LOCUS       sequence-496975-        7236 bp    DNA     circular SYN 12-JAN-2026
DEFINITION  Control hTyr (D383N/A206T) — human tyrosinase carrying D383N and 
            A206T substitutions; catalytic activity super slow; intended as an 
            enzymatically-impaired control..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7236)
  TITLE     Induces NM-like pigment accumulation in catecholaminergic neurons
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7236)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 12, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAV_Ef1a_DIO_hTyrHA_superslowControl
FEATURES             Location/Qualifiers
     source          1..7236
                     /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        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1440..1473)
                     /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(1524..1557)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1569..1595)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    3221..3254
                     /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    3305..3338
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3363..3951
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3416..3436)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3834..3845)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3983..4459
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4182..4201)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4499..4639
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4499..4628
                     /label=AAV2 ITR
     rep_origin      4714..5169
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4801..4820)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5011..5032
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5186..5205)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5305..5327
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5365..5383)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5451..5555
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5556..5624,5625..6416)
                     /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(5774..5793)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6587..7175
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7076..7095
                     /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 gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa
     1441 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
     1501 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
     1561 gcgccttaag cgtaatctgg aacatcgtat gggtaaaact ccccggaccc agctgctaaa
     1621 tggctctgat acaagctgtg gtaatcctct ttctccatga ggagtggctg cttttcttca
     1681 ggaagctgct ttctcttgtg acgacacagc aagctcacaa gccctgccag cagggcagtg
     1741 aggacggccc ctaccatcgc cgccccaagg agccatgacc agatccgact cgcttgttcc
     1801 aaataggact taatgtagtc ttgaaaagag tctgggtctg aatcttgtag atagctatag
     1861 tcatagccca gatctttgga tgaaataaag aaatcaccat ttctgtacag tggtataaaa
     1921 ggaaccatgt aggattcccg gttatgtcca atgggtgcat tggcttctgg ataaacttct
     1981 tgaagaggac ggtgccttcg gagccactgc tcaaaaatac tgtcaacaaa tgcatggtga
     2041 agaaggaaga taggattgtt ggcagatccc tgtacctggg acattgttcc attcatatag
     2101 atgtgcaagg cattgtgcat gctgctttga gaggcatccg ctatcccagt aagtggacta
     2161 gcaaatcctt ccagtgtatt tctaaagctg aaattggcag ctttatccat ggaaccagat
     2221 tcatattggg tcaaactcag gcaaaattct acatcagctg aagaggggag ccttggggtt
     2281 ctggatttgt catggtttcc aggattacgc cgtaaaggtc cctcgggcgt tccattgcat
     2341 aaagactgat ggctgttgta ctcctccaat cggctacaga caatctgcca agaggagaag
     2401 aatgatgctg ggctgagtaa gttaggattt gtggggtgct gacctcccat gtactcatct
     2461 gtgcaaatgt cacacttttc tgcatcccgc cagtcccaat atggaatagt gaagttttca
     2521 tctcctgtca gcttctggat ttcttgttcc caccgcaaca agaagagtct atgccaaggc
     2581 agaaaagttg gtgcttcatg ggcaaaatca atgtctctcc agatttcaga tcccccaagc
     2641 agtgcatcca ttgacacata ataatgcatc cagacaaaga ggtcataaat attgatgtcg
     2701 ttaaacatgg gtgttgatcc atttttcatt tggccatagg tccctatggg gatgacatag
     2761 tctgagctga tggtatgctt tgctaaagtg aggtaggcaa aaaatttgtc cttctctggg
     2821 gcactcaaat cgaagatgtt tcttctcacc aagagtcgtc tctctgtgca gtttggtccc
     2881 caaaagccaa acttgcagtt tccacagttg aatcccatga agttgccaga gcactggcag
     2941 gtcctattat aaaagacgga aggccacgac tcccggtcat ccacccctgt gaagggaaat
     3001 tgaggcccaa gtggtgcatt ggacagaagg atattctgac aggaacctct gcctgaaagc
     3061 tggccacagg gactcctgtc cccgctccac ggtggacagc attccttctc catcaggttc
     3121 ttagaggaga cacaggctct agggaaatgg ccagcggagg tctggaaact ccacagcagg
     3181 cagtacaaaa cagccaggag catcaaggtc ttaggctagc ataacttcgt ataaagtatc
     3241 ctatacgaag ttatttgcct taacccagaa attatcactg ttattcttta gaatggtgca
     3301 aagaataact tcgtataatg tatgctatac gaagttatga attcgatatc aagcttatcg
     3361 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     3421 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     3481 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     3541 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     3601 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     3661 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     3721 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     3781 tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     3841 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     3901 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc
     3961 gagcgctgct cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct
     4021 ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat
     4081 cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag
     4141 caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg
     4201 gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct
     4261 cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt
     4321 tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta
     4381 atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac
     4441 tgctcccttc cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag
     4501 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     4561 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     4621 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     4681 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     4741 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     4801 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4861 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4921 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4981 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     5041 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
     5101 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     5161 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     5221 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     5281 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     5341 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     5401 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     5461 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     5521 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     5581 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     5641 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     5701 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     5761 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     5821 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     5881 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     5941 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     6001 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     6061 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     6121 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     6181 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     6241 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     6301 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     6361 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     6421 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     6481 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     6541 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6601 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6661 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6721 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6781 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6841 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6901 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6961 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     7021 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     7081 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     7141 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     7201 ggttcctggc cttttgctgg ccttttgctc acatgt
//
