LOCUS       sequence-348673-        5668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP on the plasma membrane of neurons via an Lck 
            localization sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5668)
  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 5668)
  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-Lck-GFP
FEATURES             Location/Qualifiers
     source          1..5668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    4..125
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(15..34)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     69..88
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   287..416
                     /label=AAV2 ITR
     primer_bind     complement(435..452)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(435..451)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(444..466)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      593..1048
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(680..699)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     890..911
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1074..1178
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1179..1247,1248..2039)
                     /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(1397..1416)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2210..2798
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2699..2718
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    3086..3107
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3122..3127,3128..3145,3146..3152)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3160..3176
                     /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     3165..3187
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3184..3200
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3184..3200
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   3238..3367
                     /label=AAV2 ITR
     promoter        3443..3890
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     intron          3959..4091
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4074..4093
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        4136..4154
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4282..4291
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4288..4290,4291..4293,4294..5007)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4288..4290,4291..4293,4294..5004)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4333..4354)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4594..4613)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4941..4962
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5020..5608
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5073..5093)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5491..5502)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
       61 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
      121 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg
      181 ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat cttcctagag
      241 catggctacg tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg
      301 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
      361 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt
      421 aattaaccta attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
      481 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
      541 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
      601 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
      661 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
      721 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
      781 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
      841 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
      901 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
      961 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     1021 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     1081 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1141 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     1201 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     1261 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     1321 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     1381 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     1441 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     1501 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     1561 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     1621 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     1681 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     1741 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     1801 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     1861 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     1921 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     1981 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     2041 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     2101 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     2161 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2221 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2281 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2341 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2401 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2461 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2521 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2581 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2641 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2701 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2761 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2821 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     2881 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     2941 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     3001 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     3061 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     3121 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     3181 acacaggaaa cagctatgac catgattacg ccagatttaa ttaaggcctt aattaggctg
     3241 cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt
     3301 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
     3361 ggttccttgt agttaatgat taacccgcca tgctacttat ctacgtagcc atgctctagg
     3421 aagatcgagg gccctgcgta tgagtgcaag tgggttttag gaccaggatg aggcggggtg
     3481 ggggtgccta cctgacgacc gaccccgacc cactggacaa gcacccaacc cccattcccc
     3541 aaattgcgca tcccctatca gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg
     3601 cactgccagc ttcagcaccg cggacagtgc cttcgccccc gcctggcggc gcgcgccacc
     3661 gccgcctcag cactgaaggc gcgctgacgt cactcgccgg tcccccgcaa actccccttc
     3721 ccggccacct tggtcgcgtc cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc
     3781 gcgagatagg ggggcacggg cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc
     3841 ctcagtctgc ggtgggcagc ggaggagtcg tgtcgtgcct gagagcgcag tcgagagtgc
     3901 ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca ctgggcaggt
     3961 aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca
     4021 gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt
     4081 tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga gtacttaata
     4141 cgactcacta taggctagcc tcgagaattc accatgggct gtggctgcag ctcaaaccct
     4201 gaagatgact ggatggagaa cattgacgtg tgtgagaact gccattatcc cccgccgaag
     4261 cttcgaattg atccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     4321 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     4381 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     4441 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     4501 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     4561 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     4621 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     4681 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     4741 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     4801 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     4861 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     4921 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     4981 ctcggcatgg acgagctgta caagtaatct agaatcgata atcaacctct ggattacaaa
     5041 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     5101 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     5161 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     5221 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     5281 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     5341 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     5401 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt
     5461 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     5521 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     5581 cggatctccc tttgggccgc ctccccgcat cgataccgga attctgcaga tatctagagt
     5641 cgacccgggc ggccgcttcg agcagaca
//
