AAV2 ITR
     primer_bind     complement(3023..3040)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3023..3039)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3032..3054)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3181..3636
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3268..3287)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3478..3499
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3662..3766
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3767..3835,3836..4627)
                     /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(3985..4004)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4798..5386
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5287..5306
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccagattta
      241 attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      301 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      361 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      421 tctacgtagc catgctctag gaagatctga gggccctgcg tatgagtgca agtgggtttt
      481 aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga cccactggac
      541 aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg gaggggaaac
      601 aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt gccttcgccc
      661 ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc
      721 ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc cgccggccca
      781 gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc
      841 ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc
      901 ctgagagcgc agtcgagaga acttaagctg cagaagttgg tcgtgaggca ctgggcaggt
      961 aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca
     1021 gagaaaactc ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt
     1081 tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga gtacttaata
     1141 cgactcacta taggctagcc tcgagctcgg atccaccacc atgttcaaaa atcttatttg
     1201 gctgaaggaa gttgatagca cccaggagcg gcttaaggaa tggaatgtct cctatggcac
     1261 tgcactcgtc gccgatagac agaccaaagg tagaggcggt ctcggacgga agtggttgag
     1321 tcaggagggc ggtctttact tctctttctt gttgaatcca aaagagtttg aaaatttgtt
     1381 gcaacttcca ctggtcttgg gactctccgt gagcgaggca ctcgaagaga ttacagagat
     1441 accatttagt ctgaagtggc ccaacgacgt ttattttcag gagaaaaagg tttcaggggt
     1501 cctttgtgag ctcagtaagg acaagctgat agtgggaatc ggtataaacg tgaatcaaag
     1561 agagatccct gaagagatca aggatcgggc cacaacactt tatgaaatca ctggaaagga
     1621 ttgggacaga aaggaggtct tgctgaaggt tctcaagcga atttccgaaa atctcaagaa
     1681 atttaaggaa aaatccttta aagagttcaa aggaaaaatt gagtcaaaga tgttgtacct
     1741 tggagaggag gttaaactct tgggagaagg caagattacc gggaaacttg tgggcctgag
     1801 tgaaaagggc ggtgctttga tacttacaga ggaaggcata aaggagatac tgtccggcga
     1861 atttagcttg cgacggtcag ctagcgctta cccttacgat gtaccggatt acgcataggg
     1921 atccaattcg cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     1981 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     2041 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     2101 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     2161 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     2221 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2281 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     2341 gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg
     2401 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     2461 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     2521 ctccccgcat cgataccgga attctgcaga tatctagagt cgacccgggc ggccgcttcg
     2581 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2641 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2701 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     2761 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct
     2821 tcctagagca tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac
     2881 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     2941 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     3001 gcagccttaa ttaacctaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     3061 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     3121 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     3181 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3241 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3301 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3361 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3421 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3481 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3541 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3601 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     3661 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3721 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3781 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3841 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3901 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3961 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4021 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     4081 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4141 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     4201 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4261 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     4321 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4381 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4441 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4501 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4561 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4621 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     4681 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4741 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4801 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4861 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4921 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4981 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5041 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5101 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5161 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5221 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5281 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5341 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5401 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5461 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     5521 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
