LOCUS       sequence-450307-        5683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This AAV vector contains a ChAT enhancer that drives mCherry 
            expression in spinal cord skeletal motor neurons. Specificity of 
            vector not characterized outside the mouse spinal cord..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5683)
  TITLE     AAV vector for gene delivery to spinal cord skeletal motor neurons
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5683)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pTP-AAV-ChATen-mCherry
FEATURES             Location/Qualifiers
     source          1..5683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2069..2090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2281..2300
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2459..2588
                     /label=AAV2 ITR
     promoter        3662..3693
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     intron          3694..3826
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      3844..3853
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3850..4560
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDIPSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3995..4013)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4263..4282
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    4585..5173
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4638..4658)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5056..5067)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5216..5337
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5227..5246)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5281..5300
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5499..5628
                     /label=AAV2 ITR
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
     2641 gaagagtagg tacccagtga gcttcattat cacctaacag cttcagagtg ggtggtgggt
     2701 tttggatgac aacctttctt ctcattttat tcagtggcca caccgtggcc ttagtctgat
     2761 aaaccaaaaa cctgctccat tatgaatcag tgctgtgggg agtgggtaga gagtgtgaag
     2821 ttctggggtg ggggagtctg gagagagggt gggagcagcc attctgcagc agtgccttct
     2881 tggggtcatg ggtctgtagg tgctgctgtg gagggagaga tcagcctatt ctggcttcat
     2941 ttctgagctg caaactgcct gggtgtctgg agaagcaggt tggcgtggtg gttagcagtg
     3001 cgtgggcggg gttgcccgct cttgatttat gatttctttg tctctgtgga agcacttaag
     3061 tgcaggcttt agttccaatg acactcagga gcctctggat tccagcactg gggatggggg
     3121 tggggtagaa cgttctcagg cctcaccaac ccctcccctg tgtgctgcct ttgggagagt
     3181 cccaaggctt cagcattact taattaatta ggcctctact gctacatagg ctcagattca
     3241 aaagaacaga gtggcccacg tcagccattc ccggaaaagt ctgatggctg gaagccagag
     3301 gactatgtgt ctgccttgct gcccttggcc agcccatcct gaatgcccag actcggacaa
     3361 tggagtaggt acagaagggt aaagacagtg tcttctgtac cagtaagtgg gccctgatct
     3421 gctctctaca gcttccagag aaagggcctg gccaatgagc ggccttttga gtagcagata
     3481 cctcacatgc attctgatag aaagcctggc cccagatcac tgtgacttta gccctcaggt
     3541 ttcttttgca cttcaattca atgacttctt gaggttcatt tccctctcca agatttgcca
     3601 cagaccagtg gttctcaacc tgtgggtcac acctcctttg gggaaattga atgaaccggt
     3661 ctagagggta tataatggaa gctcgacttc caggtaagta tcaaggttac aagacaggtt
     3721 taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag
     3781 gcacctattg gtcttactga catccacttt gcctttctct ccacagggca aagaattgga
     3841 tccgccacca tggtgagcaa gggcgaggag gacaacatgg ccatcatcaa ggagttcatg
     3901 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     3961 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc
     4021 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     4081 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
     4141 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     4201 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
     4261 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc
     4321 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
     4381 tacgacgccg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     4441 tacaacgtca acatcaagct ggacatcccc tcccacaacg aggactacac catcgtggaa
     4501 cagtacgagc gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa
     4561 gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     4621 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     4681 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     4741 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     4801 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     4861 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     4921 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     4981 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     5041 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     5101 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     5161 gccgcctccc cgcatcgata ccgtcgaccc gggcggccgc ttcgagcaga catgataaga
     5221 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     5281 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     5341 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     5401 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag agcatggcta
     5461 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
     5521 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     5581 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcattaatga
     5641 atcggccaac gcgcggggag aggcggtttg cgtattgggc gct
//
