LOCUS       sequence-487697-        6249 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express Fas-EGFP and mNaChBac in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6249)
  AUTHORS   Li H, Su C, Xu Y, Ludwig MQ, Davis J, Tong Q
  TITLE     An alternative neural basis underlying leptin resistance.
  JOURNAL   Cell Rep. 2025 Jun 19;44(7):115863. doi: 
            10.1016/j.celrep.2025.115863.
  PUBMED    40540394
REFERENCE   2  (bases 1 to 6249)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep.";
            date: "2025-06-19"; volume: "44(7):115863. doi"; pages: " 
            10.1016/j.celrep.2025.115863"
COMMENT     Sequence Label: pAAV-SYN1>Fas-EGFP:P2A:mNaChBac.
FEATURES             Location/Qualifiers
     source          1..6249
                     /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        166..613
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      686..695
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(692..694,695..697,698..1408)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(692..694,695..697,698..1408)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(737..758)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(998..1017)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1345..1366
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1424..1480
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     misc_feature    2376..2964
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2429..2449)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2847..2858)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2996..3472
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3195..3214)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3512..3652
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3512..3641
                     /label=AAV2 ITR
     rep_origin      3727..4182
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3814..3833)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4024..4045
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4199..4218)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4318..4340
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4378..4396)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4464..4568
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4569..4637,4638..5429)
                     /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(4787..4806)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5600..6188
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6089..6108
                     /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 gcgtctgcag agggccctgc gtatgagtgc aagtgggttt
      181 taggaccagg atgaggcggg gtgggggtgc ctacctgacg accgaccccg acccactgga
      241 caagcaccca acccccattc cccaaattgc gcatccccta tcagagaggg ggaggggaaa
      301 caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag tgccttcgcc
      361 cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga cgtcactcgc
      421 cggtcccccg caaactcccc ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc
      481 agccggaccg caccacgcga ggcgcgagat aggggggcac gggcgcgacc atctgcgctg
      541 cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg
      601 cctgagagcg cagggtaccg gatcctctag agtcgactcc ggaacaactt cgtatatacc
      661 tttctatacg aagttgtggc gcgccgccac catggtgagc aagggcgagg agctgttcac
      721 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
      781 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
      841 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
      901 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
      961 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     1021 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     1081 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     1141 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     1201 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     1261 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     1321 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     1381 cactctcggc atggacgagc tgtacaagct cgagggaagc ggagccacta acttctccct
     1441 gttgaaacaa gcaggggatg tcgaagagaa tcccgggcca tctagaatga aaatggaagc
     1501 acgccagaaa cagaactcat tcacgtcaaa gatgcagaaa attgtcaacc atagagcgtt
     1561 tactttcacc gtcattgccc tgatcctctt taatgcactg atcgtgggaa tcgaaacata
     1621 tccgaggatc tacgcggacc acaagtggct cttttatcgg attgacctcg tcctgctttg
     1681 gatcttcacc atcgagatcg cgatgaggtt tttggcgtcg aacccaaagt cggccttctt
     1741 ccgcagctcg tggaattggt ttgattttct gattgtcgcg gctgggcata tcttcgcggg
     1801 agcccagttc gtgacagtat tgcgaatcct tagagtgctg cgggtactca gggcgatctc
     1861 cgtcgtcccc tccctgcgca gactcgtaga tgcgctggtg atgacgattc cggcgttggg
     1921 caatatcttg atcttgatgt ccatcttctt ctatatcttt gcagtgatcg ggacaatgct
     1981 cttccagcac gtatcgcctg aatactttgg gaacctccag ctttcgttgc tgacgctgtt
     2041 ccaagtggtg accttggaga gctgggccag cggtgtaatg cggcccattt ttgcagaggt
     2101 gccctggtca tggctctact ttgtctcgtt tgtcctgatt ggtactttta tcattttcaa
     2161 tcttttcatc ggagtcattg tgaacaacgt ggaaaaggcg gagcttacgg acaatgagga
     2221 agatggggag gccgacggac ttaaacagga gatttcagca ttgcgaaagg atgtagccga
     2281 actgaagagc ttgctcaaac aacttaagta agctagcaca acttcgtata tacctttcta
     2341 tacgaagttg tgaattcgat atcaagctta tcgataatca acctctggat tacaaaattt
     2401 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     2461 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     2521 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     2581 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     2641 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     2701 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     2761 tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc
     2821 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     2881 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     2941 tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg
     3001 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
     3061 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
     3121 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
     3181 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
     3241 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
     3301 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
     3361 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
     3421 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt
     3481 gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     3541 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     3601 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     3661 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     3721 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3781 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     3841 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3901 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     3961 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4021 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     4081 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     4141 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     4201 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     4261 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     4321 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     4381 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     4441 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4501 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4561 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4621 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4681 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4741 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4801 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4861 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4921 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4981 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5041 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5101 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5161 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5221 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5281 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5341 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5401 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5461 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5521 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5581 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5641 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5701 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5761 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5821 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5881 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5941 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6001 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6061 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6121 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6181 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6241 ctcacatgt
//
