LOCUS       sequence-412577-        6576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Induces NM-like pigment accumulation in catecholaminergic neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6576)
  TITLE     Prigge Lab AAV vectors
REFERENCE   2  (bases 1 to 6576)
  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"
COMMENT     Sequence Label: pAAV_Ef1a_DIO_hTyrHA_minBack
FEATURES             Location/Qualifiers
     source          1..6576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      66..654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     555..574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   716..845
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        946..2124
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1177..2115
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2072..2092
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(2155..2188)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(2239..2272)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2284..2310)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    3936..3969
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    4020..4053
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4066..4469
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     polyA_signal    4476..4524
                     /note="synthetic polyadenylation signal"
     repeat_region   4554..4694
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4554..4683
                     /label=AAV2 ITR
     rep_origin      4769..5224
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4856..4875)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5066..5087
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5241..5260)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5360..5382
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5420..5438)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5506..5610
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5611..5679,5680..6471)
                     /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(5829..5848)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
       61 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
      121 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
      181 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
      241 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
      301 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
      361 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
      421 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
      481 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
      541 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
      601 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
      661 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtcctgc
      721 aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga cctttggtcg
      781 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      841 ttcctgcggc cgcacgcgta agctttgcaa agatggataa agttttaaac agagaggaat
      901 ctttgcagct aatggacctt ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt
      961 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     1021 tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     1081 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     1141 gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc
     1201 gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg
     1261 cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct
     1321 tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc
     1381 cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta
     1441 gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta
     1501 aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg
     1561 gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga
     1621 atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg
     1681 tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa
     1741 agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga
     1801 gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct
     1861 tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt
     1921 tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact
     1981 gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt
     2041 gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt
     2101 tttcttccat ttcaggtgtc gtgaggtacc ggatcctcta gagtcgactc cggaataact
     2161 tcgtatagga tactttatac gaagttatgc agaatggtag ctggattgta gctgctatta
     2221 gcaatatgaa acctcttaat aacttcgtat agcatacatt atacgaagtt atggcgcgcc
     2281 ttaagcgtaa tctggaacat cgtatgggta aaactccccg gacccagctg ctaaatggct
     2341 ctgatacaag ctgtggtaat cctctttctc catgaggagt ggctgctttt cttcaggaag
     2401 ctgctttctc ttgtgacgac acagcaagct cacaagccct gccagcaggg cagtgaggac
     2461 ggcccctacc atcgccgccc caaggagcca tgaccagatc cgactcgctt gttccaaata
     2521 ggacttaatg tagtcttgaa aagagtctgg gtctgaatct tgtagatagc tatagtcata
     2581 gcccagatct ttggatgaaa taaagaaatc accatttctg tacagtggta taaaaggaac
     2641 catgtaggat tcccggttat gtccaatggg tgcattggct tctggataaa cttcttgaag
     2701 aggacggtgc cttcggagcc actgctcaaa aatactgtca acaaatgcat ggtgaagaag
     2761 gaagatagga tcgttggcag atccctgtac ctgggacatt gttccattca tatagatgtg
     2821 caaggcattg tgcatgctgc tttgagaggc atccgctatc ccagtaagtg gactagcaaa
     2881 tccttccagt gtatttctaa agctgaaatt ggcagcttta tccatggaac cagattcata
     2941 ttgggtcaaa ctcaggcaaa attctacatc agctgaagag gggagccttg gggttctgga
     3001 tttgtcatgg tttccaggat tacgccgtaa aggtccctcg ggcgttccat tgcataaaga
     3061 ctgatggctg ttgtactcct ccaatcggct acagacaatc tgccaagagg agaagaatga
     3121 tgctgggctg agtaagttag gatttgtggg gtgctgacct cccatgtact catctgtgca
     3181 aatgtcacac ttttctgcat cccgccagtc ccaatatgga atagtgaagt tttcatctcc
     3241 tgtcagcttc tggatttctt gttcccaccg caacaagaag agtctatgcc aaggcagaaa
     3301 agctggtgct tcatgggcaa aatcaatgtc tctccagatt tcagatcccc caagcagtgc
     3361 atccattgac acataataat gcatccagac aaagaggtca taaatattga tgtcgttaaa
     3421 catgggtgtt gatccatttt tcatttggcc ataggtccct atggggatga catagtctga
     3481 gctgatggta tgctttgcta aagtgaggta ggcaaaaaat ttgtccttct ctggggcact
     3541 caaatcgaag atgtttcttc tcaccaagag tcgtctctct gtgcagtttg gtccccaaaa
     3601 gccaaacttg cagtttccac agttgaatcc catgaagttg ccagagcact ggcaggtcct
     3661 attataaaag acggaaggcc acgactcccg gtcatccacc cctgtgaagg gaaattgagg
     3721 cccaagtggt gcattggaca gaaggatatt ctgacaggaa cctctgcctg aaagctggcc
     3781 acagggactc ctgtccccgc tccacggtgg acagcattcc ttctccatca ggttcttaga
     3841 ggagacacag gctctaggga aatggccagc ggaggtctgg aaactccaca gcaggcagta
     3901 caaaacagcc aggagcatca aggtcttagg ctagcataac ttcgtataaa gtatcctata
     3961 cgaagttatt tgccttaacc cagaaattat cactgttatt ctttagaatg gtgcaaagaa
     4021 taacttcgta taatgtatgc tatacgaagt tatgaattca agcttaatca acctctggat
     4081 tacaaaattt gtgaaagatt gactgatatt cttaactatg ttgctccttt tacgctgtgt
     4141 ggatatgctg ctttaatgcc tctgtatcat gctattgctt cccgtatggc tttcgttttc
     4201 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtccgt
     4261 caacgtggcg tggtgtgctc tgtgtttgct gacgcaaccc ccactggctg gggcattgcc
     4321 accacctgtc aactcctttc tgggactttc gctttccccc tcccgatcgc cacggcagaa
     4381 ctcatcgccg cctgccttgc ccgctgctgg acaggggcta ggttgctggg cactgataat
     4441 tccgtggtgt tgtcggggaa gctgacgtca gcgctaataa aatcgctatc catcgaagat
     4501 ggatgtgtgt tggttttttg tgtgggtaac ccacgtgcgg accgagcggc cgcaggaacc
     4561 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     4621 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     4681 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4741 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     4801 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     4861 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4921 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4981 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5041 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5101 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     5161 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     5221 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     5281 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     5341 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     5401 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5461 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5521 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5581 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5641 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5701 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5761 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5821 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5881 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5941 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6001 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6061 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6121 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6181 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6241 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6301 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6361 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6421 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6481 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6541 taggtgaaga tcctttttga taatctcatg accaaa
//
